Publications (10)10.67 Total impact
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Article: Artificial gynogenesis and assessment of homozygosity in meiotic gynogens of spotted halibut (Verasper variegatus)
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ABSTRACT: In the present study, methodology of gynogenetic induction in spotted halibut were developed and optimized; the sex ratio of putative meiotic gynogenetic diploids was determined using AFLP-based molecular sexing technique; the homozygosity of gynogenetic population was assessed as opposed to cultivated population. The results showed that high percentage of meiotic gynogenetic diploids were generated when the eggs fertilized with irradiated heterologous sea perch frozen sperm (30–50mJcm−2) were cold shocked in sea water of −1°C for 40–75min at 5min after fertilization. About 15,200 diploid gynogenetic larvae were achieved and they exhibited normal morphology similar to diploid control. The gynogenetic diploids were 100% female, which first confirmed the female homogamete (XX/XY) sex determination in spotted halibut. The genetic analysis showed that the average H O was, respectively, 0.404 and 0.724 in gynogenetic population and cultivated population, indicating an increase of homozygosity in gynogenetic population. Keywords Verasper variegatus -Gynogenesis-Microsatellite-HomozygosityAquaculture International 04/2012; 18(6):1151-1161. · 0.91 Impact Factor -
Article: Growth differences and dimorphic expression of growth hormone (GH) in female and male Cynoglossus semilaevis after male sexual maturation.
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ABSTRACT: Half-smooth tongue sole, Cynoglossus semilaevis, is an ideal model to investigate the regulatory mechanisms of sexual growth dimorphism in fish species. The aim of the study was to investigate the effect of differential age of sexual maturity for females and males on growth and GH mRNA expression in C. semilaevis. The body weight differences between the sexes were not significant in C. semilaevis at age 5 months when females and males were all immature. Significant differences in body weight between the sexes were found after early sexual maturation of males at the age of 9 months. The body weight of 21-month-old females (621.4 ± 86.4g), still not immature, was even 3.28 times higher than that of the males (189.7 ± 14.4g). The cDNAs encoding GH in C. semilaevis was cloned. The GH gene is 2924bp long and consists of six exons and five introns. The results of qRT-PCR showed that GH mRNA levels of the immature females were not significantly different from that of immature males at age 5 months. However, GH mRNA levels of the immature females were significantly higher compared with those of the mature males at age 9 months (P<0.05). At age 11 months, GH mRNA levels of females were even 6.4-fold higher than that of males. In conclusion, for the first time we show that early sexual maturity of males is the main cause of sexual growth dimorphism in C. semilaevis and exert significant effect on GH mRNA expression.Marine Genomics 03/2011; 4(1):9-16. · 1.55 Impact Factor -
Article: Growth differences and differential expression analysis of pituitary adenylate cyclase activating polypeptide (PACAP) and growth hormone-releasing hormone (GHRH) between the sexes in half-smooth tongue sole Cynoglossus semilaevis.
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ABSTRACT: Pituitary adenylate cyclase activating polypeptide (PACAP) and growth hormone-releasing hormone (GHRH) are regulators of growth hormone secretion. In this article, we examined the difference in growth and mRNA expression of PACAP and GHRH between the sexes in half-smooth tongue sole, an important cultured fish species indicating sexually growth dimorphism in China. Firstly, a significant body weight difference between females and males was first observed at 7 months (P<0.05) and at 18 onths the mean body weight of the females (771.0±44.3 g) was as much as 4.9 times higher than that of males (130.6±6.0 g). As a result, half-smooth tongue sole, Cynoglossus semilaevis, is a good model to investigate the effects of growth-related genes expression on sexual growth dimorphism. Secondly, the cDNAs encoding PRP/PACAP and GHRH were isolated. Two differently processed mRNA transcripts of PRP/PACAP (PRP-encoding and PRP splice variant) were found. PACAP and GHRH mRNA was highly abundant in brain and less abundant in other tissues. However, PACAP mRNA was expressed in most brain regions, and was lower in the cerebellum. GHRH mRNA was predominantly expressed in the hypothalamus and weakly expressed in all areas of the brain examined. Ontogenetic expression analysis indicated that PACAP and GHRH mRNA was detected in the early stages of embryogenesis. Finally, differential expression showed that there was no significant difference of the expression level of PACAP or GHRH between the sexes before 8 months of age. However, between 9 and 12 months of age, the GHRH mRNA expression level in males was significantly higher than in females (P<0.05), which might be associated with GH deficiency in males. In contrast, the male PACAP mRNA expression level was not significantly higher than that in females even at 9 and 12 months of age. The present results provide important clues for understanding the sexual growth dimorphism mechanisms in half-smooth tongue sole.General and Comparative Endocrinology 01/2011; 170(1):99-109. · 3.27 Impact Factor -
Article: Artificial gynogenesis in Cynoglossus semilaevis with homologous sperm and its verification using microsatellite markers
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ABSTRACT: Effective methods for induction of gynogenetic diploids in Cynoglossus semilaevis are needed to initiate monosex culture. An effective protocol to induce half-smooth tongue sole gynogenesis using homologous sperm was developed in this study. A UV dose of 50 mJ cm−2 was found to be the most effective for genetic inactivation of tongue sole sperm. Treatment optima for cold shocks were 5 °C for 20–23 min at 5 min after fertilization and the hatching rate of gynogenetic diploids was 10.0%. Microsatellite analysis at locus Csou 6 revealed that there was no genetic contribution from the paternal genome in 24 progenies of a meiotic gynogenetic family. Polymerase chain reaction demonstrated that only four individuals of 24 meiotic gynogenetic diploids produced the female-specific band of about 205 bp. The female/male ratio of gynogenetic diploids was significantly different from the theoretical ratio of 1:1. It is possible that there are some recessive lethal genes in W chromosome.Aquaculture Research 10/2009; 41(6):913 - 920. · 1.20 Impact Factor -
Article: Isolation and characterization of 19 EST‐linked ploymorphic microsatellite loci for olive flounder (Paralichthys olivaceus)
Aquaculture Research 04/2009; 40(8):980 - 983. · 1.20 Impact Factor -
Article: Production of cloned goats by nuclear transfer of cumulus cells and long-term cultured fetal fibroblast cells into abattoir-derived oocytes.
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ABSTRACT: Dairy goats are ideal for the transgenic production of therapeutic recombinant proteins. The use of recombinant somatic cell lines for nuclear transfer (NT) allows the introduction of genes by transfection, increases the efficiency of transgenic animal production to 100%, and overcomes the problem of founder mosaicism. Although viable animals have been cloned via NT from somatic cells of 11 species, the efficiency has been extremely low. Both blastomere and somatic cell NT increased fetal loss and perinatal morbidity/mortality in cattle and sheep, but fetal loss and perinatal mortality appear to be relatively low in goats. In this study, we produced cloned goats by NT from cumulus cells and long-term cultured fetal fibroblast cells (FFCs) to abattoir-derived oocytes. NT embryos were constructed from electrofusion of cumulus cells (CCs), FFCs, or skin fibroblast cells (SFCs) with cytoplasts prepared from abattoir-derived ovaries. The NT embryos were activated with an optimized activating protocol (1 min exposure to 2.5 microM ionomycin followed by 2 hr incubation in 2mM 6-DMAP). Two viable cloned kids from CCs and one from long-term cultured FFCs (at passage 20-25) were born. Microsatellite analysis of 10 markers confirmed that all cloned offspring were derived from corresponding donor cells. To our knowledge, the production of cloned goat offspring using abattoir-derived oocytes receiving nuclei from CCs and long-term cultured FFCs has not been reported. The production of viable cloned animals after activation with reduced intensity of ionomycin and 6-DMAP treatment has also not been reported. Loss of cloned embryos was obvious after 45 and 90 days of pregnancy, and a lack of cotyledons, heart defects, and improperly closed abdominal wall were observed in the aborted fetuses and one cloned kid. The fusibility and in vitro developmental potential of embryos reconstructed from FFCs at passage 20-25 were significantly lower than those of embryos reconstructed from FFCs at passage 3-5, and the cloning efficiency of the long-term cultured cells was low (0.5%).Molecular Reproduction and Development 08/2006; 73(7):834-40. · 2.53 Impact Factor -
Article: [Identification and analysis of a novel microsatellite marker flanking porcine myostatin gene (MSTN)].
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ABSTRACT: In animal breeding, microsatellite marker plays an important role in constructing genetic maps, QTL mapping and function analysis of structural genes. Myostatin, also known as GDF8, is a negative regulator of skeletal muscle mass and, in swine, it is evidenced to be related to birth weight and average daily gain from 60 kg to 100 kg of body weight. In present study, by subcloning and sequencing,we identified a novel microsatellite marker which is useful for fine QTL mapping for meat traits. A BAC clone containing porcine MSTN was extracted and digested with EcoR I to recover the fragment of > 4 kb for subcloning in pGEM-3zf (+). Sequencing and alignment results showed that this subcloned fragment was not from porcine MSTN, but included a tandem repeat of (TG) 13, which is a novel microsatellite marker (GenBank accession number: AF454400) flanking MSTN. To exclude its vector origin we designed specific primers flanking this marker and successfully amplified this fragment from porcine genome. Through a pedigree analysis of a double-muscled Yorshire strain, we found that it is inherited in a co-dominant manner. We also checked the gene frequencies of this locus in 381 unrelated individuals of 7 pig breeds, namely Laiwu,Landrace, Yorkshire,Duroc, Peterian, Min and Erhualian. Only two alleles were detected, the repeating number of which are 13 (allele A) and 19 (allele B) respectively, which indicated that it is a low poly morphic microsatellite marker. In addition, the frequencies of the two alleles are different between the two types of pig breeds, while allele A is dominant in Chinese local breeds, allele B is dominant in imported breeds. Alignment with AY208121 indicate that this locus is located 42 kb downstream of porcine MSTN. We speculate that this microsatellite DNA is an important marker both in fine QTL mapping for meat traits and in the expression study of porcine MSTN.Acta Genetica Sinica 06/2004; 31(5):480-4. -
Article: [Studies of BMPR-IB and BMP15 as candidate genes for fecundity in little tailed han sheep].
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ABSTRACT: BMPR-IB gene which controls the fecundity of Booroola Merino and BMP15 gene which affects the ovulation of Invedale and Hanna were studied as candidate genes on the fecundity of Little Tailed Han Sheep, and their mutations and genetic effects were analyzed. The results showed that there was a same mutation in BMPR-IB gene (A746G) of Little Tailed Han Sheep as that of Booroola Merino. The BB mutation genotype was superior in prolific Little Tailed Han Sheep, and the ewes with genotype BB had 0.97(P < 0.05) and 1.5(P < 0.01) lambs more than those with genotype + + in the first parity and later parities, respectively. It could be inferred that BMPR-IB gene was related with the major gene that controls the high prolificacy of Little Tailed Han Sheep. While there was not mutation of V31D or Q23Ter in BMP15 gene of Little Tailed Han Sheep, it showed that the fecundity mechanism of Little Tailed Han Sheep was different from that of Romney sheep. Then it was ruled out the possibility that the ovulation of Little Tailed Han Sheep was affected by the mutation of BMP15 gene.Acta Genetica Sinica 09/2003; 30(8):755-60. -
Article: [Relationship of T-->A mutation in the promoter region of myostatin gene with growth traits in swine].
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ABSTRACT: A T-->A mutation in the promoter region of porcine myostatin (MSTN) gene has been identified in previous work. Associations of the myostatin genotypes with growth traits are unknown in swine. The present study attempts to analyze the relationship of the mutation with the growth traits which included body weight at 60 d (BW60), average daily gain from 25 kg to 60 kg(ADG1), average daily gain from 60 kg to 100 kg (ADG2) and average daily gain from 25 kg to 100 kg (ADG). Data from 165, 275, 276 and 276 unrelated individuals respectively were collected from three different swine breeding companies. Detections of the mutation were carried out by PCR-RFLP approach. The effect of MSTN genotypes (TT and TA) on growth traits was estimated by GLM procedure. The results showed that for ADG2, individuals with TA genotype were higher than those of TT genotype (P = 0.052), indicating a positive effect for A allele. For BW60, ADG1 and ADG, the effect of porcine MSTN genotype was non-significant (P > 0.1). Studies are still necessary for examining the effects in "double-muscled" pigs.Acta Genetica Sinica 05/2002; 29(5):413-6. -
Article: [Research on Alu element inserted mutation in porcine FSH beta subunit gene].
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ABSTRACT: The inserted fragment of FSH beta subunit gene in Laiwu pigs, an excellent local pig breed in North China, Duli pigs and Landrace pigs was amplified, cloned and sequenced. Sequence analysis showed that the length of the fragments inserted between +809 bp and +810 bp of the published sequences (D00621) were 275 bp, 277 bp and 274 bp in Laiwu, Duli and Landrace pigs, respectively. And the poly (A)s in these inserted fragments were 17, 19 and 16 adenines, respectively. They were all shorter than those in Taihu pigs(292 bp and 32 adenines, respectively) reported formerly. According to the RNA polymerase III promoter structure and Alu I restriction enzyme site in the inserted fragment, it should be regarded as an Alu element. FSH beta subunit gene was considered as one candidate gene of pig litter size trait, while RNA polymerase III promoter could promote the transcription of the neighboring chromosome sequence so as to control the expression of FSH beta subunit gene or other genes. As a result, the major difference of the inserted fragment among different pig breeds was originated from the length of poly (A) end. So it was assumed that the poly (A) structure in the inserted fragment could influence the pig litter size. Because Laiwu pigs with genotype AA had 1.2 litter size more than those with genotype BB, it could be concluded that FSH beta subunit gene was related with pig litter size traits or linked with the genes of which, and the poly(A) structure in this Alu element played a critical role.Acta Genetica Sinica 02/2002; 29(11):977-82.
Top co-authors
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Institutions
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2002–2012
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Shandong Agricultural University
- College of Animal Science and Technology
China
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2002–2004
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China Agricultural University
- State Key Laboratory for Agrobiotechnology
Beijing, Beijing Shi, China
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