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ABSTRACT: The inhibin/activin subunits (α, βA and βB) have been found in epididymal tissue of many mammals, but there have been no data available for wild seasonal breeders so far. The aim of this study was to investigate the immunoreactivities of inhibin/activin α, βA and βB subunits in the epididymis of wild ground squirrels during the breeding and nonbreeding seasons. Immunohistochemistry and Western blotting were performed to detect the epididymal immunolocalizations and immunoreactivities of the three subunits. Strong immunostaining of α subunit was present in the interstitial part of the caput epididymis and epithelial parts of the corpus epididymis and cauda epididymis during the breeding season, whereas no α subunit was found in the nonbreeding season. βA and βB subunits were expressed in all cell types of the epithelium throughout the whole seasonal cycle, and immunostaining in the breeding season was likely stronger compared with that of the nonbreeding season. These results suggested that the epididymis might be a potential source of inhibin and activin in the wild male ground squirrel, and the secretion of epididymal inhibin and activin showed distinct seasonal changes. Furthermore, inhibin and activin might function as paracrine and/or autocrine factors that have an effect on the epididymis.
Journal of Reproduction and Development 03/2013; · 1.46 Impact Factor
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ABSTRACT: The PBAN (pheromone biosynthesis activating neuropeptide)/pyrokinin peptides comprise a major neuropeptide family characterized by a common FXPRL amide at the C-terminus. These peptides are actively involved in many essential endocrine functions. For the first time, we reported the cDNA cloning and sequence determination of the PBAN from the seabuckthorn carpenterworm, Holcocerus hippophaecolus, by using rapid amplification of cDNA ends. The full-length cDNA of Hh-DH-PBAN contained five peptides: diapause hormone (DH) homolog, α-neuropeptide (NP), β-NP, PBAN, and γ-NP. All of the peptides were amidated at their C-terminus and shared a conserved motif, FXPR (or K) L. Moreover, Hh-DH-PBAN had high homology to the other members of the PBAN peptide family: 56% with Manduca sexta, 66% with Bombyx mori, 77% with Helicoverpa zea, and 47% with Plutella xylostella. Phylogenetic analysis revealed that Hh-DH-PBAN was closely related to PBANs from Noctuidae, demonstrated by the relatively higher similarity compared with H. zea. In addition, real-time quantitative PCR (qRT-PCR) analysis showed that Hh-DH-PBAN mRNA expression peaked in the brain-subesophageal ganglion (Br-SOG) complex, and was also detected at high levels during larval and adult stages. The expression decreased significantly after pupation. These results provided information concerning molecular structure characteristics of Hh-DH-PBAN, whose expression profile suggested that the Hh-DH-PBAN gene might be correlated with larval development and sex pheromone biosynthesis in females of the H. hippophaecolus.
Archives of Insect Biochemistry and Physiology 01/2013; · 1.36 Impact Factor
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ABSTRACT: The aim of this study was to elucidate the regulatory role of androgen in the follicular development of wild female ground squirrels. Immunohistochemical staining of FSHR, LHR, P450c17, P450arom, androgen receptor (AR), estrogen receptors (ERa and ERb) were executed in ovaries of female ground squirrels from both breeding and nonbreeding seasons. In addition, total ovarian proteins were extracted from the ovaries of squirrels from breeding and nonbreeding seasons, and Western blot analysis were performed in order to probe for FSHR, LHR, P450c17, P450arom, AR, ERa and ERb. The results of immunohistochemical staining and Western blotting of P450c17 showed that there was no significant difference between the breeding and nonbreeding seasons. It was found that granulosa cells expressed P450arom during the breeding season. In contrast, there was no positive staining of P450arom in the nonbreeding season. There was no significant difference in immunoreactivity of AR between the breeding and nonbreeding seasons. However, the immunoreactivities of ERa and ERb were both significantly reduced in the nonbreeding season compared to the breeding season. The positive stains of FSHR and LHR were found in the granulosa cells and theca cells of the ovaries of the breeding and nonbreeding seasons. In addition, the Western blotting results of FSHR and LHR showed a significant reduction in the nonbreeding season compared with the breeding season. These findings suggested that androgen might be predominantly converted into estrogen in order to regulate the follicular development via binding of estrogen receptors during the breeding season, whereas androgen might predominantly directly bind androgen receptor to regulate the follicular development during the nonbreeding season in the ovaries of wild female ground squirrels.
Journal of Ovarian Research 09/2012; 5(1):26. · 2.57 Impact Factor
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ABSTRACT: The intraovarian function of gonadally produced inhibin and activin has been extensively studied in experimental models for decades, yet their presence and function have been rarely reported in wild rodents. With our seasonal breeding model, the wild ground squirrel, we aimed to investigate the possible roles of these peptides in the seasonal folliculogenesis. Immunohistochemical staining and Western blot have been used to detect the cellular localization and expression patterns of inhibin/activin subunits (α, β(A) and β(B)). In the breeding season ovary, all three subunits were present in granulosa cells, theca cells of antral follicles and interstitial cells, with the strongest immunostaining in granulosa cells. Following ovulation, the corpora lutea become a major site of inhibin/activin synthesis. In the nonbreeding season ovary, inhibin/activin α and β(A) subunits were weakly immunopositive in granulosa cells of early stage follicles, while β(B )subunit was undetectable. The expression level of inhibin/activin subunit proteins were generally higher in the ovaries of the breeding season, and then decreased to a relatively low level during the nonbreeding season. The dynamic expression of inhibin/activin subunits indicated that they might play important paracrine and/or autocrine role(s) during the seasonal folliculogenesis of the wild ground squirrel.
Journal of Reproduction and Development 06/2012; · 1.46 Impact Factor
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ABSTRACT: In this study, we investigated the immunoreactivity of vascular endothelial growth factor (VEGF) and its receptors flt-1 (VEGFR1) and the kinase domain receptor (KDR/Flk-1, VEGFR2) in the uteri of the wild ground squirrels during the estrous period, early pregnancy and nonbreeding period. Cellular localizations of VEGF, VEGFR1 and VEGFR2 were detected by immunohistochemistry, and total proteins were extracted from uterine tissue in the estrous period, early pregnancy and nonbreeding period for Western blotting analysis. In addition, plasma estradiol-17β and progesterone concentrations were measured by radioimmunoassay. Stronger positive staining of VEGF was found in luminal epithelial cells and glandular cells, and its receptors (VEGFR1 and VEGFR2) were observed in stromal cells in the estrous period and early pregnancy compared with the nonbreeding period. The protein levels of VEGF, VEGFR1 and VEGFR2 were significantly higher in the estrous period and early pregnancy as compared with the nonbreeding period. Besides, plasma estradiol-17β and progesterone concentrations were higher in the estrous period and early pregnancy than in the nonbreeding period, suggesting that the immunoreactivities of VEGF, VEGFR1 and VEGFR2 were correlated with changes in plasma estradiol-17β and progesterone concentrations. These results suggested that VEGF and its receptors may be involved in the regulation of seasonal changes in the uterine functions of wild female ground squirrels.
Journal of Reproduction and Development 06/2012; · 1.46 Impact Factor
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Nature 04/2012; 484(7395):455. · 36.28 Impact Factor
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ABSTRACT: The platelet-derived growth factor (PDGF) system is expressed and can exert its biological role in the male reproductive system including the maintenance of morphological structure and function of the epididymis. The aim of this study was to clarify the relationship between the PDGF system and seasonal changes in morphology of the wild ground squirrel epididymis during the breeding and nonbreeding seasons. Hematoxylin-eosin (HE) staining was used to observe the epididymal morphology and histology. Immunohistochemistry and Western blotting were performed to detect the immunoreactivities of PDGF-A and B and PDGFR-α. Significant seasonal changes in epididymal morphology were observed in the breeding and nonbreeding seasons. The proportions of the three compartments (interstitial tissue, epithelium and lumen of the duct) revealed distinct variances. Strong immunostaining of PDGF-A was present in the myoid cell and on the sperm in the breeding season, whereas there was a faint signal in the myoid cell in the nonbreeding season. PDGFR-α was expressed in all cell types of the epithelium throughout the whole seasonal cycle, and immunostaining of PDGFR-α in the breeding season was significantly stronger compared with that of the nonbreeding season. PDGF-B was not detected in the epididymis of wild ground squirrels. These results suggested that seasonal morphological changes in epididymis were correlated with immunoreactivities of PDGF-A and its receptor PDGFR-α and that PDGF-A and PDGFR-α might function as paracrine, autocrine or apocrine factors in wild ground squirrels.
Journal of Reproduction and Development 03/2012; 58(3):353-9. · 1.46 Impact Factor
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Ben Li,
Xia Sheng,
Moshi Song, Haolin Zhang,
Jiaju Weng,
Mengyuan Zhang,
Xiao Hu,
Jiao Zhou,
Meiyu Xu,
Qiang Weng,
Gen Watanabe,
Kazuyoshi Taya
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ABSTRACT: In this study, we investigated the presence of nerve growth factor (NGF) and its receptors tyrosine kinase A (TrkA) and p75 in the uterus of the wild ground squirrels during the estrous period, early pregnancy and non-breeding period. In the estrous period and early pregnancy, NGF and TrkA were immunolocalized in stromal cells, luminal epithelial cells, glandular cells and smooth muscle cells whereas in the non-breeding period, both of them were detected only in luminal epithelial cells and glandular cells, but not in stromal cells or smooth muscle cells. Stronger immunostaining of NGF and TrkA was observed in luminal epithelial cells and glandular cells in the estrous period and early pregnancy as compared to the non-breeding period. p75 was immunolocalized only in luminal epithelial and glandular cells during the estrous period, early pregnancy and non-breeding period. The intensity of the immunohistochemical signals for p75 did not vary significantly in the estrous period, early pregnancy and non-breeding period. The mean mRNA levels of NGF and TrkA and p75 were significantly higher in the estrous period and early pregnancy as compared to the non-breeding period. Besides, plasma estradiol-17β and progesterone concentrations were higher in the estrous period and early pregnancy than in the non-breeding period, suggesting that the expression patterns of NGF and TrkA are correlated with changes in plasma estradiol-17β and progesterone concentrations. These results indicate that NGF and its receptor TrkA may be involved in the regulation of seasonal changes in the uterine functions of wild female ground squirrels.
General and Comparative Endocrinology 03/2012; 176(1):62-9. · 3.27 Impact Factor
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Xia Sheng, Haolin Zhang,
Mengyuan Zhang,
Wei Zhang,
Xiao Hu,
Moshi Song,
Jiao Zhou,
Meiyu Xu,
Qiang Weng,
Gen Watanabe,
Kazuyoshi Taya
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ABSTRACT: The seasonal spermatogenesis and localization of inhibin/activin subunits (alpha, betaA, betaB) in the testes of wild ground squirrel has been previously described; however, the expression pattern of activin receptors and cytoplasmic signaling SMADs has not been detected in any seasonal breeders. The objective of this study was to investigate the abundance and cellular localization of activin signaling components in testes of the wild ground squirrel during the breeding and nonbreeding seasons. The immunolocalizations of ActRIIB (activin type II receptor B) and activin-related SMADs (phospho-SMAD2/3, SMAD4 and SMAD7) were observed by immunohistochemistry. Total proteins were extracted from testicular tissues in the breeding and nonbreeding seasons and were used for Western blotting analysis for ActRIIB and SMADs. Immunoreactivities of activin signaling components were greater in the testes of the breeding season, and then decreased to a relatively low level in the nonbreeding season. ActRIIB and related SMADs were widely spread in the active testes, while spermatogonia were the predominant cellular sites of activin signal transduction during arrested spermatogenesis. The dynamic regulation of activin type II receptor and SMADs indicated that the activin signal pathway played an important paracrine role in seasonal spermatogenesis of the wild ground squirrel. Furthermore, the distinct localizations and immunoreactivity of ActRIIB and SMADs might suggest different functions of activin in seasonal spermatogenesis.
Journal of Reproduction and Development 11/2011; 58(1):126-31. · 1.46 Impact Factor
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Lu Lu, Haolin Zhang,
Na Lv,
Xiaoting Ma,
Long Tian,
Xiao Hu,
Shuqiang Liu,
Meiyu Xu,
Qiang Weng,
Gen Watanabe,
Kazuyoshi Taya
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ABSTRACT: Aromatase cytochrome P450 (P450arom) is an enzyme that catalyzes the conversion of androgen to estrogen. Expression of P450arom in extra-gonadal sites and locally-synthesized estrogen play an important role in physiological conditions. The purpose of this study was to investigate the cellular immunolocalization of androgen receptor (AR), P450arom, estrogen receptor alpha (ERa) and estrogen receptor beta (ERβ) in muskrat scent glands during the breeding season. Histological observation and immunohistochemistry of AR, P450arom, ERa and ERβ were performed in the muskrat scent glands. In addition, total proteins were extracted from scent glandular tissues in the breeding season and were used for Western blotting analysis for AR, P450arom, ERα and ERβ. Histologically, glandular cells, interstitial cells, epithelial cells of the excretory duct and the excretory tubules were identified in the muskrat scent glands during the breeding season. AR was only observed in glandular cells of scent glands; P450arom was expressed in glandular cells and epithelial cells of the excretory duct; ERα was found in glandular cells, interstitial cells and epithelial cells of the excretory duct, whereas ERβ was present in glandular cells and epithelial cells of the excretory duct. Also, the positive signals of AR, P450arom, ERα and ERβ by Western blotting were all observed in scent glandular tissues. These results suggested that the scent gland is the target organ of androgens and estrogens, and that estrogens may play an important autocrine or paracrine role in glandular function of the muskrats.
ZOOLOGICAL SCIENCE 10/2011; 28(10):727-32. · 0.95 Impact Factor
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Yong Shen,
Yuning Liu,
Jiajia Ma,
Xiaoting Ma,
Yang Tian, Haolin Zhang,
Lin Li,
Meiyu Xu,
Qiang Weng,
Gen Watanabe,
Kazuyoshi Taya
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ABSTRACT: The objective of this study was to investigate immunoreactivity of the c-kit receptor in the oviduct of Rana chensinensis during the prehibernation period. Histological examination of oviducts was performed during the prehibernation period. The sections of oviduct were immunostained by the avidin-biotin-peroxidase complex method using rabbit polyclonal antisera raised against the rat c-kit receptor and PCNA. Total proteins were extracted from oviducal tissues and used for Western blotting analysis. Immunohistochemistry revealed the presence of the c-kit receptor and PCNA in the oviduct tissues during the prehibernation period. Also, positive signals for the c-kit receptor and PCNA by Western blotting were observed in oviduct tissues during the prehibernation period. These results suggested that the c-kit receptor might play a regulatory role in oviducal hypertrophy in the brown frog, Rana chensinensis.
Journal of Veterinary Medical Science 09/2011; 74(2):209-13. · 0.85 Impact Factor
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Journal of Neuroscience 09/2011; 31(38):13343-5. · 7.11 Impact Factor
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ABSTRACT: Acupuncture and electro-acupuncture (EA) are now widely used to treat disorders like pain. We and others have shown previously that current frequency, intensity and treatment duration all significantly influence the anti-nociceptive effects of EA. There is evidence that stimulating sites also affect the antinociception, with EA applied ipsilaterally to the pain site being more effective under some pain states but contralateral EA under others. It was recently reported that local adenosine A1 receptors were responsible for ipsilateral acupuncture, but what mechanisms specifically mediate the anti-nociceptive effects of contralateral acupuncture or EA remains unclear. In the present study, we applied 100 Hz EA on the ipsi- or contra-lateral side of rats with inflammatory pain induced by intra-plantar injection of formalin, and reported distinct anti-nociceptive effects and mechanisms between them. Both ipsi- and contra-lateral EA reduced the paw lifting time in the second phase of the formalin test and attenuated formalin-induced conditioned place aversion. Contralateral EA had an additional effect of reducing paw licking time, suggesting a supraspinal mechanism. Lesions of rostral anterior cingulate cortex (ACC) completely abolished the anti-nociceptive effects of contra- but not ipsi-lateral EA. These findings were not lateralized effects, since injection of formalin into the left or right hind paws produced similar results. Overall, these results demonstrated distinct anti-nociceptive effects and mechanisms between different stimulating sides and implied the necessity of finding the best stimulating protocols for different pain states.
Molecular Pain 08/2011; 7:61. · 3.53 Impact Factor
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Xiaoting Ma, Haolin Zhang,
Jiaju Weng,
Xia Sheng,
Lu Lu,
Xiao Hu,
Shuqiang Liu,
Meiyu Xu,
Qiang Weng,
Gen Watanabe,
Kazuyoshi Taya
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ABSTRACT: The objective of this study was to investigate the cellular immunolocalization of inhibin a and inhibin/activin (β(A) and β(B)) subunits in the muskrat testes and scented glands during the breeding season. Inhibin α and inhibin/activin (β(A) and β(B)) subunits were expressed in Sertoli cells and Leydig cells of testes and glandular cells of scented glands, respectively. Also, positive signals of inhibin α and inhibin/activin (β(A) and β(B)) subunits by Western blotting were both observed in testicular and scented glandular tissues. These results suggested that the testes and scented glands of the muskrats had the ability to synthesize inhibins and activins and that activins and inhibins might play an important role in testicular and scented glandular function in muskrats.
Journal of Veterinary Medical Science 05/2011; 73(9):1199-205. · 0.85 Impact Factor
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Haolin Zhang,
Xia Sheng,
Xiao Hu,
Xiuwen Li,
Hui Xu,
Mengyuan Zhang,
Ben Li,
Meiyu Xu,
Qiang Weng,
Zhixiang Zhang,
Kazuyoshi Taya
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ABSTRACT: The purpose of this study was to investigate seasonal changes of spermatogenesis and the cellular localization of P450c17 and P450arom in wild male ground squirrels during the breeding and non-breeding seasons. The testicular weight, testicular size and score count of spermatogenesis from April to September were measured, and histological and immunohistochemical observations of testicular tissues were performed in wild male ground squirrels. In addition, total proteins were extracted from testicular tissue in the breeding and non-breeding seasons and were used for Western blotting analysis for P450c17 and P450arom. There were marked variations in testicular weight, testicular size and score count of spermatogenesis from the breeding season (April) to the non-breeding season (September). Histologically, spermatogonia, primary spermatocytes, secondary spermatocytes and spermatozoa were identified in the breeding season (April). Immunolocalization of P450c17 was detected in Leydig cells and spermatozoa during the breeding season and was only found in Leydig cells during the non-breeding season. The positive signals of P450c17 by Western blotting were both observed in the breeding and non-breeding seasons. Immunolocalization of P450arom was observed in Leydig cells, Sertoli cells and all types of spermatogenic cells including mature-phase spermatozoa in the breeding season, while immunoreactivity for P450arom was not present in the testis of the non-breeding season. With P450arom antibody, a band was also only detected in the breeding season by Western blotting. These results suggest that the seasonal changes in testicular weight and size are correlated with spermatogenesis and immunolocalization of P450c17 and P450arom, and androgen and estrogen may play an important role in the spermatogenesis and testicular recrudescence and regression process.
Journal of Reproduction and Development 03/2010; 56(3):297-302. · 1.46 Impact Factor
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BMJ (Clinical research ed.). 01/2010; 340:c1339.
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ABSTRACT: The ideas about the protective role of fevers in infection are not new.1 The potential benefit of symptoms is not unique to fever and may lead us to a better understanding of “health.”Evidence that inducing a fever could ameliorate the severity of infections was first provided by Jauregg in the 1920s. …
BMJ. 12/2009; 340.
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ABSTRACT: The objective of this study was to investigate the seasonal changes in spermatogenesis and the immunolocalization of the inhibin alpha and inhibin/activin (betaA and betaB) subunits during the breeding and non-breeding seasons in the wild male ground squirrel. The testicular weight and size and seminiferous tubule diameter were measured, and histological observations of testes were performed. The sections of the testes were immunostained by the avidin-biotin-peroxidase complex method (ABC) using polyclonal antisera raised against porcine inhibin alpha, inhibin/activin betaA and inhibin/activin betaB during the breeding and non-breeding seasons. There were marked variations in testicular weight and size and seminiferous tubule diameter between the breeding and non-breeding seasons, and all types of spermatogenic cells, including spermatozoa, were found in the breeding season. In addition, immunoreactivity was also detected for the inhibin alpha, betaA and betaB subunits in Sertoli and Leydig cells during the breeding season, but immunostaining was only present for the inhibin alpha and inhibin/activin betaB subunits in Sertoli cells during the non-breeding season. These results suggest that seasonal changes in testicular weight and size and seminiferous tubule diameter of wild ground squirrels are correlated with changes in spermatogenesis, and the cellular localization of the inhibin/activin subunits showed season related changes in the breeding and non-breeding seasons.
Journal of Reproduction and Development 10/2008; 54(6):460-4. · 1.46 Impact Factor
