Article

Fibroblast Growth Factor 5 Inhibits Hair Growth by Blocking Dermal Papilla Cell Activation

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Abstract

Fibroblast growth factor (FGF) 5 inhibits hair growth and induces catagen in mouse hair follicles, in vivo. Given that FGF-5 receptor (FGFR1) is expressed in dermal papilla cells (DPCs), which are known to stimulate outer root sheath cell (ORSC) proliferation, we hypothesized that FGF-5 attenuates DPC-mediated ORSC proliferation. In the present study, DPCs and ORSCs were isolated from rat vibrissae, after which the effects of FGF-5 on proliferation of ORSCs cultured in DPC-conditioned medium were assessed. We first confirmed that FGFR1 was expressed in cultured DPCs and detected FGFR2-4 as well. ORSC proliferation was increased approximately twofold when the cells were cultured in DPC-conditioned medium, and the effect was unaltered by FGF-5. In addition, FGF-5 did not directly inhibit ORSC proliferation; indeed, it actually promoted proliferation of both DPCs and ORSCs. When DPCs were first activated by exposure to FGF-1 and FGF-2, which are expressed in hair follicles during anagen, ORSC proliferation observed in the resultant conditioned medium was substantially greater than in medium conditioned by unstimulated DPCs. The FGF-1-induced enhancement was reversed by FGF-5, diminishing ORSC proliferation to control levels. By contrast, the enhancement of DPC-mediated ORSC proliferation by FGF-2 was not suppressed by FGF-5. Proliferation of ORSCs did not depend on DPC proliferation, nor did FGF-1 directly promote ORSC proliferation. Dermal papillae thus appear to require activation before they will efficiently stimulate hair growth, and FGF-5 appears to inhibit hair growth and induce catagen by blocking that activation.

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... A study from Higgins et al. (2014) indicates that human hair follicle, cultured in vitro, enters into catagen phase prematurely when exposed to recombinant FGF5. Moreover, the administration of FGF5 inhibits mouse hair growth by blocking the activation of dermal papilla cells (Ota et al., 2002). These previous studies suggest that FGF5 may function as a negative regulator in hair growth. ...
... In accordance with this previous finding, our group has prior identified two products (FGF5 and FGF5s) generated from FGF5 gene in cashmere goat (He et al., 2014). Ota et al. (2002) have proved that FGF5 inhibited hair growth and induced catagen by blocking dermal papilla cell activation in mice. FGF signal transduction encompasses ligand binding to a high-affinity receptor-tyrosine kinase (Rosenquist and Martin, 1996). ...
... FGF signal transduction encompasses ligand binding to a high-affinity receptor-tyrosine kinase (Rosenquist and Martin, 1996). FGFR1 is a high-affinity receptor to FGF5 (Ornitz and Leder, 1992), and has been demonstrated to be exclusively expressed in dermal papilla within HF (Rosenquist and Martin, 1996;Suzuki et al., 1998;Ota et al., 2002). Cashmere goat has a double-coat; we therefore first isolated dermal papilla cells from PHF and SHF, and found that FGFR1 was expressed in both types of cashmere goat dermal papilla cells. ...
Article
To determine the relationship between fibroblast growth factor 5 (FGF5) and FGF5-short (FGF5s) in dermal papilla cells of cashmere goat primary and secondary hair follicles. We isolated dermal papilla cells from primary hair follicle (PHF) and secondary hair follicle (SHF) of cashmere goat, and found that the FGF5 receptor, fibroblast growth factor receptor 1 (FGFR1), was expressed in these two types of dermal papilla cells. Moreover, adenovirus-mediated overexpression of FGF5 could upregulate the mRNA expression of insulin-like growth factor-1 (IGF-1), versican and noggin that were important for follicle growth maintenance, whereas downregulate the expression of anagen chalone bone morphogenetic protein 4 (BMP4) in dermal papilla cells. However, these alterations were partly reversed by FGF5s overexpression. In conclusion, our results demonstrated that FGF5s acted as an inhibitor of FGF5 in the regulation of anagen-catagen transition of cashmere goat dermal papilla cells.
... Intriguingly, each cell subset exhibits distinctive FGF expression profile. Human FGFs are largely divided into 7 subgroups based on their biochemical characteristics, namely, FGF1 (1, 2), FGF4 (4, 5, 6), FGF7 (3, 7, 10, 12), FGF8 (8,7,18), FGF9 (9,16,20), FGF11 (11,12,13,14), and FGF15/19 (19,21,23) subfamilies [39]. The representative results of RT-PCR analyses are presented in Fig. 1. ...
... FGF expression profiling in HF-related dermal cell subsets; DPCs, DSCs, and sFBs first suggested a unique FGF expression pattern in each subgroup. In an examined sample, sFBs were characterized by higher expression of FGF5 and FGF18, which individually induce a catagen [20][21][22][23] or a telogen [34][35][36] in animal models, compared to those in DPCs and DSCs. FGF13 expression was also relatively strong in sFBs. ...
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Background: Hair follicle (HF) formation and growth are sustained by epithelial-mesenchymal interaction via growth factors and cytokines. Pivotal roles of FGFs on HF regeneration and neogenesis have been reported mainly in rodent models. FGF expression is regulated by upstream pathways, represented by canonical WNT signaling; however, how FGFs influence on human folliculogenesis remains elusive. The aim of this study is to assess if human scalp-derived fibroblasts (sFBs) are able to modulate their FGF expression profile in response to WNT activation and to evaluate the influence of WNT-activated or suppressed FGFs on folliculogenesis. Methods: Dermal papilla cells (DPCs), dermal sheath cells (DSCs), and sFBs were isolated from the human scalp and cultured independently. The gene expression profile of FGFs in DPCs, DSCs, and sFBs and the influence of WNT activator, CHIR99021, on FGF expression pattern in sFBs were evaluated by reverse transcription polymerase chain reaction, which were confirmed at protein level by western blotting analysis. The changes in the expression of DPC or keratinocyte (KC) biomarkers under the presence of FGF7 or 9 were examined in both single and co-culture assay of DPCs and/or KCs. The influence of FGF 7 and FGF 9 on hair morphogenesis and growth was analyzed in vivo using mouse chamber assay. Results: In single culture, sFBs were distinguished from DPCs and DSCs by relatively high expression of FGF5 and FGF18, potential inducers of hair cycle retardation or catagen phase. In WNT-activated state, sFBs downregulated FGF7 while upregulating FGF9, a positive regulator of HF morphogenesis, FGF16 and FGF20 belonging to the same FGF subfamily. In addition, CHIR99021, a WNT activator, dose-dependently modulated FGF7 and 9 expression to be folliculogenic. Altered expressions of FGF7 and FGF9 by CHIR99021 were confirmed at protein level. Supplementation of FGF9 to cultured DPCs resulted in upregulation of representative DP biomarkers and this tendency was sustained, when DPCs were co-cultured with KCs. In mouse chamber assay, FGF9 increased both the number and the diameter of newly formed HFs, while FGF7 decreased HF diameter. Conclusion: The results implied that sFBs support HF formation by modulating regional FGF expression profile responding to WNT activation.
... During the anagen phase, hair is thought to grow as a result of the proliferation of outer root sheath cells (ORSCs) that are induced by humoral factors that are synthesized and released by dermal papilla cells (DPCs). Further in vitro results have demonstrated that FGF5 has a role in inhibiting hair growth and inducing catagen by blocking the activation of DPCs [7,17,18]. After the blocking effect, it appears that FGF5 regulates the anagen to catagen transition by interacting with members of other growth factor families (BMP, TGF-β, EGF, VERSICAN, WNT, NOTCH, and SHH) [18][19][20][21][22]. ...
... The secretion of FGF5 interacts with the receptor FGFR1, which is expressed in dermal papilla cells (DPCs); this signal is known to stimulate outer root sheath cell (ORSC) proliferation [17]. Other molecular factors were also found to play a role in the hair cycle. ...
Article
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Hair growth and morphology are generally regulated by the hair cycle in mammals. Fibroblast Growth Factor 5 (FGF5), which is a hair cycle regulator, has a role in regulating the hair cycle during the transition from the anagen phase to the catagen phase, and a hereditary long hair phenotype has been widely reported when FGF5 is mutated in humans and other species. However, there has been no such report in rabbits. Thus, the first exon of rabbit FGF5 was disrupted by the CRISPR/Cas9 system, and the phenotype of FGF5-/- rabbits was characterized while using hematoxylin and eosin (H&E) staining, immunohistochemistry, quantitative PCR, scanning electron microscopy, and western blotting. The results showed a significant and systemic long hair phenotype in the FGF5-/- rabbits, which indicated that FGF5 is a negative regulator of hair growth. In addition, a decreased diameter of the fiber and a higher area proportion of hair follicle clusters were determined in FGF5-/- rabbits as compared with the WT rabbits. Further investigation verified that prolonging the anagen phase in rabbits, with decreased BMP2/4 pathway signaling and increased VERSICAN pathway signaling, caused the systemic long hair phenotype. Taken together, these results indicate a systemic long hair phenotype by prolonging anagen in FGF5-/- rabbits, which could be widely used for Fur production and an ideal model for studying the mechanism of long hair in the future.
... The most important role of the dermal papilla is the regulation of hair growth. Signals to regulate hair growth from the dermal papilla stimulate hair follicle cells (such as outer root sheath cells), and then hair growth or hair regression is promoted [3]. The most useful and rapid method for evaluating hair growth promotion is by determining the dermal papilla growth rate as influenced by various samples [4]. ...
... The most useful and rapid method for evaluating hair growth promotion is by determining the dermal papilla growth rate as influenced by various samples [4]. After that, more detailed studies should be performed such as in vivo assay34567891011. We collected many aromatic plants from Tunisia, which is located in North Africa. ...
Chapter
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Recently, people are experiencing more stress and some have problems regarding hair growth as a part of the aging process. Resolving this hair growth problem is important for the aging society in the future. This is because not only bodily health but also mental health is important for a healthy life. Thus, we attempted to find plants having hair growth regulation activity. The Republic of Tunisia is located in North Africa; its northern boundary faces the Mediterranean Sea while its southern side leads to the Sahara Desert. The distance during the Mediterranean Sea and the Sahara Desert is only few kilometers. This environment can be considered that dry inclination is very high. In such environment, plants can accumulate antistress factors in their system. Thus, we collected many plant extracts from Tunisia for bioprospecting purposes. We show that a Tunisian aromatic plant extract has a high activity for promotion of hair growth cycle, or induction of anagen phase from telogen phase. KeywordsHair growth cycle-dermal papilla-MTT assay-vasodilatation
... However, they were clearly detected in T-cell population isolated from the skin of patients with psoriasis [13]. This is consistent with the previously suggested role of IL-22 in psoriasis [74,75]. Firstly, the Th22 expression profile secretes several fibroblast growth factors (FGFs). ...
... Firstly, the Th22 expression profile secretes several fibroblast growth factors (FGFs). FGF1 is a powerful mitogen exhibiting strong action in different cells types, including endothelial cells [76,77], while FGF5 acts on neural differentiation [78] and is associated with inhibition of hair growth [74,75]. These cytokines play a role in various stages of development as well as in angiogenesis and wound healing processes. ...
Article
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Introduction: Th22 and related cytokines regulate various processes and have been linked to diverse effects. The levels of Th22 and cytokine IL-22 are increased in several disorders and positively related to some autoimmune diseases. Preclinical studies have suggested that the inhibition or stimulation of IL-22 is an attractive approach to reverse the immune disorders. Simultaneously, considering many patients with refractory autoimmune diseases respond poorly to the therapies which are highly toxic, more effective therapies are to be examined. Areas covered: The role of Th22 cells and related cytokines and therapeutic strategies targeting them in many illnesses, especially inflammatory and autoimmune diseases. Expert opinion: Th22 cells and related cytokine IL-22 regulate multiple biological functions and play an important role in a number of inflammatory and autoimmune diseases. They have unique and attractive advantages for targeting. Targeting IL-22 has already been trialed in many mice experiments, showing better efficacy and fewer side effects compared with classical drugs. Targeting IL-22 or Th22 might provide pathogenetic treatment. However, Th22 subset is a recently identified Th subset and its associated research is extremely limited. Therefore, there are still many unanswered questions and further researches are warranted.
... Increasing number of studies indicate that the FGF5 gene is one of modulators of hair cycle in mice. FGF5 inhibits the hair growth and induces catagen, whereas FGF5s antagonizes the activity of FGF5 during anagen (Hattori et al., 1996;Suzuki et al., 1998Suzuki et al., , 2000Ota et al., 2002). ...
... A series of studies in mice hair follicle have indicated that FGF5 can inhibit hair growth by blocking dermal papilla cell activation and the two FGF5 gene products regulated the hair cycle: FGF5 promotes the transition from anagen to catagen, whereas FGF5s antagonizes such activity during anagen (Hébert et al., 1994;Suzuki et al., 2000;Ota et al., 2002). Moreover, Suzuki et al. (1998) suggested that the function of FGF5 is more likely in the induction of catagen than in the inhibition of hair growth and that FGF5s modifies the function of FGF5. ...
... Since the efficiency of the third-generation base editor (BE3; rAPOBEC1-nCas9-UGI) was shown to be better than BE1 (rAPOBEC1-dCas9) and BE2 (rAPO-BEC1-dCas9-UGI) [7,12], BE3 was selected to induce nonsense mutations into the coding sequence of the caprine target gene, fibroblast growth factor 5 (FGF5). The encoded protein of FGF5 is secreted during the hair growth cycle to signal inhibition of hair growth by a mechanism that blocks dermal papilla cell activation [19]. FGF5 is regarded as the causative gene responsible for the angora phenotype (long hair) in mice [20], and we have previously shown that disruption of FGF5 via CRISPR/Cas9 resulted in longer hair fibers and a 30% increase in cashmere yield per animal [21,22]. ...
... Given that FGF5 was known to inhibit hair growth by blocking dermal papilla cell activation [19], and was a [25], cats [26], mice [20], and humans [27], characterization of fibers in the founder animals would reveal the degree of penetrance of the mosaic genotypes on phenotypes. First, the length of hair fibers (outer coat hair and inner fine fiber) between mutant and control animals was measured and compared. ...
Article
The ability to alter single bases without homology directed repair (HDR) of double‐strand breaks provides a potential solution for editing livestock genomes for economic traits, which are often multigenic. Progress towards multiplex editing in large animals has been hampered by the costly inefficiencies of HDR via microinjection of in vitro manipulated embryos. Here, we designed sgRNAs to induce nonsense codons (C‐to‐T transitions) at four target sites in caprine FGF5, which is a crucial regulator of hair length in mammals. Initial transfections of the third generation Base Editor (BE3) plasmid and four different sgRNAs into caprine fibroblasts were ineffective in altering FGF5. In contrast, all five progenies produced from microinjected single‐cell embryos had alleles with a targeted nonsense mutation. The effectiveness of BE3 to make single base changes varied considerably based on sgRNA design. In addition, the rate of mosaicism differed between animals, target sites, and tissue type. The phenotypic effects on hair fiber were characterized by hematoxylin and eosin (H&E), immunofluorescence staining, and western blotting. Differences in morphology were detectable, even though mosaicism was probably affecting the levels of FGF5 expression. PCR amplicon and whole genome resequencing analyses for off‐target changes caused by BE3 were low at a genome‐wide scale. This study provided the first evidence of base editing in large mammals produced from microinjected single cell embryos. Our results support further optimization of BEs for introgressing complex human disease alleles into large animal models, to evaluate potential genetic improvement of complex health and production traits in a single generation. This article is protected by copyright. All rights reserved.
... FGF1 + Involved in HF differentiation, prevents radiation-induced apoptosis in HF [21][22][23] FGF2 + Involved in the proliferation of HF cells, elongation of the anagen phase [24][25][26] FGF5 − Catagen induction, blocks DPC activation during anagen [27][28][29] FGF7 + Elongation of the anagen phase, hair germ proliferation, stem cell activation [3,30] FGF8 − Inhibits proliferation of epidermal cells [ [41] FGF12 lacks the N-terminal signal sequence, unlike most FGF family members. Instead, it contains basic residue clusters which allow FGF12 to have nuclear localization signal. ...
Article
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The fibroblast growth factor (FGF) family has various biological functions, including cell growth, tissue regeneration, embryonic development, metabolism, and angiogenesis. In the case of hair growth, several members of the FGF family, such as FGF1 and FGF2, are involved in hair growth, while FGF5 has the opposite effect. In this study, the regulation of the hair growth cycle by FGF12 was investigated. To observe its effect, the expression of FGF12 was downregulated in mice and outer root sheath (ORS) by siRNA transfection, while FGF12 overexpression was carried out using FGF12 adenovirus. For the results, FGF12 was primarily expressed in ORS cells with a high expression during the anagen phase of hair follicles. Knockdown of FGF12 delayed telogen-to-anagen transition in mice and decreased the hair length in vibrissae hair follicles. It also inhibited the proliferation and migration of ORS cells. On the contrary, FGF12 overexpression increased the migration of ORS cells. FGF12-overexpressed ORS cells induced the telogen-to-anagen transition in the animal model. In addition, FGF12 overexpression regulated the expression of PDGF-CC, MDK, and HB-EGF, and treatment of these factors exhibited hair growth promotion. Altogether, FGF12 promoted hair growth by inducing the anagen phase of hair follicles, suggesting the potential for hair loss therapy.
... FGF5 as a regulator of hair growth has been proved to be responsible for long hair phenotype by targeted and spontaneous loss-of-function mutations. It inhibits the activation of dermal papilla cells proliferation and synthesis of hair fiber [22]. Wide type FGF5 protein owns critical βstand motif involved in interaction between FGF5 and its receptor to trigger receptor activation and biological responses [23]. ...
Article
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Fibroblast growth factor 5 (FGF5) regulates hair length in humans and a variety of other animals. To investigate whether FGF5 has similar effects in sheep, we used CRISPR/Cas9 to generate loss-of-function mutations with the FGF5 gene in Chinese Merino sheep. A total of 16 lambs were identified with genetic mutations within the targeting locus: 13 lambs had biallelic modifications and 3 lambs had monoallelic modifications. Characterization of the modifications revealed that 13 were frame shift mutations that led to premature termination; the other 3 were in-frame deletions. Thus, CRISPR/Cas9 efficiently generated loss-of-function mutations in the sheep FGF5 gene. We then investigated the effect of loss of FGF5 function on wool traits in 12 lambs, and found that the wool staple length and stretched length of genetically modified yearling sheep were significantly longer than that of wild-type control animals. The greasy fleece weight of genetically modified yearling sheep was also significantly greater than that of wild-type sheep. Moreover, the mean fiber diameter in the genetically modified versus wild type sheep showed no significant difference, suggesting that the increase of greasy fleece weight was likely attributed to the increase in wool length. Our work suggests that CRISPR/Cas9-mediated loss of FGF5 activity could promote wool growth, and consequently increase wool length and yield. This article is protected by copyright. All rights reserved.
... The DEPICT analysis (Supplementary Tables 4-6) and literature search identified highly plausible candidate genes, such as FGF5 at 4q21.21 (rs982804; P ¼ 2.2 Â 10 À 9 , METAL) and DKK2 at 4q25 (rs145945174; P ¼ 1.3 Â 10 À 13 , METAL). FGF5 plays an important role in the regulation of anagen-to-catagen transition and the control of human hair length [20][21][22] . DKK2 encodes for a member of the family of dickkopf WNT-signalling inhibitors, which are reported to be secreted by dermal papilla cells (DPCs) ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms14694 in response to androgens and to promote androgen-induced (premature) anagen-to-catagen transition 23,24 . ...
Article
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Male-pattern baldness (MPB) is a common and highly heritable trait characterized by androgen-dependent, progressive hair loss from the scalp. Here, we carry out the largest GWAS meta-analysis of MPB to date, comprising 10,846 early-onset cases and 11,672 controls from eight independent cohorts. We identify 63 MPB-associated loci (P<5 × 10−8, METAL) of which 23 have not been reported previously. The 63 loci explain ∼39% of the phenotypic variance in MPB and highlight several plausible candidate genes (FGF5, IRF4, DKK2) and pathways (melatonin signalling, adipogenesis) that are likely to be implicated in the key-pathophysiological features of MPB and may represent promising targets for the development of novel therapeutic options. The data provide molecular evidence that rather than being an isolated trait, MPB shares a substantial biological basis with numerous other human phenotypes and may deserve evaluation as an early prognostic marker, for example, for prostate cancer, sudden cardiac arrest and neurodegenerative disorders.
... Studies in rodents have found that FGF5 messenger RNA (mRNA) is upregulated during late anagen phase, 20 DP cells that express the receptor for FGF5 (FGFR1) 21 are responsive to FGF5 in vitro 22 and FGF5 inhibits DP cell activation. 23 Such findings demonstrate that FGF5 signaling to DP cells through FGFR1 is a critical regulatory process. Furthermore, FGF5 accumulates in the outer root sheath and in small macrophage-like cells surrounding the follicle, and the expression at these sites is increased at late anagen phase. ...
Article
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Background There are very few effective, scientifically validated treatments with known mechanisms of action for treatment of hair loss in both men and women. Fibroblast growth factor 5 (FGF5) is an important factor in the irreversible transition from anagen to catagen, and inhibition of FGF5 prolongs anagen phase and reduces hair loss. Objective We aimed to screen botanically derived molecules for FGF5 inhibitory activity in vitro and assess efficacy in a clinical setting. Methods We screened for FGF5 inhibitory efficacy via a novel 2-step in vitro pipeline consisting of an engineered FGF5 responsive cell line, followed by an activated dermal papillae (DP) cell method. Efficacy in a clinical setting was assessed in a randomized, single-blind, placebo-controlled trial against early- to mid-stage pattern hair loss in men and women. Results We observed FGF5 inhibitory activity for a number of compounds from the monoterpenoid family, many showing greater inhibitory efficacy than our previously reported crude plant extracts. Evaluation of a lead candidate in a clinical study over 112 days showed a significant improvement in anagen:telogen (AT) ratio (p = 0.002), reduced hair fall (p = 0.007) and improved visual grading (p = 0.004). Scientifically matched photography on a subgroup of randomly chosen participants highlighted significant improvement in hair density, with increases evident in all tested participants compared to baseline. Conclusion Isolates from the monoterpenoid family displayed efficacy in FGF5 inhibition in vitro. A topical formulation containing a leading isolate significantly improved AT ratio, reduced hair fall and increased apparent hair density in the tested population of men and women.
... Signaling factors such as Wnt and sonic hedgehog 28 , growth factors, as well as transmembrane and extracellular matrix molecules, seem to be implicated in that process initiated by the androgens 29 . HS and CS modulate the actions of growth factors such as fibroblast growth factor (FGF) 30,31 , vascular endothelial growthfactor (VEGF) 32 , bone morphogenetic proteins (BMPs) 33 , Wnt 28 and sonic hedgehog 28 , which are known to be involved in the biology of the hair follicle 12,34 . It is known that dermal papilla is rich in GAGs but as shown in patients with an-drogenetic alopecia (AGA) 35 , androgens activity is related to GAGs reduction. ...
Article
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Background There is evidence that glycosaminoglycans (GAGs) are present in the hair shaft within the follicle but there are no studies regarding GAGs isolation and measurement in the human hair shaft over the scalp surface, it means, in the free hair shaft. Objective The purpose of our research was to isolate and measure the total GAGs from human free hair shaft. Methods Seventy-five healthy individuals participated in the study, 58 adults, men and women over the age of 50 and 17 children (aged 4~9). GAGs in hair samples, received from the parietal and the occipital areas, were isolated with 4 M guanidine HCl and measured by the uronic acid-carbazole reaction assay. Results GAGs concentration was significantly higher in the occipital area than in the parietal area, in all study groups. GAG levels from both areas were significantly higher in children than in adults. GAG levels were not associated with gender, hair color or type. Conclusion We report the presence of GAGs in the human free hair shaft and the correlation of hair GAG levels with the scalp area and participants' age.
... Este factor, normalmente se expresa en la ORS en el proceso de inducción de catagen y bloquea la activación de la papila dérmica (Ota et al., 2002). ...
Thesis
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Mohair and cashmere production and quality traits are important characteristics in the selection and conservation programmes in Angora and Criollo Neuquino (CN) goats. These characteristics are affected by hair follicle groups in the goat skin. The utilization of DNA information associated to these characteristics would lead to obtain sensitive information about their genetic determinism. The goal of this study was to detect Quantitative Trait Loci (QTL) affecting hair follicle goat´s skin, fibre production and quality characteristics on chromosome 1 (CHI1), CHI2, CHI5, CHI13 and CHI19. The experimental design involved seven paternal half-sib families in an Angora x CN backcross (BC) population. Two BC Angora and five BC CN families totalized 354 offspring used for genetic mapping. A total of 43 microsatellites (MS) were used in the following way: 9 of them were distributed in 186 cM on CHI1, 9 in 127 cM studied on CHI2 and 12 were distributed on CHI5. Likewise, 9 MS were distributed to cover 36 cM on CHI13 and 9 were used to study 69 cM on CHI19. During 3 years 24 phenotypic characteristics of skin and fibre were recorded. These traits were corrected by six factors: sex, year, litter size, moment of birth at season, age of dam at birth, and cross type using the MIXED procedure of the SAS software package. With the appropriate combination of significant (p<0.10) fixed effects for each trait, contemporary groups were constructed to be used in the linkage analysis (LA). The LA was made using multiple regression intervals mapping with the web version of GridQTL software. Sixteen QTL were detected. Three QTL affected the secondary to primary follicle ratio on CHI1 and CHI19. Number of secondary follicles within follicle group on CHI19, staple length on CHI2 and CHI19. Cashmere yield were affected by CHI13 and CHI19; Average fibre diameter was affected by CHI13 and CHI5 and washing yield was affected by CHI2. Finally, clean and greasy fleece weight, mean cashmere diameter and percentage of fibre greater than 30 μm were all affected by CHI13. These results confirmed some previous reports and opened the door to in-depth studies of candidate genes analysis included in the chromosomal regions. There have been demonstrated, for the first time in mammals, QTL affecting quantitative traits related to the hair follicles in the skin. The cited chromosome regions should be subject of future fine mapping analysis, with the ultimate aim of finding allelic variants of candidate genes responsible for those phenotypic variations.
... Furthermore, lncRNA2919 could dysregulate HF cycle-and growth-related genes. It upregulated the expression of FGF5 [29,30], KRTAP11-1 [31], and STAT1 [32], whereas it downregulated the expression of LEF1 [33], WNT2 [34], BCL2 [35,36], and CCND1 [37]. ...
Article
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Hair follicles (HFs) are organs that periodically regenerate during the growth and development of mammals. Long non-coding RNAs (lncRNAs) are non-coding RNAs with crucial roles in many biological processes. Our previous study identified that lncRNA2919 is highly expressed in catagen during the HF cycle. In this study, the in vivo rabbit model was established using intradermal injection of adenovirus-mediated lncRNA2919. The results showed that lncRNA2919 decreased HF depth and density and contributed to HF regrowth, thereby indicating that lncRNA2919 plays a negative role in HF regeneration. Moreover, methylation levels of the lncRNA2919 promoter at different HF cycle stages were detected through bisulfite sequencing. The key CpG site that negatively correlates with lncRNA2919 expression during the HF cycle was identified. 5-Aza-dc-induced demethylation upregulated lncRNA2919 expression, and the core promoter region of lncRNA2919 was verified on the basis of luciferase activity. Furthermore, we found that DNA methylation could prevent the binding of EGR1 to the lncRNA2919 promoter region, thereby affecting the transcriptional expression of lncRNA2919. Collectively, DNA methylation inhibits the transcriptional expression of lncRNA2919, which plays a vital role in the HF cycle and HF regrowth. These findings contribute to the basic theory of epigenetics in HF biology and provide references for further research in HF disease treatment and animal wool production.
... In mouse, FGF5 mRNA is highly expressed in the hair follicle as two isoforms, identified as FGF5 and FGF5S (Suzuki et al., 2000), and the latter is due to the alternative splicing of exon 2 (Hattori et al., 1996). Both isoforms, through binding to FGF receptor 1 and 2, regulate the hair follicle growth cycle during the anagen stage: FGF5 actively inhibits cell proliferation and the synthesis of hair fibers, while FGF5S antagonizes the inhibitory effects of FGF5 through competitively binding the FGF receptors (Suzuki et al., 2000;Ota et al., 2002;He et al., 2016). ...
Article
Article freely available (untill March 01, 2018) at https://authors.elsevier.com/a/1WMwK1L~GAarkW Two different phenotypes are described in alpaca, identified as suri and huacaya, which differ in the type of fleece. The huacaya fleece is characterized by compact, soft and highly crimped fibers, while the suri fleece is longer, straight, less-crimped and lustrous. In our study, the Fibroblast growth factor 5 (FGF5) was investigated as a possible candidate gene for hair length in alpaca (Vicugna pacos). As previously identified in other mammals, our results show that the alpaca FGF5 gene gives rise to a short (FGF5S) and a long (FGF5) isoform. Interestingly, in the long isoform, we observed a point mutation (i.e., a transition C>T at position 499 downstream of the ATG codon) that is able to generate a premature termination codon (PTC). The highly conserved nucleotide and amino acid sequence after PTC suggested a readthrough event (RT) that was confirmed by western blot analysis. The analysis of cDNA sequence revealed motifs and structures of mRNA undergoing RT. In fact, the event is positively influenced by particular signals harbored by the transcript. To the best of our knowledge, this is the first case of a readthrough event on PTC reported for the FGF5 gene and the first case of this translational mechanism in alpaca.
... Function loss of MSTN is known to cause an increased muscle mass phenotype in several mammals, including mice, dogs, cattle and humans [13][14][15][16] . Fibroblast growth factor 5 (FGF5), a secreted signaling protein during the hair growth cycle, inhibits hair growth by blocking dermal papilla cell activation 17 , and is regarded as the causative gene underlying the angora phenotype (long hair coat) in mice 18 . Mutations in FGF5 underlie trichomegaly (excessively long eyelashes) in humans 19 , as well as being associated with hair length in other mammal species such as cats 20 , dogs 21 and donkeys 22 . ...
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Recent advances in the study of the CRISPR/Cas9 system have provided a precise and versatile approach for genome editing in various species. However, the applicability and efficiency of this method in large animal models, such as the goat, have not been extensively studied. Here, by co-injection of one-cell stage embryos with Cas9 mRNA and sgRNAs targeting two functional genes (MSTN and FGF5), we successfully produced gene-modified goats with either one or both genes disrupted. The targeting efficiency of MSTN and FGF5 in cultured primary fibroblasts was as high as 60%, while the efficiency of disrupting MSTN and FGF5 in 98 tested animals was 15% and 21% respectively, and 10% for double gene modifications. The on- and off-target mutations of the target genes in fibroblasts, as well as in somatic tissues and testis of founder and dead animals, were carefully analyzed. The results showed that simultaneous editing of several sites was achieved in large animals, demonstrating that the CRISPR/Cas9 system has the potential to become a robust and efficient gene engineering tool in farm animals, and therefore will be critically important and applicable for breeding.
... Both the FGF5 and FGF5S proteins function primarily through binding to FGF receptors 1 and 2. FGF5 actively inhibits cell proliferation and the synthesis of hair fibers during the anagen stage of the hair follicle growth cycle. FGF5S competitively binds the FGF receptor, antagonizing the inhibitory effects of FGF5 Suzuki et al., 2000;Ota et al., 2002). Studies in cats and dogs have indicated that FGF5 mutants were associated with altered hair length (Housley and Venta, 2006;Kehler et al., 2007;Cadieu et al., 2009;Dierks et al., 2013). ...
Article
The fibroblast growth factor 5 gene (FGF5) is a member of the FGF gene family, and represents a candidate gene for hair length because of its role in the regulation of the hair follicle growth cycle. In our current study, we cloned, sequenced, and characterized the full-length FGF5 cDNA of Chinese Merino sheep. We obtained the complete genomic sequence of the FGF5 gene from sheep blood samples, and compared it to other FGF5 sequences in GenBank. We found that the FGF5 gene spanned 21,743bp of genomic DNA, and consisted of 3 exons and 2 introns, both of which differed from those of a previously annotated FGF5 genomic sequence from sheep. We also identified a previously undescribed FGF5 mRNA splicing variant, FGF5S, and the western blot analysis showed that the molecular weights of the FGF5 (34kDa) and FGF5s (17kDa) proteins were consistent with the estimates based on the genomic and cDNA sequence data. We examined the expression of both FGF5 mRNAs in various tissues of sheep, and found that the expression of the FGF5S mRNA was restricted to the brain, spleen, and skin tissue. The single-nucleotide polymorphism analysis of the genomic sequence revealed 72 genetic variants of the FGF5 gene. Our findings provide insight into the functions of the FGF5 gene in Chinese Merino. Copyright © 2014 Elsevier B.V. All rights reserved.
... Localization of FGF5 in the Hair Follicle. FGF5 acts as a stimulus for catagen entry by binding to FGFR1 located within the dermal papilla, a mesenchymal signaling center located at the base of the hair follicle (17). However, the origin of FGF5 has not been clearly established within mouse skin. ...
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Mechanisms that regulate the growth of eyelashes have remained obscure. We ascertained two families from Pakistan who presented with familial trichomegaly, or extreme eyelash growth. Using a combination of whole exome sequencing and homozygosity mapping, we identified distinct pathogenic mutations within fibroblast growth factor 5 (FGF5) that underlie the disorder. Subsequent sequencing of this gene in several additional trichomegaly families identified an additional mutation in FGF5. We further demonstrated that hair fibers from forearms of these patients were significantly longer than hairs from control individuals, with an increased proportion in the growth phase, anagen. Using hair follicle organ cultures, we show that FGF5 induces regression of the human hair follicle. We have identified FGF5 as a crucial regulator of hair growth in humans for the first time, to our knowledge, and uncovered a therapeutic target to selectively regulate eyelash growth.
... Cultured DP cells stimulated with Fgf-2 produce factors that promote keratinocyte proliferation, and Fgf-5 can inhibit the production of these factors. While this shows that DP cells can respond to Fgf-5, further study is required to determine whether Fgf-5 acts directly on DP to terminate anagen (Ota et al. 2002). In a similar fashion, other pathways that can promote catagen, including transforming growth factor-b and neurotrophins, act in both epithelium and DP, and a specific role in DP has not been shown (Botchkarev and Paus 2003). ...
Article
The dermal papilla (DP) of the hair follicle is both a chemical and physical niche for epithelial progenitor cells that regenerate the cycling portion of the hair follicle and generate the hair shaft. Here, we review experiments that revealed the importance of the DP in regulating the characteristics of the hair shaft and frequency of hair follicle regeneration. More recent work showed that the size of this niche is dynamic and actively regulated and reduction in DP cell number per follicle is sufficient to cause hair thinning and loss. The formation of the DP during follicle neogenesis provides a context to contemplate the mechanisms that maintain DP size and the potential to exploit these processes for hair preservation or restoration.
... FGF-2 also has been reported to function in hair follicles. The FGF-1 and FGF-2 receptors (FGFR-1 and FGFR-2) were detected in DP cells, and FGF-2 enhances DPC-mediated outer root sheath cell proliferation (21). FGF-2 is essential for long term culture of dermal papilla cells and for the sphere formation that is a partial model of an intact DP, resulting in hair follicle induction (22,23). ...
Article
Hair regression and balding are distressing concerns for an increasing number of people due to changes in lifestyle and serious nutritional imbalances. Therapies for treatment of hair loss are needed. Among potential therapeutics, adenosine has been suggested as a potent regulator of hair growth. In this study, we investigated the effects of adenosine on hair follicles and dermal papilla (DP) cells, and the mechanism underlying the action of adenosine. Hair follicles are organs, including DP cells, that are responsible for the production of hair fibers by inducing and maintaining the hair growth phase (anagen). In a culture of DP cells in vitro, adenosine stimulated proliferation of DP cells by increasing thymidine uptake. Subsequently, adenosine activated and elongated the anagen phase by increasing the uptake of radiolabeled cysteine in an organ culture of mouse vibrissae hair follicles. We also confirmed that adenosine promoted the expression of several growth factors that are responsible for hair growth, including fibroblast growth factors (FGF)-7, FGF-2, insulin-like growth factor (IGF)-1, and vascular endothelial growth factor (VEGF) in a cDNA microarray with semi-quantitative RT-PCR. Transcriptional activation of β-catenin in DP cells was increased by adenosine in a luciferase assay. β-catenin is a co-activator of Wnt/β-catenin signaling that induces morphogenesis and differentiation of hair follicles and also acts to transactivate downstream signaling pathways, including the ERK pathway. Using Western blotting, we found that adenosine stimulated phosphorylation of ERK, CREB and AKT. These results suggest that adenosine stimulates growth of hair follicles by triggering the expression of growth factors and β-catenin, and by inducing their downstream target signaling pathways.
... c) Telogen: During this phase, both follicles and skin are at rest. This cycle is regulated by a variety of mediators, including several members of the insulin-like growth factor family.[1–3] ...
Article
Hair loss is seen as an irreversible process. Most research concentrates on how to elongate the anagen, reduce the negative factors of obstructing hair growth and improve the hair number and size. In our experiment, we tried to prove that the cow placenta extract can promote hair growth by elongating hair shaft and increasing hair follicle number. Cow placenta extract (CPE), water and minoxidil applied separately on the back of depilated B57CL/6 mice for the case, negative and positive control respectively. We checked the proliferation of cells which are resident in hair sheath, and the expression of a few growth factors which stimulate hair growth. Result shows that placenta extract more efficiently accelerates cell division and growth factor expression, by raising the insulin-like growth factor (IGF-1) mRNA and protein level to increase HF size and hair length. The extract is not a purified product; so, it is less effective than minoxidil, which is approved by the US FDA for the treatment of male pattern baldness. If refinement is done, the placenta extract would be a good candidate medicine for hair loss.
... In cell culture 2 FGF2 was signifi cantly down-regulated, indicating the anagen phase. Anyhow the signifi cant regulation of FGF2 was a surprise because several studies identifi ed this factor in the hair follicle but not in the hair papilla (Mitusi et al. 1997;Ota et al. 2002;Schlake et al. 2005). ...
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Doping with anabolic agents is a topic in sports where strength is crucial, e.g. sprinting, weight lifting and many more. Testosterone and its functional analogs are the drugs of choice taken as pills, creams, tape or injections to increase muscle mass and body performance, and to reduce body fat. Stanozolol (17β-hydroxy-17α-methyl-5α-androst-2-eno[3,2c]pyrazol) is a testosterone analogue with the same anabolic effect like testosterone but its ring structure makes it possible to take it orally. Therefore, stanozolol is one of the most frequently used anabolic steroids. Common verification methods for anabolic drugs exist, identifying the chemicals in tissues, like hair or blood samples. The idea of this feasibility study was to search for specific gene expression regulations induced by stanozolol to identify the possible influence of the synthetically hormone on different metabolic pathways. Finding biomarkers for anabolic drugs could be supportive of the existing methods and an additional proof for illegal drug abuse. In two separate cell cultures, human HFDPC (hair follicle dermal papilla cells) from a female and a male donor were treated with stanozolol. In the female cell culture treatment concentrations of 0 nM (control), 1 nM, 10 nM and 100 nM were chosen. Cells were taken 0 h, 6 h, 24 h and 48 h after stimulation and totalRNA was extracted. Learning from the results of the pilot experiment, the male cell culture was treated in 10 nM and 100 nM concentrations and taken after 0 h, 6 h, 24 h and 72 h. Using quantitative real-time RT-PCR expression of characteristics of different target genes were analysed. Totally 13 genes were selected according to their functionality by screening the actual literature and composed to functional groups: factors of apoptosis regulation were Fas Ligand (FasL), its receptor (FasR), Caspase 8 and Bcl-2. Androgen receptor (AR) and both estrogen receptors (ERα, ERβ) were summarized in the steroid receptor group. The growth factor group included the insulin like growth factor receptor (IGF1R) and growth hormone receptor (GHR). Fibroblast growth factor 2 (FGF2) and keratinocyte growth factor (FGF7) were summarized in the hair cycle factor group. 5α-Steroidreductases (SRD5A1, SRD5A2) represented the enzyme group. Three reference genes were taken for relative quantification: ubiquitin (UBQ), glycerinaldehyde-3-phsophate-dehydrogenase (GAPDH), and β-actin (ACTB). In cell culture 1 AR, FasR, FGF2 showed significant regulations within one treatment time, significant gene expressions over time were analysed for Caspase 8. In cell culture 2 AR, FasR and SRD5A2 were significantly regulated within one treatment time. In this feasibility study first biomarker for a screening pattern of anabolic agents could be identified providing the rationality to investigate modified, metabolic pathways in the whole hair follicle.
... By alternative splicing, FGF5 gene produces two transcripts, the full length FGF5 with complete three exons and short alternative transcript (FGF5s) without exon2 and part of exon3. The full length transcript which functions to inhibit the activation of derma papilla cells proliferation and synthesis of hair fiber during anagen, and promotes the transition from anagen to catagen (Ota et al., 2002). The FGF5s has no inhibitory function in hair growth, whereas it binds to the FGF receptor1 (FGFR1) and 2 (FGFR2) to compete the FGF5 binding and serves as an antagonist to inhibit FGF5 function (Ozawa et al., 1998). ...
Article
Fibroblast growth factor 5 (FGF5) has been recognized as an inhibitor to cease animal hair growth, while in contrary, FGF5 short alternative transcript (FGF5s) can induce hair growth by antagonizing FGF5 function. To investigate the role of FGF5s in wool growth in Chinese Merino sheep, we generated transgenic sheep of ectopic expression of FGF5s by injection of recombinant lentivirus into zygote. Totally 20 transgenic sheep were obtained and 12 were alive after birth. Characterization of the transgene revealed that the transgenic sheep showed variety of integrant, ranged from 2 to 11 copies of transgene. The ectopic expression of FGF5s was observed in all transgenic sheep. Further study on the effect of ectopic expression of FGF5s revealed that the wool length of transgenic sheep were significantly longer than that of non-transgenic control, with 9.17 cm of transgenic lambs versus 7.58 cm of control animals. Notably, besides the increase of wool length, the yearling greasy fleece weight was also concordantly greater than that of wild-type (p < 0.01), with 3.22 kg of transgenic sheep versus 2.17 kg of control lambs (p < 0.01) in average. Our results suggested that overexpression of FGF5s could stimulate wool growth and resulted in increase of wool length and greasy wool weight.
... In mouse, FGF5 mRNA is highly expressed in the hair follicle as two isoforms, identified as FGF5 and FGF5S (Suzuki et al., 2000), and the latter is due to the alternative splicing of exon 2 (Hattori et al., 1996). Both isoforms, through binding to FGF receptor 1 and 2, regulate the hair follicle growth cycle during the anagen stage: FGF5 actively inhibits cell proliferation and the synthesis of hair fibers, while FGF5S antagonizes the inhibitory effects of FGF5 through competi-tively binding the FGF receptors (Suzuki et al., 2000;Ota et al., 2002;He et al., 2016). ...
Article
In the Peruvian highlands livestock production is especially developed in the South and Center of the country between altitudes ranging from 2,200 to 4,500 meters above sea level. Community-owned pasture is the main source of animal feed in this region. Llama rearing is commonly combined with other species, such as alpacas and sheep. One way to technically evaluate livestock systems is by calculating technical parameters. These indicators allow to synthesize the information contained in the production records and to standardize the evaluation criteria of the production unit. The following technical parameters were used in this study: Annual average capital (AAC), gross birth rate (%GBR), real birth rate (%RBR), mortality (%M), harvest (%H), gross increase (%GI), real increase (% RI) and livestock efficiency (%LE). In the present study, the technical evaluation of livestock was carried out using the monthly records of llamas, alpacas and sheep of the Communal Cooperative San Pedro de Racco, located in Pasco Region, Peru. The evaluation period for llamas and alpacas was from 2012 to 2015, whereas for sheep the evaluation period was from January 2014 to December 2015. It was concluded that the Community Cooperative San Pedro de Racco presented values of technical parameters corresponding to a good technological level company, possibly due to the capacity development of human resources in terms of animal and grassland management.
... FGF-5 in hair follicle begins to be expressed at the end of anagen stage, it has a critical role in the transition of the hair follicle from anagen to catagen. FGF-5 promotes the cessation of anagen by binding to FGFR1 on the dermal papilla (Ota et al. 2002). Therefore, it was postulated that inhibition of FGF-5 can provide the therapeutic benefit for the short anagen syndrome (Higgins et al. 2014). ...
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Context: Eclipta prostrata L. (Asteraceae) (EP) has been widely used for the treatment of skin disease in Asian traditional medicine. Objective: This study investigates the potency of EP in promoting hair growth in vivo and in vitro. Materials and methods: C57BL/6N mice were divided into four groups (n = 4) as follows: control (topical treatment of normal saline), topical 3% minoxidil to the dorsal skin of mice for 14 days, and low (1 mg/day) and high (10 mg/day) doses of EP orally administered once a day for 14 days. Dorsal hairs of C57BL/6N mice were depilated to synchronize anagen induction. Hair growth activity was evaluated by gross and microscopic observations. Sections of dorsal skin were stained with haematoxylin and eosin. We also treated the various concentrations of EP (5, 10 and 50 μg/mL) for 24 h on the human dermal papilla cells (HDPs) and examined the effects of EP on the expression of FGF-7 and mTOR signalling. Results: EP enhanced the induction of anagen in the dorsal skin of mice, characterized by the appearance of inner root sheath along with hair shaft, the emergence of hair shaft through the epidermis. EP increased the expression of FGF-7, while decreased the level of FGF-5 in C57/BL6 mice. EP also increased the expression of FGF-7, activated the mTOR signalling in HDPs. Discussion and conclusions: These results suggest that EP has a potency to enhance the growth of hair follicle, promoting hair growth through regulation of FGF-7 and FGF-5.
... Then, several studies on the FGF5 gene demonstrated the relation to coat hair length in cats (Drögemüller et al., 2007;Shaffer et al., 2021), dogs (Dierks et al., 2013), humans (Higgins et al., 2014), donkeys (Legrand et al., 2014), hamsters (Yoshizawa et al., 2015), llamas (Daverio et al., 2017), guinea pigs (Yu et al., 2018), alpacas (Pallotti et al., 2018a,b), sheep (Zhang et al., 2019), and dromedary (Maraqa et al., 2021). It is currently regarded that FGF5 is overexpressed during the late anagen phase and serves as a crucial regulatory factor that promotes the anagento-catagen transition in the hair follicle cycle (Suzuki et al., 1998;Ota et al., 2002;He et al., 2016). The same selection target has been described in several cashmere goat investigations (Guo et al., 2019;Li et al., 2017). ...
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The FGF5 gene has been associated with the regulation of fibre length in mammals, including cashmere goats. A deletion variant at ~14 kb downstream of the FGF5 gene showed significant divergence between cashmere and non‐cashmere goats in previous studies. In this study, we designed specific primers to genotype the deletion variant. The results of gel electrophoresis and Sanger sequencing revealed that a 507‐bp deletion mutation is located at 95 454 685–95 455 191 of chromosome 6 in goats. Genotyping data from a large panel of 288 goats showed that the deletion at the FGF5 gene locus appeared to be associated with cashmere length. The deletion variant was close to fixation (frequency 0.97) in cashmere goats. Furthermore, electrophoretic mobility shift assays for evaluating DNA–protein interaction and mRNA expression levels of FGF5 suggested that the deletion variant may serve as a cis‐acting element by specifically binding transcription factors to mediate quantitative changes in FGF5 mRNA expression. Our study illustrates how a structural mutation of the FGF5 gene has contributed to the cashmere growth phenotype in domestic goats. The deletion mutation within the FGF5 gene could potentially serve as a molecular marker of cashmere growth in cashmere goat breeding.
... In mouse, FGF5 mRNA is highly expressed in the hair follicle as two isoforms, identified as FGF5 and FGF5S (Suzuki et al., 2000), and the latter is due to the alternative splicing of exon 2 (Hattori et al., 1996). Both isoforms, through binding to FGF receptor 1 and 2, regulate the hair follicle growth cycle during the anagen stage: FGF5 actively inhibits cell proliferation and the synthesis of hair fibers, while FGF5S antagonizes the inhibitory effects of FGF5 through competi-tively binding the FGF receptors (Suzuki et al., 2000;Ota et al., 2002;He et al., 2016). ...
... In mouse, FGF5 mRNA is highly expressed in the hair follicle as two isoforms, identified as FGF5 and FGF5S (Suzuki et al., 2000), and the latter is due to the alternative splicing of exon 2 (Hattori et al., 1996). Both isoforms, through binding to FGF receptor 1 and 2, regulate the hair follicle growth cycle during the anagen stage: FGF5 actively inhibits cell proliferation and the synthesis of hair fibers, while FGF5S antagonizes the inhibitory effects of FGF5 through competi-tively binding the FGF receptors (Suzuki et al., 2000;Ota et al., 2002;He et al., 2016). ...
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One experiment was conducted in order to test the utility of near-infrared spectroscopy (NIRS) to predict mohair quality, i.e.i) clean mohair content and ii) mean fibre diameter (MFD) in Angora goat fleece. A total of 397 mohair fleece samples were collected in 2016 within the framework of the French selection scheme including performance recording, fleece assessment and fibre measurements. Fleece samples were laboratory measured for clean mohair content and MFD by OFDA methodology, then scanned in a Petri dish using a NIRS portable instrument (LabSpec ® 5000; ASD Inc, Boulder, USA) by reflectance in the VIS and NIR regions (350to 2,500nm). Partial least square (PLS) regression was used to develop a number of calibration models between the spectral and reference measurements. Different mathematical treatments were used during model development. The methods studied were partial least squares regression (PLS) and first-derivative pretreatment + PLS.Cross validation was used to assess the performance and avoid overfitting of the models. NIR prediction of MFD gave a low R² (<0.70) whatever the calibration models. By using the firstderivative of the raw spectra in the NIR region (800 to 2,500nm), the calibration models gave a coefficient of determination in calibration (R²) of 0.81 for clean mohair content with a low relative error (3.5%). It is concluded that NIR reflectance spectroscopy can be used to predict clean mohair content with a good precision but not for determining mean fibre diameter. However the use of OFDA 2000 MFD measurement along the greasy staple can be used as an alternative. Thus NIR spectroscopy to predict clean mohair content and OFDA2000 to measure fibre diameter along greasy staple would be widely used to assist the French Angora goat breeding program and allowing a large reduced cost of fibre measurements.
... Zira genom düzenleme denildiğinde özgün bir bölgedeki baz çiftlerinin çıkarılması, değiştirilmesi veya eklenmesi akla gelmektedir. (Ota, 2002;Wang, 2015a). ...
Article
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Dünya nüfusunun hızla artmasıyla ortaya çıkan gıda ihtiyacını karşılayamayan tarımsal ürünlerin stok yetersizliği, iklim değişiklikleri sebebiyle çiftlik hayvanlarında ortaya çıkan adaptasyon güçlükleri ve yaygınlaşan çeşitli hastalıklar gibi problemler için her geçen gün yeni çözümler üretilmektedir. Son zamanlarda geliştirilen genom düzenleme tekniklerinin kullanımı ile bu sorunlara çözüm bulunabileceği bilim insanları tarafından kabul edilmektedir. Genom düzenleme, nükleazlar sayesinde genomda belirlenmiş bir yerden bölgeye özel DNA çift zincir kırıkları (DSB) oluşturduktan sonra homolog rekombinasyon (HDR) veya homolog olmayan rekombinasyon (NHEJ) yöntemlerinden biriyle çift zincir kırıkları tamir edilerek genom değişimini meydana getirebilme metodudur. Bu metotlar ile embriyo transfer teknolojisi birleştirilerek hayvan ıslahına uygulanmasında temel amaç, verim ve kaliteyi artırmanın yanında hayvan refahının arttırılması ve hastalıklara karşı direnç sağlanmasıdır. Bu çalışmada, genom düzenleme yöntemlerinin ve hayvancılıkta uygulama alanlarının açıklanması amaçlanmıştır.
... Interestingly, functional enrichment analyses indicated that the DEGs are over-represented in GO terms associated with muscle fiber development in MSTN-disrupted goats, and GO terms related to skin development and immune responses in FGF5-edited animals. MSTN is primarily thought to inhibit muscle differentiation and growth (Poncelet, 1997;Mosher et al., 2007), while FGF5 represses hair growth by blocking dermal papilla cell activation (Hebert et al., 1994;Ota et al., 2002). Therefore, it is plausible that disruption of these key regulators triggers multiple functional regulatory genes at posttranscriptional levels and eventually resulting in observed phenotypic changes. ...
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Unintended off-target mutations induced by CRISPR/Cas9 nucleases may result in unwanted consequences, which will impede the efficient applicability of this technology for genetic improvement. We have recently edited the goat genome through CRISPR/Cas9 by targeting MSTN and FGF5, which increased muscle fiber diameter and hair fiber length, respectively. Using family trio-based sequencing that allow better discrimination of variant origins, we herein generated offspring from edited goats, and sequenced the members of four family trios (gene-edited goats and their offspring) to an average of ∼36.8× coverage. This data was to systematically examined for mutation profiles using a stringent pipeline that comprehensively analyzed the sequence data for de novo single nucleotide variants, indels, and structural variants from the genome. Our results revealed that the incidence of de novo mutations in the offspring was equivalent to normal populations. We further conducted RNA sequencing using muscle and skin tissues from the offspring and control animals, the differentially expressed genes (DEGs) were related to muscle fiber development in muscles, skin development, and immune responses in skin tissues. Furthermore, in contrast to recently reports of Cas9 triggered p53 expression alterations in cultured cells, we provide primary evidence to show that Cas9-mediated genetic modification does not induce apparent p53 expression changes in animal tissues. This work provides adequate molecular evidence to support the reliability of conducting Cas9-mediated genome editing in large animal models for biomedicine and agriculture.
... Many isoforms of KRTAP were upregulated in white compared with black hairs 42,43 . The expression level of fibroblast growth factor 5 (FGF5), an upstream inhibitor of KRT and KRTAP synthesis, was reduced in white compared with black hairs 44,45 . In contrast, the expression level of FGF7, an upstream stimulator of KRT and KRPAP synthesis, was increased in white compared with black hairs 46,47 . ...
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Hair graying is an obvious sign of human aging. Although graying has been investigated extensively, the mechanism remains unclear. Here, we reviewed previous studies on the mechanism of graying and seek to offer some new insights. The traditional view is that hair graying is caused by exhaustion of the pigmentary potential of the melanocytes of hair bulbs. Melanocyte dysfunction may be attributable to the effects of toxic reactive oxygen species on melanocyte nuclei and mitochondria. A recent study suggests that bulge melanocyte stem cells (MSCs) are the key cells in play. Graying may be caused by defective MSC self-maintenance, not by any deficiency in bulbar melanocytes. Our previous study suggested that graying may be principally attributable to active hair growth. Active hair growth may produce oxidative or genotoxic stress in hair bulge. These internal stress may cause eventually depletion of MSC in the hair follicles. Taken together, hair graying may be caused by MSC depletion by genotoxic stress in the hair bulge. Hair graying may also be sometimes caused by dysfunction of the melanocytes by oxidative stress in the hair bulb. In addition, hair graying may be attributable to MSC depletion by active hair growth. Copyright © The Korean Dermatological Association and The Korean Society for Investigative Dermatology..
... Various growth factors and cytokines are secreted from DPCs, which act as potent mitogenic factors and play important roles in follicular development and vasculogenesis. The resultant effects of growth factors and cytokines on proliferation of cultured DPCs and other mechanistic arms have been studied by researchers [109][110][111][112][113][114][115][116][117][118][119][120][121][122]. ...
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Hair disorders such as hair loss (alopecia) and androgen dependent, excessive hair growth (hirsutism, hypertrichosis) may impact the social and psychological well‐being of an individual. Recent advances in understanding the biology of hair have accelerated the research and development of novel therapeutic and cosmetic hair growth agents. Preclinical models aid in dermocosmetic efficacy testing and claim substantiation of hair growth modulators. The in vitro models to investigate hair growth utilize the hair follicle Dermal Papilla cells (DPCs), specialized mesenchymal cells located at the base of hair follicle that play essential roles in hair follicular morphogenesis and postnatal hair growth cycles. In this review, we have compiled and discussed the extensively reported literature citing DPCs as in vitro model to study hair growth promoting and inhibitory effects. A variety of agents such as herbal and natural extracts, growth factors and cytokines, platelet‐rich plasma, placental extract, stem cells and conditioned medium, peptides, hormones, lipid‐nanocarrier, light, electrical and electromagnetic field stimulation, androgens and their analogs, stress‐serum and chemotherapeutic agents etc. have been examined for their hair growth modulating effects in DPCs. Effects on DPCs’ activity were determined from untreated (basal) or stress induced levels. Cell proliferation, apoptosis and secretion of growth factors were included as primary end‐point markers. Effects on a wide range of biomolecules and mechanistic pathways that play key role in the biology of hair growth were also investigated. This consolidated and comprehensive review summarizes the up‐to‐date information and understanding regarding DPCs based screening models for hair growth and may be helpful for researchers to select the appropriate assay system and biomarkers. This review highlights the pivotal role of DPCs in the forefront of hair research as screening platforms by providing insights into mechanistic action at cellular level, which may further direct the development of novel hair growth modulators. This article is protected by copyright. All rights reserved.
Article
Several growth factors and cytokines have been shown to be involved in normal hair cycling as well as in androgenetic alopecia (AGA). However, the molecular cascades in AGA downstream from androgen receptor activation are far from being fully elucidated. We sought to determine the difference in the protein expression of growth factors/cytokines in balding vs. nonbalding scalp specimens from the same individuals affected with AGA. Balding and nonbalding scalp specimens were collected from four men with pattern baldness. Dermal papilla (DP) cells were isolated and cultured. Quantifying the protein expression of growth factors and cytokines expressed by these cells was performed using Quantibody® Human Growth Factor Array-1 (RayBiotech, Inc., Norcross, GA, U.S.A.). Brain-derived nerve factor (BDNF) protein expression was upregulated by approximately 12-fold in supernatants obtained from balding as compared with nonbalding DP cells (P < 0·001). Expression of neurotrophin-3 and of β-nerve growth factor was also upregulated. On the other hand, protein expression of insulin-like growth factor-1 and its binding proteins as well as of the vascular endothelial growth factor family were significantly downregulated in the balding scalp. Neurotrophic factors, especially BDNF, may be important in mediating the effects of androgens on hair follicles, serving as a negative regulatory control signal. Further studies may lead to novel pharmacological interventions in AGA.
Article
Hair greying is an obvious sign of ageing in humans. White (nonpigmented) hair is thicker than black (pigmented) hair. The growth rate of white hair is also significantly higher than that of black hair. However, the mechanism underlying this is largely unknown. To examine the association between hair greying and hair growth patterns by evaluating expression of the genes or proteins related to hair growth in white and black hairs. Morphological characteristics were observed in eyebrow and scalp hairs. The differential expression of genes was analysed in black and white hairs from human scalp by a microarray analysis. Reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry for genes and proteins related to hair growth were performed in black and white hairs. Keratin and keratin-associated protein (KRTAP) genes in white hair were upregulated at least two-fold in comparison with black hair in a microarray analysis. Upregulation of selected keratin genes and KRTAP4 isoform genes in white hair was validated by RT-PCR. Immunoreactivity for KRT6, KRT14/16 and KRT25 was increased in the hair follicle of white hair compared with black hair. Gene expression of fibroblast growth factor 5 (FGF5) was downregulated in white hair compared with black hair. However, gene expression of FGF7 was upregulated in white hair compared with black hair. Expression of genes and proteins associated with active hair growth is upregulated in white (nonpigmented) hair compared with black (pigmented) hair. These results suggest that hair greying is associated with active hair growth.
Article
The article presents current data on the role growth factors play in hair physiology. Based on a review of literature, the authors described the role growth factors play for initiating, suppressing the growth and differentiating hair follicles. According to them, each morphologic development stage of hair follicles is characterized by its own factor expression pattern. Referring to experimental and clinical studies, the authors describe the role some growth factors play for mechanisms promoting the development of androgynous and focal alopecia.
Chapter
Androgenetic Alopecia (AGA) is by far the most common cause of hair loss in men, and its high prevalence has been reported in detail for many decades [1]. Several different terms in international medical bibliography have been suggested by several authors, such as androgenic alopecia, male pattern baldness, androgen-dependent alopecia, common baldness, and genetic hair loss. However, the term “Androgenetic Alopecia” is considered the most appropriate since it summarizes the etiology of the condition, with the term “andro-” referring to the hormonal and “-genetic”, implying the inherited parameter of its pathogenesis.
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The hair follicle (HF) is a multi-tissue mini-organ that self-renews periodically. However, the cellular organisation of this much-studied model is not fully understood. The structures of the outer layer and of the bulb, which ensures HF growth, have not been completely established. To clarify these points, we have conducted in vivo clonal analyses with 3D imaging in mice. The upper two-thirds of the HF outer layer consists of two clonally unrelated groups of cells that exhibit different modes of growth. They correspond to the basal outer root sheath (ORS) and the companion layer (Cp). The basal ORS has an unusual anisotropic mode of growth from a suprabulbar zone, which we named the privileged proliferation zone. The Cp has a stem/transient-amplifying mode of growth and is shown to be an HF internal structure. Furthermore, we describe an additional element, the bulb outer layer, which is contiguous and shares markers (e.g. Lgr5) with the basal ORS but is formed by a separate lineage that belongs neither to the ORS nor Cp lineage. It represents a novel element with proximal cells that are contiguous with the germinative layer in the bulb. In reference to its shape and position we named it the lower proximal cup (LPC). These clonal hierarchies reveal a novel model of HF organisation and growth based on two major entities: the basal ORS and the LPC plus the seven internal layers.
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Wool is the critical textile raw material which is produced by the hair follicle of sheep. Therefore, it has important implications to investigate the molecular mechanism governing hair follicle development. Due to high cellular heterogeneity as well as the insufficient cellular, molecular, and spatial characterization of hair follicles on sheep, the molecular mechanisms involved in hair follicle development and wool curvature of sheep remains largely unknown. Single-cell RNA sequencing (scRNA-seq) technologies have made it possible to comprehensively dissect the cellular composition of complex skin tissues and unveil the differentiation and spatial signatures of epidermal and hair follicle development. However, such studies are lacking so far in sheep. Here, single-cell suspensions from the curly wool and straight wool lambskins were prepared for unbiased scRNA-seq. Based on UAMP dimension reduction analysis, we identified 19 distinct cell populations from 15,830 single-cell transcriptomes and characterized their cellular identity according to specific gene expression profiles. Furthermore, novel marker gene was applied in identifying dermal papilla cells isolated in vitro . By using pseudotime ordering analysis, we constructed the matrix cell lineage differentiation trajectory and revealed the dynamic gene expression profiles of matrix progenitors' commitment to the hair shaft and inner root sheath (IRS) cells. Meanwhile, intercellular communication between mesenchymal and epithelial cells was inferred based on CellChat and the prior knowledge of ligand–receptor pairs. As a result, strong intercellular communication and associated signaling pathways were revealed. Besides, to clarify the molecular mechanism of wool curvature, differentially expressed genes in specific cells between straight wool and curly wool were identified and analyzed. Our findings here provided an unbiased and systematic view of the molecular anatomy of sheep hair follicle comprising 19 clusters; revealed the differentiation, spatial signatures, and intercellular communication underlying sheep hair follicle development; and at the same time revealed the potential molecular mechanism of wool curvature, which will give important new insights into the biology of the sheep hair follicle and has implications for sheep breeding.
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Objectives: The hair follicle is composed of more than 20 kinds of cells, and mesoderm derived dermal papilla cells and keratinocytes cooperatively contribute hair growth via Wnt/β-catenin signaling pathway. We are to investigate β-catenin expression and regulatory mechanism by CBD in alopecia hair tissues and dermal papilla cells. Methods: We performed structural and anatomical analyses on alopecia patients derived hair tissues using microscopes. Pharmacological effect of CBD was evaluated by β-catenin expression using RT-PCR and immunostaining experiment. Results: Morphological deformation and loss of cell numbers in hair shaft were observed in alopecia hair tissues. IHC experiment showed that loss of β-catenin expression was shown in inner shaft of the alopecia hair tissues, indicating that β-catenin expression is a key regulatory function during alopecia progression. Consistently, β-catenin expression was decreased in testosterone or PMA treated dermal papilla cells, suggesting that those treatments are referred as a model on molecular mechanism of alopecia using dermal papilla cells. RT-PCR and immunostaining experiments showed that β-catenin expression was decreased in RNA level, as well as decreased β-catenin protein might be resulted from ubiquitination. However, CBD treatment has no changes in gene expression including β-catenin, but the decreased β-catenin expression by testosterone or PMA was restored by CBD pretreatment, suggesting that potential regulatory effect on alopecia induction of testosterone and PMA. Conclusion: CBD might have a modulating function on alopecia caused by hormonal or excess of signaling pathway, and be a promising application for on alopecia treatment.
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Although hair forms (straight, curly, wavy, etc.) are present in apparently infinite variations, each fibre can be reduced to a finite sequence of tandem segments of just three types: straight, bent/curly, or twisted. Hair forms can thus be regarded as resulting from genetic pathways that induce, reverse or modulate these basic curvature modes. However, physical interconversions between twists and curls demonstrate that strict one-to-one correspondences between them and their genetic causes do not exist. Current hair-curvature theories do not distinguish between bending and twisting mechanisms. We here introduce a multiple papillary centres (MPC) model which is particularly suitable to explain twisting. The model combines previously known features of hair cross-sectional morphology with partially/completely separated dermal papillae within single follicles, and requires such papillae to induce differential growth rates of hair cortical material in their immediate neighbourhoods. The MPC model can further help to explain other, poorly understood, aspects of hair growth and morphology. Separate bending and twisting mechanisms would be preferentially affected at the major or minor ellipsoidal sides of fibres, respectively, and together they exhaust the possibilities for influencing hair-form phenotypes. As such they suggest dialectic for hair-curvature development. We define a natural-dialectic (ND) which could take advantage of speculative aspects of dialectic, but would verify its input data and results by experimental methods. We use this as a top-down approach to first define routes by which hair bending or twisting may be brought about and then review evidence in support of such routes. In particular we consider the wingless (Wnt) and mammalian target of rapamycin (mTOR) pathways as paradigm pathways for molecular hair bending and twisting mechanisms, respectively. In addition to the Wnt canonical pathway, the Wnt/Ca(2+) and planar cell polarity (PCP) pathways, and others, can explain many alternatives and specific variations of hair bending phenotypes. Mechanisms for hair papilla budding or its division by bisection or fission can explain MPC formation. Epithelial-to-mesenchymal (EMT) and mesenchymal-to-epithelial (MET) transitions, acting in collaboration with epithelial-mesenchymal communications are also considered as mechanisms affecting hair growth and its bending and twisting. These may be treated as sub-mechanisms of an overall development from neural-crest stem cell (NCSC) lineages to differentiated hair follicle (HF) cell types, thus providing a unified framework for hair growth and development.
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This study was conducted to evaluate the effects of Sargassum muticum extract and apo-9′-fucoxanthinone, a principal component of S. muticum, on hair growth. When rat vibrissa follicles were treated with S. muticum extract for 21 d, the hair-fiber lengths for the vibrissa follicles increased significantly. Treatment with the S. muticum extract and the EtOAc fraction of the S. muticum extract markedly increased the proliferation of dermal papilla cells (DPCs) and decreased the 5α-reductase activity. In addition, the EtOAc fraction of the S. muticum extract significantly promoted anagen initiation in C57BL/6 mice. Especially, apo-9′-fucoxanthinone, an active constituent from the S. muticum extract, caused an increase in DPC proliferation and a decrease in 5α-reductase activity. To elucidate the molecular mechanisms of apo-9′-fucoxanthinone on the proliferation of DPCs, we examined the level of various signaling proteins. Apo-9′-fucoxanthinone increased the level of vascular endothelial growth factor receptor-2 (VEGF-R2), Wnt/β-catenin signaling proteins such as phospho(ser9)-glycogen synthase kinase-3β (GSK-3β) and phospho(ser552)-β-catenin, whereas apo-9′-fucoxanthinone did not affect the transforming growth factor-β (TGF-β) signaling proteins such as Smad2/3. These results suggest that apo-9′-fucoxanthinone from S. muticum could have the potential for hair growth with DPC proliferation via the activation of Wnt/β-catenin signaling and the VEGF-R2 pathway.
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Despite their name, fibroblast growth factors (FGFs) are multifunctional regulators affecting a wide variety of physiological events. This review summarizes our recent studies on FGFs from mechanistic, physiological and application-oriented viewpoints. These include studies on the importance of βKlotho and glycosaminoglycans for the signaling of hormonal FGFs (FGF21 and FGF19); the physiological role of a paracrine FGF (FGF18) in hair cycle regulation; and the development of a stable, chimeric FGF protein composed of FGF1 and FGF2 domains suitable for radioprotection.
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As a subjective mood-related disorder with an unclear mechanism, depression has many problems in its diagnosis, which offers great space and value for research. At present, the methods commonly used to judge whether an animal model of depression has been established are mainly by biochemical index detection and behavioral tests, both of which inevitably cause stress in animals. Stress-induced hair growth inhibition has been widely reported in humans and animals. The simplicity of collecting hair samples and the observable state of hair growth has significant advantages; we tried to explore whether the parameters related to hair growth could be used as auxiliary indicators to evaluate a depression model in animals. The length and weight of the hair were calculated. Correlation analysis was conducted between the depressive behavioral results and the glucocorticoid levels in hair and serum. Learned helplessness combined with chronic restraint stress, and chronic unpredictable stress in the animal were detectable by superficial observation, weight ratio, and length of hair, and follicular development scores were significantly reduced compared to the control. The hair growth parameters of rats with depression, the rise in corticosterone, and the corresponding changes in behavioral parameters were significantly correlated. The neurotrophic factors, glucocorticoid-receptor (GR), brain-derived neurotrophic factor (BDNF), fibroblast growth factor 2 (FGF2), and fibroblast growth factor 5 (FGF5), are associated with depression and hair growth. Significant differences were detected between the stress and control groups, suggesting that the mechanism underlying the stress-phenomenon inhibition of hair growth may be related to growth factor mediation.
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The ability to alter single bases without DNA double strand breaks provides a potential solution for multiplex editing of livestock genomes for quantitative traits. Here, we report using a single base editing system, Base Editor 3 (BE3), to induce nonsense codons (C-to-T transitions) at four target sites in caprine FGF5. All five progenies produced from microinjected single-cell embryos had alleles with a targeted nonsense mutation and yielded expected phenotypes. The effectiveness of BE3 to make single base changes varied considerably based on sgRNA design. Also, the rate of mosaicism differed between animals, target sites, and tissue type. PCR amplicon and whole genome resequencing analyses for off-target changes caused by BE3 were low at a genome-wide scale. This study provides first evidence of base editing in livestock, thus presenting a potentially better method to introgress complex human disease alleles into large animal models and provide genetic improvement of complex health and production traits in a single generation.
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Rabbit fur characteristics are primarily genetically determined traits. We used Illumina high‐throughput sequencing technology to assess gene expression in the skin tissues of rabbits derived from a cross between Wanxi Angora rabbits and Rex rabbits, which exhibit differential characteristics of short and long wool respectively, to investigate molecular mechanisms related to wool length determination. To identify key regulatory genes involved in rabbit wool length, genes that were differentially expressed between the long‐ and short‐wool rabbits based on a P‐value < 0.05 and log2|fold change| > 1 were characterized. A total of 798 genes were up‐regulated and 523 were down‐regulated in the long‐wool group compared to expression levels in the short‐wool group, and these genes were annotated with GO terms and KEGG pathways, revealing wool‐development‐related biological functions. The Wnt, Hedgehog and TGF‐β signaling pathways, which are related to cell proliferation, fibroblast proliferation and hair follicle regulation respectively, were identified. The expression levels of eight genes were validated by RT‐qPCR. In addition, an interaction network was constructed to show the regulatory relationships among the differentially expressed genes. In this study, we found that FGF5, WNT5A, BMP4 and BMP7 showed significant differential expression between the two groups. These transcriptomic profiling results provide comprehensive gene expression information for improving understanding of the molecular mechanisms involved in the growth and development of rabbit wool.
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Fibroblast growth factor (FGF) 5 regulates the development and periodicity of hair follicles, which can affect hair traits. Loss-of-function mutations associated with long-hair phenotypes have been described in several mammalian species. Sheep is an important economic animal, however, the evolution characterizations and biological mechanism of oFGF5 (Ovis aries FGF5) gene are still poorly understood. In this study, oFGF5 gene was obtained by resequencing the whole genome of three Dorper sheep and RACE of two Kazakh sheep FGF5. We proposed FGF5 was phylogenetically related to FGF4 family and oFGF5 clearly orthologed to goat FGF5. Six loci were found from the positive selection results of FGF5 and half of them located on signal peptide. The basically similar rates of function-altering substitutions in sheep and goat lineage and the rest of the mammalian lineage of 365 SNPs indicated that the FGF5 gene was quite conservative during evolution. Homology modeling of the oFGF5 suggested that it has a highly conserved FGF superfamily domain containing 10 β-strands. Furthermore, the protein–protein docking analysis revealed that oFGF5 have the potential to form heterodimers with oFGFR1, the predicted interaction interface of FGF5-FGFR1 heterodimer was formed mainly by residues from FGF superfamily domain. Our observations suggested the evolutionary and structural biology features of oFGF5 might be relevant to its function about hair follicle development and modulating hair growth, and we confirmed our speculation by using the FGF5 gene editing sheep produced by CRISPR/Cas9 technology.
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Objectives: Recently, thread-embedding therapy (TET) has been widely applied in Korean medicine for cosmetic purposes such as reducing skin wrinkles. An inserted thread was reported to have induced continuous stimulation, followed by support for connective tissue regeneration. However, the potential role of TET in hairgrowth has not yet been reported. Methods: We designed this study to evaluate whether TET has a hair-growth-promoting effect. C57 black 6 (C57BL/6) mice were divided into three groups: normal saline-treated, minoxidil-treated, and thread-embedded groups. Normal saline or 5% minoxidil was topically sprayed on the dorsal skin of the mice once a day for 16 days. Medical threads were embedded into the dorsal skin of the mice in a single application. Hair growth activity was evaluated by using dermoscopic and microscopic observations. Sections of the dorsal skin were stained with hematoxylin and eosin. Expressions of bromodeoxyuridine (BrdU), proliferating cell nuclear antigen (PCNA), fibroblast growth factor-7 (FGF-7), and fibroblast growth factor-5 (FGF-5) were detected by using immunohistochemical staining. A reverse transcription-polymerase chain reaction (RT-PCR) analysis was adopted to measure the messenger RNA (mRNA) expressions of FGF-7 and FGF-5. Results: TET enhanced anagen development in the hair follicles of C57BL/6 mice. The expressions of BrdU and PCNA, both of which imply active cellular proliferation, were increased by using TET. Moreover, TET increased the expression of FGF-7, an anagen-inducing growth factor, while decreasing the expression of FGF-5, an anagen-cessation growth factor, both at the protein and the mRNA levels. Conclusion: TET enhanced hair re-growth in C57BL/6 mice. TET regulated the expressions of anagen-associated growth factors and activated the proliferation of hair follicular cells in depilated skin lesions. Considering its long-lasting effect, TET may be a good alternative therapeutic for the treatment of alopecia.
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Advances in skin regeneration have resulted in techniques and products that have allowed regeneration of both the dermis and epidermis. Yet complete skin regeneration requires the adnexal skin structures. Thus it is crucial to understand the regenerative potential of hair follicles where genetic, nutritional, and hormonal influences have important effects and are critical for skin regeneration. The follicular stem cell niche serves as an anatomical compartment, a structural unit, a functional integrator, and a dynamic regulator necessary to sustain internal homeostasis and respond to outside stimuli. In particular, mechanics such as pressure, compression, friction, traction, stretch, shear, and mechanical wounding can influence hair loss or growth. Relevant niche signaling pathways such as Wnt, bone morphogenetic protein, fibroblast growth factor, Shh, and Notch may yield potential targets for therapeutic interventions.
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La familia de retinoblastoma agrupa a tres miembros, pRb, p107 y p130, que han sido implicados en los procesos de proliferación, diferenciación y carcinogénesis. Debido a la ausencia de estudios en modelos animales in vivo acerca del papel de esta familia en la homeostasis de la piel, se analizó la epidermis de diferentes modelos animales deficientes en una o varias de estas proteínas.La deficiencia conjunta de p107 y p130 provoca alteraciones en la diferenciación terminal y retraso en el desarrollo de los folículos pilosos. Los defectos observados en los folículos pueden ser debidos a alteraciones en la vía de señalización del morfógeno BMP4. Por otro lado, se observó la existencia de una acumulación ectópica de b-catenina en el núcleo de los queratinocitos basales de la epidermis debida a la sobreexpresión de Frat. Los animales deficientes en pRb son letales embrionarios. Por ello, para analizar el papel de esta proteína en la epidermis adulta se usó la tecnología Cre/Loxp que permite la deleción de una región génica de forma específica de tejido. La pérdida de pRb y la progresiva pérdida de alelos de p107 producía un fenotipo de hiperplasia que ocurría de forma dosis-dependiente. Este fenotipo se relacionó con graves alteraciones en los procesos de proliferación y diferenciación. Además, se determinó que pRb tiene un papel esencial en el control del estado postmitótico del queratinocito diferenciado. Las alteraciones observadas en la diferenciación se deben a un control defectuoso de la actividad E2F.Por último se analizó el papel de pRb en la carcinogénesis de piel. En un protocolo de carcinogénesis química se observó que, en ausencia de pRb, se producían menos tumores aunque más malignos. Además, la tasa apoptótica de los papilomas obtenidos era mayor que la observada en sus respectivos tumores controles. Ello se debía a una estabilización selectiva de p53.
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The heparin-binding growth factors include a family of seven structurally related proteins that can potentially interact with four known high affinity receptors. We have cloned the murine homologues of fibroblast growth factor receptors 1 and 3 (mFR1 and mFR3). To define the ligand specificity of these receptors, we have characterized their binding properties with respect to acidic and basic fibroblast growth factors (aFGF and bFGF, respectively) and their biologic activity with respect to aFGF, bFGF, FGF-4/K-FGF, and FGF-5. Unlike mFR1, which binds both aFGF and bFGF, mFR3 preferentially binds aFGF. mFR3-mediated mitogenicity also favors aFGF and FGF-4 with a 10-12-fold lower response to bFGF and no response to FGF-5. Both receptor binding and growth factor-mediated mitogenicity are dependent on heparin. Heparin-binding growth factor activity can thus be regulated by proteoglycans and by the type of FGF receptor expressed on the target cell.
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An androgen-dependent mouse mammary carcinoma cell line (SC-3) requires androgen for growth stimulation. We have shown previously that androgen acts on SC-3 cells to induce secretion of a fibroblast growth factor (FGF)-like growth factor, which in turn stimulates growth of the cells in an autocrine manner. In this study, the androgen-induced growth factor (AIGF) was purified from a conditioned medium of SC-3 cells stimulated with testosterone. cDNA cloning of AIGF by use of its partial amino acid sequence data revealed that AIGF is a distinctive FGF-like growth factor. An AIGF cDNA (pSC17) encodes a 215-amino acid protein with a putative signal peptide, which shares 30-40% homology with known members of the FGF family. The AIGF mRNA was markedly induced by 10 nM testosterone in Northern blot analysis. Expression of AIGF cDNA in mammalian cells clearly showed remarkable stimulatory effects of AIGF on growth of SC-3 cells in the absence of androgen. Thus, it is clear that the androgen-induced growth of SC-3 cells is mediated in an autocrine manner by AIGF, which is secreted by the tumor cells themselves in response to hormonal stimuli.
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A growth factor specific for epithelial cells was identified in conditioned medium of a human embryonic lung fibroblast cell line. The factor, provisionally termed keratinocyte growth factor (KGF) because of its predominant activity on this cell type, was purified to homogeneity by a combination of ultrafiltration, heparin-Sepharose affinity chromatography, and hydrophobic chromatography on a C4 reversed-phase HPLC column. KGF was both acid and heat labile and consisted of a single polypeptide chain of approximately 28 kDa. Purified KGF was a potent mitogen for epithelial cells, capable of stimulating DNA synthesis in quiescent BALB/MK epidermal keratinocytes by greater than 500-fold with activity detectable at 0.1 nM and maximal at 1.0 nM. Lack of mitogenic activity on either fibroblasts or endothelial cells indicated that KGF possessed a target cell specificity distinct from any previously characterized growth factor. Microsequencing revealed an amino-terminal sequence containing no significant homology to any known protein. The release of this growth factor by human embryonic fibroblasts raises the possibility that KGF may play a role in mesenchymal stimulation of normal epithelial cell proliferation.
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A major proportion of carcinomas induced by mouse mammary tumour virus (MMTV) show evidence for proviral activation of a cellular gene, int-2, on chromosome 7. The sequence of 7869 bp of DNA spanning the transcription unit of int-2 was determined and compared with that of a series of int-2-specific cDNA clones derived from mammary tumour RNA. The predicted positions of intron-exon boundaries, established by alignment of cDNA and chromosomal DNA sequences, indicate that the gene comprises at least three exons. An open reading frame capable of encoding a protein of 245 amino acids with an estimated mol. wt of 27 kd, is flanked by substantial non-coding segments at both 5' and 3' ends. Comparison of the chromosomal DNA sequence and the predicted amino acid sequence with available data-bases has revealed no homology to other known genes. These results are discussed in relation to the status of int-2 as a candidate proto-oncogene.
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We recently reported that the protein encoded in a novel human oncogene isolated from Kaposi sarcoma DNA was a growth factor with significant homology to basic and acidic fibroblast growth factors (FGFs). To study the properties of this growth factor (referred to as K-FGF) and the mechanism by which the K-fgf oncogene transforms cells, we have studied the production and processing of K-FGF in COS-1 cells transfected with a plasmid encoding the K-fgf cDNA. The results show that, unlike basic and acidic FGFs, the K-FGF protein is cleaved after a signal peptide, glycosylated, and efficiently secreted as a mature protein of 176 or 175 amino acids. Inhibition of glycosylation impaired secretion, and the stability of the secreted K-FGF was greatly enhanced by the presence of heparin in the cultured medium. We have used the conditioned medium from transfected COS-1 cells to test K-FGF biological activity. Similar to basic FGF, the K-FGF protein was mitogenic for fibroblasts and endothelial cells and induced the growth of NIH 3T3 mouse cells in serum-free medium. Accordingly, K-fgf-transformed NIH 3T3 cells grew in serum-free medium, consistent with an autocrine mechanism of growth. We have also expressed the protein encoded in the K-fgf protooncogene in COS-1 cells, and it was indistinguishable in its molecular weight, glycosylation, secretion, and biological activity from K-FGF. Taken together, these results suggest that the mechanism of activation of this oncogene is due to overexpression rather than to mutations in the coding sequences.
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We previously described the isolation of a human oncogene which had acquired transforming potential by a DNA rearrangement accompanying transfection of NIH 3T3 cells with human tumor DNA (X. Zhan, A. Culpepper, M. Reddy, J. Loveless, and M. Goldfarb, Oncogene 1:369-376, 1987). We now term this oncogene the FGF-5 gene, since it specifies the fifth documented protein related to fibroblast growth factors (FGFs. Two regions of the FGF-5 sequence, containing 122 of its 267 amino acid residues, were 40 to 50% homologous to the sequences of acidic and basic FGFs as well as to the sequences of the FGF-related oncoproteins int-2 and hst/KS3. The FGF-5 gene bears the three exon structures typical for members of this family. FGF-5 was found to be expressed in the neonatal brain and in 3 of the 13 human tumor cell lines examined. Several experiments strongly suggested that FGF-5 is a growth factor with properties common to those of acidic and basic FGFs. The rearrangement which activated the FGF-5 gene during DNA transfection had juxtaposed a retrovirus transcriptional enhancer just upstream from the native promoter of the gene.
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Clones encoding the angiogenic endothelial cell mitogen, basic fibroblast growth factor (FGF), have been isolated from human cDNA libraries made from kidney, fetal heart, fetal liver, term placenta, and a breast carcinoma. Basic FGF cDNA clones are present in these libraries at very low levels when compared to the quantity of the growth factor in the tissues. This observation, combined with the fact that several of the clones represent unspliced transcripts, suggests that cytoplasmic basic FGF mRNA is unstable and that the protein is stored in tissues. The amino acid sequence of human basic FGF, deduced from the sequence of these cDNAs and from genomic clones, is 99% homologous to that of bovine basic FGF, implying a strong selection pressure for maintenance of function and structure. As with the bovine factor, human basic FGF does not appear to have a signal peptide sequence. Southern blot analysis of human genomic DNA and mapping of the cloned gene shows that there is only one basic FGF gene. All of the basic, heparin-binding endothelial cell mitogens of similar amino acid composition that have been described must therefore be products of this single gene.
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The growth and development of hair follicles is influenced by a number of different growth factors and cytokines, particularly members of the fibroblast growth factor (FGF) family. Keratinocyte growth factor (KGF or FGF-7) is a recently identified 28-kd member of the FGF family that induces proliferation of a wide variety of epithelial cells, including keratinocytes within the epidermis and dermal adnexa. Because KGF induces marked proliferation of keratinocytes, and both KGF and KGF receptor (KGFR) mRNA are expressed at high levels in skin, we sought to localize KGF and KGFR in skin by in situ hybridization. KGFR mRNA was relatively strongly expressed by keratinocytes in the basilar epidermis as well as throughout developing hair follicles of rat embryos and neonates. KGF mRNA was expressed at lower levels than was KGFR but could be localized to follicular dermal papillae in rat embryos and neonates. These results prompted us to investigate the effects of KGF on hair follicles in two distinct murine models of alopecia. In the first model, recombinant KGF (rKGF) induced dose-dependent hair growth over most of the body in nu/nu athymic nude mice when administered intraperitoneally or subcutaneously over 17 to 18 days. When administered subcutaneously, rKGF induced the most extensive hair growth at the sites of injection. Histologically, rKGF induced marked follicular and sebaceous gland hypertrophy, a normalization of the nu/nu follicular keratinization defect, and an increase in follicular keratinocyte proliferation as assessed by bromodeoxyuridine labeling. In the second model, a neonatal rat model of cytosine arabinoside chemotherapy-induced alopecia in which interleukin-1, epidermal growth factor, and acidic FGF have all demonstrated some degree of alopecia cytoprotection, rKGF induced a dose-dependent cytoprotective effect, abrogating as much as 50% of the alopecia in this model when administered beginning 1 day before the onset of chemotherapy. Taken together, these data suggest that KGF is an important endogenous mediator of normal hair follicle growth, development, and differentiation.
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Keratinocyte growth factor (KGF) is a member of the fibroblast growth factor (FGF) family. Synthesized by cells of the dermal component of skin, KGF's potent mitogenic activity is on the epidermal component, which harbors the receptors for this factor. To explore the possible role of KGF in mesenchymal-epithelial interactions in skin, we used a human keratin 14 promoter to target expression of human KGF cDNA to the stratified squamous epithelia of transgenic mice. Mice expressing KGF in their epidermis typically appeared frail and weak, and often had grossly wrinkled skin. These mice exhibited a gross increase in epidermal thickness accompanied by alterations in epidermal growth and differentiation. Most remarkably, animals displayed several striking and unexpected changes, including a marked suppression of hair follicle morphogenesis and suppression of adipogenesis. With age, some animals developed gross transformations in the tongue epithelium and in epidermis. In addition, they exhibited elevated salivation and their salivary glands showed signs of altered differentiation. Collectively, our findings provide new and important insights into the roles of KGF, implicating this potent growth factor in eliciting global effects not only on growth, but also on development and differentiation, of skin and other tissues. In particular, KGF seems to interfere with signalling of some mesenchymal-epithelial interactions.
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Glia-activating factor (GAF) is a novel heparin-binding growth factor purified from the culture supernatant of a human glioma cell line. It shows a spectrum of activity slightly different from those of other known growth factors. We have isolated the cDNA which encodes human GAF. A homology search revealed that GAF would be the ninth member of the FGF family, and we therefore call it FGF-9. The human FGF-9 cDNA cloned by using oligonucleotide probes encoded a polypeptide consisting of 208 amino acids. Sequence similarity to other members of the FGF family was estimated to be around 30%. Two cysteine residues and other consensus sequences in family members were also well conserved in the FGF-9 sequence. FGF-9 was found to have no typical signal sequence in its N terminus like those in acidic FGF and basic FGF. Acidic FGF and basic FGF are known not to be secreted from cells in a conventional manner. However, FGF-9 was found to be secreted from cells after synthesis despite its lack of a typical signal sequence. It could be detected exclusively in the culture medium of cDNA-transfected COS cells. The amino acid sequence of proteins purified from culture supernatant of the CHO cell line, which was cDNA transfected and selected as a high producer of FGF-9, showed that no peptides were cleaved from the N terminus except the initiation methionine. The rat FGF-9 cDNA was also cloned, and the structural analysis indicated that the PGF-9 gene is highly conserved. Expression of the FGF-9 gene could be detected in the brain and kidney of the adult rat. Restricted gene expression in organs and the unique secretion nature of the protein suggest that FGF-9 plays a physiological role which differs from those of well-characterized acidic FGF and basic FGF.
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We isolated the cDNA encoding a novel member of the fibroblast growth factor (FGF) family from rat embryos by homology-based polymerase chain reaction. The FGF-related cDNA encodes a protein of 215 amino acids (∼24 kDa), which has a conserved ∼120-amino acid core with ∼30-60% amino acid sequence identity with the FGF family. This protein with a hydrophobic amino terminus appears to be a secreted protein. The cDNA was translated in a coupled in vitro transcription-translation system. The molecular mass of the translation product was observed to be ∼26 kDa. The expression of the FGF-related mRNA in the rat embryo and adult tissues was determined by Northern analysis and in situ hybridization. The mRNA was expressed in several discrete regions of the embryo. In adult tissues, the mRNA was preferentially expressed in the lung. The expression profile of the FGF-related mRNA was different from those of other FGF family mRNAs. As this protein is the 10th documented protein related to FGFs, we tentatively term this protein FGF-10.
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Fibroblast growth factors (FGFs) are essential molecules for mammalian development. The nine known FGF ligands and the four signaling FGF receptors (and their alternatively spliced variants) are expressed in specific spatial and temporal patterns. The activity of this signaling pathway is regulated by ligand binding specificity, heparan sulfate proteoglycans, and the differential signaling capacity of individual FGF receptors. To determine potentially relevant ligand-receptor pairs we have engineered mitogenically responsive cell lines expressing the major splice variants of all the known FGF receptors. We have assayed the mitogenic activity of the nine known FGF ligands on these cell lines. These studies demonstrate that FGF 1 is the only FGF that can activate all FGF receptor splice variants. Using FGF 1 as an internal standard we have determined the relative activity of all the other members of the FGF family. These data should serve as a biochemical foundation for determining developmental, physiological, and pathophysiological processes that involve FGF signaling pathways.
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Pbx1 is a homeodomain transcription factor that has the ability to form heterodimers with homeodomain proteins encoded by the homeotic selector (Hox) gene complexes and increase their DNA-binding affinity and specificity. A current hypothesis proposes that interactions with Pbx1 are necessary for Hox proteins to regulate downstream target genes that in turn control growth, differentiation and morphogenesis during development. In pre B cell leukemias containing the t(1;19) chromosome translocation, Pbx1 is converted into a strong transactivator by fusion to the activation domain of the bHLH transcription factor E2A. The E2A-Pbx1 fusion protein should therefore activate transcription of genes normally regulated by Pbx1. We have used the subtractive process of representational difference analysis to identify targets of E2A-Pbx1. We show that E2A-Pbx1 can directly activate transcription of a novel member of the fibroblast growth factor family of intercellular signalling molecules, FGF-15. The FGF-15 gene is expressed in a regionally restricted pattern in the developing nervous system, suggesting that FGF-15 may play an important role in regulating cell division and patterning within specific regions of the embryonic brain, spinal cord and sensory organs.
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We isolated the cDNA encoding a novel member (207 amino acids) of the fibroblast growth factor (FGF) family from rat embryos. Because this protein is the 18th documented member of the FGF family, we tentatively termed it FGF-18. We have also determined mouse and human FGF-18 with high amino acid identity (99.5 and 99.0%) to rat FGF-18, respectively. Among FGF family members, FGF-18 is most similar (52.7% amino acid identity) to FGF-8 and FGF-17. FGF-18 has a typical signal sequence at its amino terminus. Recombinant rat FGF-18, which was efficiently secreted by High Five insect cells infected with recombinant baculovirus containing the cDNA, induced neurite outgrowth in PC12 cells. The expression of FGF-18 mRNA was examined in adult rat tissues and embryos by Northern blotting analysis and in situ hybridization. FGF-18mRNA of ∼2.7 kilobases was preferentially detected in the lung among adult rat tissues examined. In rat embryos, FGF-18mRNA was detected in several discrete regions at embryonic days 14.5 and 19.5 but not at E10.5. The temporal and spatial patterns ofFGF-18 mRNA expression in embryos are quite different from those of FGF-8 and FGF-17 mRNAs reported. The present results indicate that FGF-18 is a unique secreted signaling molecule in the adult lung and developing tissues.
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