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Xiaobing Wu - One of the best experts on this subject based on the ideXlab platform.
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cloning characterisation and expression profile of kisspeptin1 and the kisspeptin1 receptor in the hypothalamic pituitary ovarian axis of chinese Alligator Alligator Sinensis during the reproductive cycle
Reproduction Fertility and Development, 2020Co-Authors: Yongkang Zhou, Ali Izaz, Shulong Duan, Ruidong Zhang, Renping Wang, Xiaobing WuAbstract:Kisspeptin1 (Kiss1), a product of the Kiss1 gene, plays an important role in the regulation of reproduction in vertebrates by activating the Kiss1 receptor (Kiss1R) and its coexpression with gonadotrophin-releasing hormone (GnRH) in GnRH neurons. The purpose of this study was to clone the Kiss1 and Kiss1R genes found in the brain of Alligator Sinensis and to explore their relationship with reproduction. The full-length cDNA of Kiss1 is 816bp, the open reading frame (ORF) is 417bp and the gene encodes a 138-amino acid precursor protein. The full-length cDNA of Kiss1R is 2348bp, the ORF is 1086bp and the gene encodes a 361-amino acid protein. Quantitative polymerase chain reaction showed that, except for Kiss1R expression in the hypothalamus, the expression of Kiss1 and Kiss1Rduring the reproductive period of A. Sinensis was higher than that in the hypothalamus, pituitary gland and ovary during the hibernation period. The changes in GnRH2 mRNA in the hypothalamus were similar to those of GnRH1 and peaked during the reproductive period. This study confirms the existence of Kiss1 and Kiss1R in A. Sinensis and the findings strongly suggest that Kiss1 and Kiss1R may participate in the regulation of GnRH secretion in the hypothalamus of Alligators during the reproductive period. Furthermore, this is the first report of the full-length cDNA sequences of Kiss1 and Kiss1R in reptiles.
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historical population decline and habitat loss in a critically endangered species the chinese Alligator Alligator Sinensis
Global Ecology and Conservation, 2019Co-Authors: Huan Wang, Shulong Duan, Jihui Zhang, Huabin Zhang, Baowei Zhang, Mingsheng Wang, Xiaobing WuAbstract:Abstract Habitat loss and degradation are among the leading causes of local extinctions, so it is crucially important to preserve and restore the remaining critical habitat increasingly critical to conserve biodiversity. However, mechanisms driving species extinction often begin with habitat loss and seldom are well understood, which is greatly limits our ability to mitigate their impacts. The Chinese Alligator ( Alligator Sinensis ) is a critically endangered crocodilian, which is narrowly distributed in very small six regions of Anhui province, China. In the present study, we used 9 nuclear microsatellite loci genotyped across the taxon's distribution area to investigate genetic variation and population demography in A. Sinensis . The Chinese Alligator showed unusually low levels of genetic diversity ( N a, 5.44; N e, 1.97; H e, 0.47) based on 793 individuals. Msvar analyses found recent signals of population decline reflecting a bottleneck (an approximately 9-fold decrease) about 25,000 years ago. Ecological niche modelling has shown that the habitat area has declined (about 8–15 fold) for A. Sinensis when compared with that under different historical climatic conditions. Our results indicated that population decline and habitat loss in critically endangered Chinese Alligators may have acted as intrinsic and extrinsic factors that have impacted the current status of A. Sinensis . In addition, these two unfavorable factors may have also directly contributed to a reduction in the genetic diversity of A. Sinensis .
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molecular cloning of androgen receptor and gene expression of sex steroid hormone receptors in the brain of newborn chinese Alligator Alligator Sinensis
Gene, 2018Co-Authors: Yongkang Zhou, Ali Izaz, Mengjuan Wu, Ruidong Zhang, Renping Wang, Ying Zhang, Xiaobing WuAbstract:Abstract Sex steroid hormones play an important role in mediating physiological responses and developmental processes through their receptors across all vertebrates. Chinese Alligator (Alligator Sinensis) is a critically endangered reptile species unique to China. In this study, we have cloned one of the sex steroid hormone receptor genes, androgen receptor (AR) from the brain of Chinese Alligator for the first time. The full-length AR cDNA is 2717 bp in length with an open reading frame (ORF) encoding 722 amino acids. Amino acid alignment analyses indicated that the ARs exhibit highly conserved functional domains. Especially, the P-box and D-box, which are essential to ensure that receptor binding to the androgen response elements, are completely conserved in selected species. Using the quantitative real-time PCR (qPCR), the spatial expression of four receptor mRNAs in all newborn brain tissues and temporal expression of them in the cerebrum during the embryonic development in Chinese Alligators were investigated. The results of qPCR showed ubiquitous expression of the four receptor mRNAs in all newborn brain tissues examined and significant changes in the expression levels of these receptor mRNAs in the embryonic development. These results suggest that sex steroid hormones might play an important role in the regulation of complex neuroendocrine activities in newborn Chinese Alligator. Furthermore, these data provide an important foundation for further studies on endocrinology and molecular biology of non-mammalian sex steroid hormone receptors.
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insights into the seasonal adaptive mechanisms of chinese Alligators Alligator Sinensis from transcriptomic analyses
Australian Journal of Zoology, 2018Co-Authors: Fang Zhang, Yongkang Zhou, En Li, Rong Wu, Xiaobing WuAbstract:The Chinese Alligator (Alligator Sinensis) is an endemic and rare species in China, and is considered to be one of the most endangered vertebrates in the world. It is known to hibernate, an energy-saving strategy against cold temperatures and food deprivation. Changes in gene expression during hibernation remain largely unknown. To understand these complex seasonal adaptive mechanisms, we performed a comprehensive survey of differential gene expression in heart, skeletal muscle, and kidney of hibernating and active Chinese Alligators using RNA-Sequencing. In total, we identified 4780 genes differentially expressed between the active and hibernating periods. GO and KEGG pathway analysis indicated the likely role of these differentially expressed genes (DEGs). The upregulated DEGs in the active Chinese Alligator, CSRP3, MYG and PCKGC, may maintain heart and skeletal muscle contraction, transport and storage of oxygen, and enhance the body’s metabolism, respectively. The upregulated DEGs in the dormant Chinese Alligator, ADIPO, CIRBP and TMM27, may improve insulin sensitivity and glucose/lipid metabolism, protect cells against harmful effects of cold temperature and hypoxia, regulate amino acid transport and uptake, and stimulate the proliferation of islet cells and the secretion of insulin. These results provide a foundation for understanding the molecular mechanisms of the seasonal adaptation required for hibernation in Chinese Alligators, as well as effective information for other non-model organisms research.
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molecular characterization and tissue expression profiles of insulin like growth factor binding protein 1 igfbp 1 in chinese Alligator Alligator Sinensis during the active and hibernating periods
Biologia, 2018Co-Authors: Mengyun Li, Yongkang Zhou, Shengzhou Zhang, Ting Meng, Fei Peng, Xiaobing WuAbstract:The Chinese Alligator Alligator Sinensis (Fauvel, 1879) is an endangered species endemic to China. Up to this date, very little is known about the regulation of its growth and development. Insulin-like growth factor binding protein-1 (IGFBP-1) plays essential roles in regulating vertebrate growth and development by modulating IGFs action. In this study, we cloned the full-length cDNA of Chinese Alligator IGFBP-1 (caIGFBP-1) for the first time, which has an open reading frame (ORF) of 885-bp nucleotides encoding a peptide of 294 amino acid residues. The caIGFBP-1 contains all features of IGFBP-1 peptide with the conserved cysteine-rich N- and C-terminal domains and variable central L-domain. Multiple alignment analysis showed that caIGFBP-1 shares high identity in amino acid sequence with the IGFBP-1 s of American Alligator Alligator mississippiensis (Daudin, 1802) (99.32%) and birds (66.32–74.53%). Phylogenetic tree analysis of IGFBP-1 amino acid sequences indicated that Alligators cluster into bird branch. The caIGFBP-1 is widely expressed in all the examined tissues, the expression level is highest in liver, higher in kidney, moderate in pancreas, stomach, small intestine, and lower in ovary, muscle, lung, heart. During the hibernating period, the expression level of caIGFBP-1 was significantly reduced in liver, kidney, stomach, pancreas and small intestine, while did not significantly change in heart, lung, ovary and muscle. The caIGFBP-1 mRNA expression changes during the active and hibernating periods, which coincides with that of Chinese Alligator IGF, implicating that it might play important roles in the IGF-mediated growth and development of Chinese Alligator.
Shengguo Fang - One of the best experts on this subject based on the ideXlab platform.
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mhc class i diversity predicts non random mating in chinese Alligators Alligator Sinensis
Heredity, 2019Co-Authors: Haiqiong Yang, Zhenwei Wang, Shengguo FangAbstract:The major histocompatibility complex (MHC) has several important roles in kin recognition, pathogen resistance and mate selection. Research in fish, birds and mammals has suggested that individuals optimise MHC diversity, and therefore offspring fitness, when choosing mates. In reptiles, however, it is unclear whether female mate choice is based on genome-wide genetic characteristics such as microsatellite DNA loci, particular functional-trait loci (e.g., MHC) or both, and MHC's effects on mate choice remain relatively understudied. Herein, we used 13 microsatellite loci and two MHC class I loci to investigate female mate choice of Chinese Alligators (Alligator Sinensis) in the semi-natural condition. We also determined correlations between the MHC genotype of breeding males and male reproductive success. We found that MHC-heterozygous males harbour a greater reproductive success, which probably is the reason that these males are more preferred by the females than MHC-homozygous males. Furthermore, the MHC class I amino-acid distance and functional distance of true mating pairs were higher compared with those of randomly sampled pairs. Analysis of microsatellites revealed that, despite mate choice, females did not completely avoid inbreeding. These findings are the first evidence of MHC-associated mate choice in Chinese Alligators, suggesting that females may adopt different mating strategies after assessing the MHC characteristics of potential mates.
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male density a signal for population self regulation in Alligator Sinensis
Proceedings of The Royal Society B: Biological Sciences, 2019Co-Authors: Lan Zhao, Liming Fang, Qiuhong Wan, Shengguo FangAbstract:The regulation of population density is suggested to be indirect and occurs with a time-lag effect, as well as being female centred. Herein, we present a quantitative analysis on the precise, timel...
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the sex ratio of wild chinese Alligators Alligator Sinensis
Current Zoology, 2013Co-Authors: Lan Zhao, Haiqiong Yang, Liming Fang, Shengguo FangAbstract:The Chinese Alligator Alligator Sinensis is one of the most endangered crocodilian species, and typically exhibits temperature-dependent sex determination. It is extremely important to clarify the sex structure of Chinese Alligators to implement recovery projects successfully. However, the sex ratio of wild Chinese Alligators remains unknown. In this study, we collected 28 years of sex ratio data from Chinese Alligators residing in the natural and artificial habitats of Changxing Nature Reserve, China, and examined the differences in the sex ratio dynamics between these two populations. We observed that the sex ratio of wild Chinese Alligators is 1 male to 4.507 females, which was significantly lower compared to that of the captive population (1 to 2.040; P 0.05). Overall, this study indicates that the stabilized fe- male-biased sex ratio of Changxing Chinese Alligators might result from selection pressure caused by local mate competition and major inbreeding (Current Zoology 59 (6): 725-731, 2013).
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a bacterial artificial chromosome library for the chinese Alligator Alligator Sinensis
Gene, 2012Co-Authors: Ke He, Qing Ye, Hui Chen, Shengguo FangAbstract:Abstract Chinese Alligator (Alligator Sinensis) is a rare and endangered species endemic to China. To better understand genetic details of the Chinese Alligator genomic structure, a highly redundant bacterial artificial chromosome (BAC) library was constructed. This library consists of 216,238 clones with an average insert size of about 90 kb, indicating that the library contains 6.8-fold genome equivalents. Subsequently, we constructed a 516 kb contig map for the Chinese Alligator olfactory receptor (OR) genes, which spans nine BAC clones, and subjected the BACs to full sequencing. The sequence analysis revealed that this contig contained 16 OR functional genes and meanwhile demonstrated that the nine BACs, which constituted the contig, overlapped correctly, proving the usability of this genome library. As a result, this BAC library could provide a useful platform for physical mapping, genome sequencing or complex analysis of targeted genomic regions for this rare species.
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establishment and cryopreservation of liver heart and muscle cell lines derived from the chinese Alligator Alligator Sinensis
Chinese Science Bulletin, 2011Co-Authors: Changjun Zeng, Qing Ye, Shengguo FangAbstract:The Chinese Alligator, Alligator Sinensis, is a critically endangered species. A conservation project of gene resources for an endangered species first involves the preservation of organs, tissues, gametes, genomic DNA libraries and cell lines. The present study is the first to establish and cryopreserve cell lines of liver, heart and muscle tissues from the Chinese Alligator. The study revealed that there was a large discrepancy in cell migration time in primary cultures among liver (11–12 d), heart (13–14 d) and muscle (17–18 d) tissue pieces. The differences in time in primary cell culture suggested that it was relatively easy to build visceral-derived cell lines for reptiles. Biological analysis showed that the population doubling time for thawed cells was approximately 36 h. Karyotyping revealed that the frequency of Chinese Alligator cells showing chromosome number as 2n=32 was 88.6%–93.4%. Chinese Alligator cell lines established here provide a vital resource for research and are likely to be useful for protection of this rare and critically endangered species. Furthermore, the establishment of these methods may supply technical and theoretical support for preserving genetic resources at the cellular level for other reptile species.
Yongkang Zhou - One of the best experts on this subject based on the ideXlab platform.
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the first report of cryptosporidium testudinis in chinese Alligators Alligator Sinensis in china
Parasitology Research, 2020Co-Authors: Jiamin Huang, Yongkang Zhou, Huiliang Chen, Song Wang, Qi Ren, Zhui Fang, Kelei Zheng, Xinchao LiuAbstract:Several Cryptosporidium species that infect reptiles, especially squamates, are well described, but there is limited data about Cryptosporidium species infecting crocodilians. In this study, we assess the occurrence of intestinal parasites using traditional microscopic examination and describe the prevalence and Cryptosporidium species in the captive-bred Chinese Alligators (Alligator Sinensis) in eastern China using molecular methods. The results of microscopic examination showed that no intestinal parasites were detected among the 491 fecal samples examined from the Chinese Alligators. The overall prevalence for Cryptosporidium was 0.41% (2/491) by PCR detection using the SSU rRNA locus. Sequence and phylogenetic analysis of the SSU rRNA, COWP, and actin genes revealed the presence of Cryptosporidium testudinis, which has been isolated primarily from chelonians. This is the first detection of the specific DNA of C. testudinis in the feces of the Chinese Alligator. This study expands our knowledge of the Cryptosporidium species involved in crocodiles, and more extensive studies are necessary to confirm the validity of C. testudinis in crocodiles.
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cloning characterisation and expression profile of kisspeptin1 and the kisspeptin1 receptor in the hypothalamic pituitary ovarian axis of chinese Alligator Alligator Sinensis during the reproductive cycle
Reproduction Fertility and Development, 2020Co-Authors: Yongkang Zhou, Ali Izaz, Shulong Duan, Ruidong Zhang, Renping Wang, Xiaobing WuAbstract:Kisspeptin1 (Kiss1), a product of the Kiss1 gene, plays an important role in the regulation of reproduction in vertebrates by activating the Kiss1 receptor (Kiss1R) and its coexpression with gonadotrophin-releasing hormone (GnRH) in GnRH neurons. The purpose of this study was to clone the Kiss1 and Kiss1R genes found in the brain of Alligator Sinensis and to explore their relationship with reproduction. The full-length cDNA of Kiss1 is 816bp, the open reading frame (ORF) is 417bp and the gene encodes a 138-amino acid precursor protein. The full-length cDNA of Kiss1R is 2348bp, the ORF is 1086bp and the gene encodes a 361-amino acid protein. Quantitative polymerase chain reaction showed that, except for Kiss1R expression in the hypothalamus, the expression of Kiss1 and Kiss1Rduring the reproductive period of A. Sinensis was higher than that in the hypothalamus, pituitary gland and ovary during the hibernation period. The changes in GnRH2 mRNA in the hypothalamus were similar to those of GnRH1 and peaked during the reproductive period. This study confirms the existence of Kiss1 and Kiss1R in A. Sinensis and the findings strongly suggest that Kiss1 and Kiss1R may participate in the regulation of GnRH secretion in the hypothalamus of Alligators during the reproductive period. Furthermore, this is the first report of the full-length cDNA sequences of Kiss1 and Kiss1R in reptiles.
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cytochemical patterns of the peripheral blood cells in chinese Alligator Alligator Sinensis
Tissue & Cell, 2018Co-Authors: Xianxian Chen, Yongkang Zhou, En Li, Fei Peng, Jing Wang, Shengzhou ZhangAbstract:Abstract The Chinese Alligator ( Alligator Sinensis ) is an endemic rare crocodilian species in China. In this study, we investigated the cytochemical patterns of peripheral blood cells in Chinese Alligators for the first time by a range of cytochemical staining techniques including periodic acid-Schiff(PAS), sudan black B(SBB), peroxidase(POX), alkaline phosphatase(AKP), acid phosphatase(ACP), chloroacetic acid AS-D naphthol esterase(AS-D) and α-naphthol acetate esterase(α-NAE) staining. Erythrocytes were positive for PAS, and negative for all the other staining; heterophils were strongly positive for SBB, POX, ACP, AKP, AS-D and α-NAE, while weakly positive for PAS staining; eosinophils were strongly positive for PAS, POX, AKP, ACP and AS-D, and weakly positive for SBB and α-NAE staining; basophils were strongly positive for PAS, positive for POX, ACP, AKP and α-NAE, and weakly positive for AS-D, while negative for SBB staining; monocytes were weakly positive for PAS, ACP, AKP and α-NAE, while negative for SBB, POX and AS-D staining; lymphocytes were weakly positive for PAS and α-NAE, negative for all the other staining; thrombocytes were weakly positive for PAS, and negative for all the other staining. Our results add up knowledge about Chinese Alligator blood cells.
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molecular cloning of androgen receptor and gene expression of sex steroid hormone receptors in the brain of newborn chinese Alligator Alligator Sinensis
Gene, 2018Co-Authors: Yongkang Zhou, Ali Izaz, Mengjuan Wu, Ruidong Zhang, Renping Wang, Ying Zhang, Xiaobing WuAbstract:Abstract Sex steroid hormones play an important role in mediating physiological responses and developmental processes through their receptors across all vertebrates. Chinese Alligator (Alligator Sinensis) is a critically endangered reptile species unique to China. In this study, we have cloned one of the sex steroid hormone receptor genes, androgen receptor (AR) from the brain of Chinese Alligator for the first time. The full-length AR cDNA is 2717 bp in length with an open reading frame (ORF) encoding 722 amino acids. Amino acid alignment analyses indicated that the ARs exhibit highly conserved functional domains. Especially, the P-box and D-box, which are essential to ensure that receptor binding to the androgen response elements, are completely conserved in selected species. Using the quantitative real-time PCR (qPCR), the spatial expression of four receptor mRNAs in all newborn brain tissues and temporal expression of them in the cerebrum during the embryonic development in Chinese Alligators were investigated. The results of qPCR showed ubiquitous expression of the four receptor mRNAs in all newborn brain tissues examined and significant changes in the expression levels of these receptor mRNAs in the embryonic development. These results suggest that sex steroid hormones might play an important role in the regulation of complex neuroendocrine activities in newborn Chinese Alligator. Furthermore, these data provide an important foundation for further studies on endocrinology and molecular biology of non-mammalian sex steroid hormone receptors.
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insights into the seasonal adaptive mechanisms of chinese Alligators Alligator Sinensis from transcriptomic analyses
Australian Journal of Zoology, 2018Co-Authors: Fang Zhang, Yongkang Zhou, En Li, Rong Wu, Xiaobing WuAbstract:The Chinese Alligator (Alligator Sinensis) is an endemic and rare species in China, and is considered to be one of the most endangered vertebrates in the world. It is known to hibernate, an energy-saving strategy against cold temperatures and food deprivation. Changes in gene expression during hibernation remain largely unknown. To understand these complex seasonal adaptive mechanisms, we performed a comprehensive survey of differential gene expression in heart, skeletal muscle, and kidney of hibernating and active Chinese Alligators using RNA-Sequencing. In total, we identified 4780 genes differentially expressed between the active and hibernating periods. GO and KEGG pathway analysis indicated the likely role of these differentially expressed genes (DEGs). The upregulated DEGs in the active Chinese Alligator, CSRP3, MYG and PCKGC, may maintain heart and skeletal muscle contraction, transport and storage of oxygen, and enhance the body’s metabolism, respectively. The upregulated DEGs in the dormant Chinese Alligator, ADIPO, CIRBP and TMM27, may improve insulin sensitivity and glucose/lipid metabolism, protect cells against harmful effects of cold temperature and hypoxia, regulate amino acid transport and uptake, and stimulate the proliferation of islet cells and the secretion of insulin. These results provide a foundation for understanding the molecular mechanisms of the seasonal adaptation required for hibernation in Chinese Alligators, as well as effective information for other non-model organisms research.
Ruidong Zhang - One of the best experts on this subject based on the ideXlab platform.
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cloning characterisation and expression profile of kisspeptin1 and the kisspeptin1 receptor in the hypothalamic pituitary ovarian axis of chinese Alligator Alligator Sinensis during the reproductive cycle
Reproduction Fertility and Development, 2020Co-Authors: Yongkang Zhou, Ali Izaz, Shulong Duan, Ruidong Zhang, Renping Wang, Xiaobing WuAbstract:Kisspeptin1 (Kiss1), a product of the Kiss1 gene, plays an important role in the regulation of reproduction in vertebrates by activating the Kiss1 receptor (Kiss1R) and its coexpression with gonadotrophin-releasing hormone (GnRH) in GnRH neurons. The purpose of this study was to clone the Kiss1 and Kiss1R genes found in the brain of Alligator Sinensis and to explore their relationship with reproduction. The full-length cDNA of Kiss1 is 816bp, the open reading frame (ORF) is 417bp and the gene encodes a 138-amino acid precursor protein. The full-length cDNA of Kiss1R is 2348bp, the ORF is 1086bp and the gene encodes a 361-amino acid protein. Quantitative polymerase chain reaction showed that, except for Kiss1R expression in the hypothalamus, the expression of Kiss1 and Kiss1Rduring the reproductive period of A. Sinensis was higher than that in the hypothalamus, pituitary gland and ovary during the hibernation period. The changes in GnRH2 mRNA in the hypothalamus were similar to those of GnRH1 and peaked during the reproductive period. This study confirms the existence of Kiss1 and Kiss1R in A. Sinensis and the findings strongly suggest that Kiss1 and Kiss1R may participate in the regulation of GnRH secretion in the hypothalamus of Alligators during the reproductive period. Furthermore, this is the first report of the full-length cDNA sequences of Kiss1 and Kiss1R in reptiles.
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molecular cloning of androgen receptor and gene expression of sex steroid hormone receptors in the brain of newborn chinese Alligator Alligator Sinensis
Gene, 2018Co-Authors: Yongkang Zhou, Ali Izaz, Mengjuan Wu, Ruidong Zhang, Renping Wang, Ying Zhang, Xiaobing WuAbstract:Abstract Sex steroid hormones play an important role in mediating physiological responses and developmental processes through their receptors across all vertebrates. Chinese Alligator (Alligator Sinensis) is a critically endangered reptile species unique to China. In this study, we have cloned one of the sex steroid hormone receptor genes, androgen receptor (AR) from the brain of Chinese Alligator for the first time. The full-length AR cDNA is 2717 bp in length with an open reading frame (ORF) encoding 722 amino acids. Amino acid alignment analyses indicated that the ARs exhibit highly conserved functional domains. Especially, the P-box and D-box, which are essential to ensure that receptor binding to the androgen response elements, are completely conserved in selected species. Using the quantitative real-time PCR (qPCR), the spatial expression of four receptor mRNAs in all newborn brain tissues and temporal expression of them in the cerebrum during the embryonic development in Chinese Alligators were investigated. The results of qPCR showed ubiquitous expression of the four receptor mRNAs in all newborn brain tissues examined and significant changes in the expression levels of these receptor mRNAs in the embryonic development. These results suggest that sex steroid hormones might play an important role in the regulation of complex neuroendocrine activities in newborn Chinese Alligator. Furthermore, these data provide an important foundation for further studies on endocrinology and molecular biology of non-mammalian sex steroid hormone receptors.
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molecular cloning of esr2 and gene expression analysis of esr1 and esr2 in the pituitary gland of the chinese Alligator Alligator Sinensis during female reproductive cycle
Gene, 2017Co-Authors: Ruidong Zhang, Yongkang Zhou, Rong Wu, Xiaobing WuAbstract:Abstract Estrogens play critical roles in reproductive physiology via estrogen receptors (ESRs) in vertebrates, including reptiles. Chinese Alligator (Alligator Sinensis) is an endemic and endangered reptile species in China. In the present study, we cloned ESR2 gene from the ovary using rapid amplification of cDNA ends (RACE), investigated the spatial expression of ESRs in various tissues and temporal expression of ESRs in the pituitary glands during the reproductive cycle in Chinese Alligators by quantitative real-time PCR (qPCR). Bioinformatics and phylogenetic analysis of deduced ESR2 protein were also performed. The full-length cDNA of the ESR2 is 1647 bp in length, with an open-reading frame encoding 548 amino acids. The bioinformatics analysis indicated that the deduced amino acid sequence of Alligator ESR2 was highly conserved with that of other vertebrate species. In addition, compared to human ESR2, the 14 amino acids in the Alligator ESR2 that are essential for specific recognition of estradiol are entirely conserved. The phylogenetic analysis showed that Alligators were more closely related to birds than to other reptiles. The results of qPCR showed that the tissue distribution patterns of both ESR subtype mRNAs appeared to be different. In male tissues, the highest mRNA level of both ESRs is in the liver. While in female tissues, ESR1 and ESR2 showed the highest mRNA level in the hypothalamus and pituitary gland, respectively. During the female reproductive cycle, the expression level of ESR1 mRNA increased from the initial post-hibernation period to the reproductive period, reached its peak in the reproductive period, and then decreased in the autumn active period and hibernation period. Conversely, the highest transcription level of ESR2 was observed in the hibernation period.
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immunohistochemical localization of somatostatin in the brain of chinese Alligator Alligator Sinensis
Anatomical Record-advances in Integrative Anatomy and Evolutionary Biology, 2017Co-Authors: Huan Wang, Yongkang Zhou, Shengzhou Zhang, Ruidong Zhang, Xiaobing WuAbstract:In this study, the regional distribution and histological localization of somatostatin (SS) immunoreactive (IR) perikarya and fibers was investigated for the first time in the brain of adult Chinese Alligator by immunohistochemical method. The results showed SS-IR perikarya and fibers were widely distributed in various parts of the brain except for olfactory bulbs. In the telencephalon, SS-IR perikarya were predominantly located in the cellular layer and deep plexiform layer of dorsomedial and medial cortex, less in the dorsal and lateral cortex, while SS-IR fibers were found in all layers of the cerebral cortex. SS-IR perikarya and fibers were also detected in the dorsal ventricular ridge, hippocampus cortex, accessory olfactory bulb nuclearus, lenticular nucleus and caudate nucleus. In the diencephalon, SS-IR perikarya and fibers were mainly present in supraoptic nucleus, paraventricular nucleus of hypothalamus, recessus infundibular nucleus, median eminence, the pineal gland and pituitary gland, in which the IR-fibers were abundant, appearing dot-shaped and varicosity-like. In the mesencephalon, they were present in tectum cortex, ependyma of cerebral aqueduct and the periaqueductal grey matter. Additionally, they were also detected in Purkinje's cellular layer of cerebellum, in the reticularis nucleus and raphe nucleus of medulla oblongata. The distribution pattern of SS-IR perikarya and fibers in the brain of Chinese Alligator is generally similar to that reported in other reptiles, but also has some specific features. The wide distribution indicated that SS might be a neurotransmitter or neuromodulator which acts on many kinds of target cells with a wide range of physiological functions. This article is protected by copyright. All rights reserved.
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molecular cloning characterization tissue distribution and mrna expression changes during the hibernation and reproductive periods of estrogen receptor alpha esr1 in chinese Alligator Alligator Sinensis
Comparative Biochemistry and Physiology B, 2016Co-Authors: Ruidong Zhang, Yongkang Zhou, Yuehong Hu, Huan Wang, Rong Wu, Xiaobing WuAbstract:Abstract Chinese Alligator, Alligator Sinensis , is a critically endangered reptile species unique to China. Little is known about the mechanism of growth- and reproduction-related hormones gene expression in Chinese Alligator. Estrogens play important roles in regulating multiple reproduction- and non-reproduction-related functions by binding to their corresponding receptors. Here, the full-length cDNA of estrogen receptor alpha ( ERα / ESR1 ) was cloned and sequenced from Chinese Alligator for the first time, which comprises 1764 bp nucleotides and encodes a predicted protein of 587 amino acids. Phylogenetic analysis of ESR1 showed that crocodilians and turtles were the sister-group of birds. The results of real-time quantitative PCR indicated that the ESR1 mRNA was widely expressed in the brain and peripheral tissues. In the brain and pituitary gland, ESR1 was most highly transcribed in the cerebellum. But in other peripheral tissues, ESR1 mRNA expression level was the highest in the ovary. Compared with hibernation period, ESR1 mRNA expression levels were increased significantly in the reproductive period ( P P > 0.05). The ESR1 mRNA expression levels changes during the two periods of different tissues suggested that ESR1 might play an important role in mediation of estrogenic multiple reproductive effects in Chinese Alligator. Furthermore, it was the first time to quantify ESR1 mRNA level in the brain of crocodilians, and the distribution and expression of ESR1 mRNA in the midbrain, cerebellum and medulla oblongata was also reported for the first time in reptiles.
Renping Wang - One of the best experts on this subject based on the ideXlab platform.
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cloning characterisation and expression profile of kisspeptin1 and the kisspeptin1 receptor in the hypothalamic pituitary ovarian axis of chinese Alligator Alligator Sinensis during the reproductive cycle
Reproduction Fertility and Development, 2020Co-Authors: Yongkang Zhou, Ali Izaz, Shulong Duan, Ruidong Zhang, Renping Wang, Xiaobing WuAbstract:Kisspeptin1 (Kiss1), a product of the Kiss1 gene, plays an important role in the regulation of reproduction in vertebrates by activating the Kiss1 receptor (Kiss1R) and its coexpression with gonadotrophin-releasing hormone (GnRH) in GnRH neurons. The purpose of this study was to clone the Kiss1 and Kiss1R genes found in the brain of Alligator Sinensis and to explore their relationship with reproduction. The full-length cDNA of Kiss1 is 816bp, the open reading frame (ORF) is 417bp and the gene encodes a 138-amino acid precursor protein. The full-length cDNA of Kiss1R is 2348bp, the ORF is 1086bp and the gene encodes a 361-amino acid protein. Quantitative polymerase chain reaction showed that, except for Kiss1R expression in the hypothalamus, the expression of Kiss1 and Kiss1Rduring the reproductive period of A. Sinensis was higher than that in the hypothalamus, pituitary gland and ovary during the hibernation period. The changes in GnRH2 mRNA in the hypothalamus were similar to those of GnRH1 and peaked during the reproductive period. This study confirms the existence of Kiss1 and Kiss1R in A. Sinensis and the findings strongly suggest that Kiss1 and Kiss1R may participate in the regulation of GnRH secretion in the hypothalamus of Alligators during the reproductive period. Furthermore, this is the first report of the full-length cDNA sequences of Kiss1 and Kiss1R in reptiles.
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molecular cloning of androgen receptor and gene expression of sex steroid hormone receptors in the brain of newborn chinese Alligator Alligator Sinensis
Gene, 2018Co-Authors: Yongkang Zhou, Ali Izaz, Mengjuan Wu, Ruidong Zhang, Renping Wang, Ying Zhang, Xiaobing WuAbstract:Abstract Sex steroid hormones play an important role in mediating physiological responses and developmental processes through their receptors across all vertebrates. Chinese Alligator (Alligator Sinensis) is a critically endangered reptile species unique to China. In this study, we have cloned one of the sex steroid hormone receptor genes, androgen receptor (AR) from the brain of Chinese Alligator for the first time. The full-length AR cDNA is 2717 bp in length with an open reading frame (ORF) encoding 722 amino acids. Amino acid alignment analyses indicated that the ARs exhibit highly conserved functional domains. Especially, the P-box and D-box, which are essential to ensure that receptor binding to the androgen response elements, are completely conserved in selected species. Using the quantitative real-time PCR (qPCR), the spatial expression of four receptor mRNAs in all newborn brain tissues and temporal expression of them in the cerebrum during the embryonic development in Chinese Alligators were investigated. The results of qPCR showed ubiquitous expression of the four receptor mRNAs in all newborn brain tissues examined and significant changes in the expression levels of these receptor mRNAs in the embryonic development. These results suggest that sex steroid hormones might play an important role in the regulation of complex neuroendocrine activities in newborn Chinese Alligator. Furthermore, these data provide an important foundation for further studies on endocrinology and molecular biology of non-mammalian sex steroid hormone receptors.
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analysis of tcrβ and tcrγ genes in chinese Alligator provides insights into the evolution of tcr genes in jawed vertebrates
Developmental and Comparative Immunology, 2018Co-Authors: Xifeng Wang, Chaolin Wang, Kongpan Li, Cuncun Ke, Yaofeng Zhao, Di Yu, Renping Wang, Peng Wang, Xin ZhouAbstract:Abstract All jawed vertebrates have four T cell receptor (TCR) chains that are expressed by thymus-derived lymphocytes and play a major role in animal immune defence. However, few studies have investigated the TCR chains of crocodilians compared with those of birds and mammals, despite their key evolutionary position linking amphibians, reptiles, birds and mammals. Here, employing an Alligator Sinensis genomic bacterial artificial chromosome (BAC) library and available genome data, we characterized the genomic organization, evolution and expression of TRB and TRG loci in Alligator Sinensis . According to the sequencing data, the Alligator Sinensis TRB locus spans approximately 500 Kb of genomic DNA containing two D-J-C clusters and 43 V gene segments and is organized as Vβ (39) -pJβ1-pCβ1-pDβ1-Dβ2- Jβ2 (12) -Cβ2-Vβ (4) , whereas the TRG locus spans 115 Kb of DNA genomic sequence consisting of 18 V gene segments, nine J gene segments and one C gene segment and is organized in a classical translocon pattern as Vγ (18) –Jγ (9) -Cγ. Moreover, syntenic analysis of TRB and TRG chain loci suggested a high degree of conserved synteny in the genomic regions across mammals, birds and Alligator Sinensis. By analysing the cloned TRB / TRG cDNA, we identified the usage pattern of V families in the expressed TRB and TRG . An analysis of the junctions of the recombined VJ revealed the presence of N and P nucleotides in both expressed TRB and TRG sequences. Phylogenetic analysis revealed that TRB and TRG loci possess distinct evolutionary patterns. Most Alligator Sinensis V subgroups have closely related orthologues in chicken and duck, and a small number of Alligator Sinensis V subgroups have orthologues in mammals, which supports the hypothesis that crocodiles are the closest relatives of birds and mammals. Collectively, these data provide insights into TCR gene evolution in vertebrates and improve our understanding of the Alligator Sinensis immune system.
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as cath1 6 novel cathelicidins with potent antimicrobial and immunomodulatory properties from Alligator Sinensis play pivotal roles in host antimicrobial immune responses
Biochemical Journal, 2017Co-Authors: Yan Chen, Renping Wang, Shasha Cai, Xue Qiao, Zhilai Guo, Yiqun Kuang, Yipeng WangAbstract:Crocodilians are regarded as possessing a powerful immune system. However, the composition and action of the crocodilian immune system have remained unclear until now. Cathelicidins, the principal family of host defense peptides, play pivotal roles in vertebrate immune defense against microbial invasions. However, cathelicidins from crocodilians have not been extensively studied to date. In the present study, six novel cathelicidins (As-CATH1-6) were identified and characterized from the endangered Chinese Alligator (Alligator Sinensis). As-CATH1-6 exhibit no sequence similarity with any of the known cathelicidins. Structure analysis indicated that As-CATH1-3 adopt a random coil secondary conformation, whereas As-CATH4-6 were predicted to mainly adopt an amphipathic α-helix conformation. Among them, As-CATH4-6 exhibited potent, broad-spectrum and rapid antimicrobial activity by inducing the disruption of cell membrane integrity. They also exhibited strong ability to prevent the formation of bacterial biofilms and eradicate preformed biofilms. Furthermore, As-CATH4-6 exhibited potent anti-inflammatory activity by inhibiting the lipopolysaccharide (LPS)-induced production of nitric oxide (NO) and pro-inflammatory cytokines in mouse peritoneal macrophages. They directly neutralized LPS toxicity and therefore inhibited the binding of LPS to the TLR4 receptor and the subsequent activation of inflammatory response pathways. In a peritonitis mice model, As-CATH2-6 provided effective protection against bacterial infection through enhanced immune cell recruitment. In the host Chinese Alligator, As-CATH1-6 are mainly expressed in immune organs and epithelial tissues. Bacterial infection significantly enhances their expression, which implies an important role in host anti-infective response. Taken together, the diversity and multiple functions of As-CATH1-6 partially reveal the powerful immune system of the Chinese Alligator.
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temporal profile of nerve growth factor expression in the partial central nervous system of the yangtze Alligator Alligator Sinensis reptilia crocodylia during early postnatal growth
Anatomical Record-advances in Integrative Anatomy and Evolutionary Biology, 2013Co-Authors: Lanrong Zheng, Renping Wang, Fangfang Chen, Yongkang ZhouAbstract:Expression of nerve growth factor (NGF) in structures of the partial central nervous system of the Yangtze Alligator, Alligator Sinensis (Reptilia, Crocodylia) was examined during early postnatal growth using immunohistochemistry and Western blot assays. In animals 0–2 years of age NGF-positive cells in the cerebral cortex increased gradually in number and size, and were predominantly distributed in the molecular layer. NGF-positive cells in the midbrain showed similar increases but with predominant distribution in the ependymal layer. NGF-positive cells increased in the cerebellum between 0 and 1 years of age, with increased NGF expression being seen during the first 2 years of life mostly in the ependymal layer. NGF-positive cells were mainly found in the gray matter of the spinal cord with decreasing cell numbers, NGF expression levels being seen from 0 to 2 years and small processes without synaptic connection from 1 to 2 years. These results suggest that NGF is involved in the early postnatal growth of several structures of Yangtze Alligator partial central nervous system, suggesting a possible role of NGF in the Yangtze Alligator partial central nervous system. Anat Rec, 296:840–845, 2013. © 2013 Wiley Periodicals, Inc.