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Morphological and Hormonal Identifi cation of Porcine Atretic Follicles and Relationship Analysis of Hormone Receptor Levels During Granulosa Cell Apoptosis In vivo

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Journal of Integrative Agriculture 2014年第5期13卷 1058-1064页

作者： YU De-bing YU Min-li LIN Fei JIANG Bao-chun YANG Li-na WANG Si-yu ZHAO Ying WNAG Zheng-chao Department of animal Genetics Breeding and Reproduction College of Animal Science and Technology Nanjing Agricultural University

Recent reports have demonstrated that follicular atresia is initiated or caused by granulosa cell apoptosis followed by theca cell degeneration in mammalian ovaries, but the mechanism of follicular atresia is still to be elucidated. Therefore, our present study was designed to examine our hypothesis that the changes of follicular microenvironment induce the granulosa cell apoptosis during pocrine follicular atresia in vivo. We firstly isolated intact porcine antral follicles and identified them into three groups, healthy follicles （HF）, early atretic follicles （EAF） and progressed atretic follicles （PAF） through morphology and histology. To further confirm their status, we detected hormone levels in follicular fluids and the expression level of apoptosis gene Bax in granulosa cells. The rate of progesterone （P） and estradiol （E2） was increased with the expression of Bax, indicating hormone can be used as a marker of granulosa cell apoptosis or follicular atresia. Finally, we analyzed the expression level of hormone receptor genes in granulosa cells and their relationship with follicular atresia. In PAF, the expression of Progesterone receptor （PGR） was increased significantly while estradiol receptor （ER） had no notable changes, which suggesting the increased-PGR accelerated the effect of P-stimulated granulosa cell apoptosis. The dramatic increasing of androgen receptor （AR） expression in PAF and the obvious increase of tumor necrosis factor-u receptor （TNFR） in EAF indicated that there are different pathways regulating granulosa cell apoptosis during follicular atresia. Together, our results suggested that different pathways of granulosa cell apoptosis was induced by changing the follicular microenvironment during follicular atresia.

关键词： granulosa cell apoptosis follicular atresia progesterone estradiol porcine

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m^(6)A demethylase FTO regulate CTNNB1 to promote adipogenesis of chicken preadipocyte

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Journal of animal Science and Biotechnology 2023年第2期14卷 578-592页

作者： Kan Li Weichen Huang Zhijun Wang Qinghua Nie Department of animal Genetics Breeding and ReproductionCollege of Animal ScienceSouth China Agricultural UniversityGuangzhou 510642China National‑Local Joint Engineering research Center for Livestock breeding Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breedingand Key Laboratory of Chicken GeneticsBreeding and ReproductionMinistry of AgricultureGuangzhou 510642China

Background:N6-methyladenosine(m^(6)A)is an abundant post-transcriptional RNA modification that affects various biological *** fat mass and obesity-associated(FTO)protein,a demethylase encoded by the FTO gene,has been found to regulate adipocyte development in an m^(6)A-dependent manner in multiple ***,the effects of the m^(6)A methylation and FTO demethylation functions on chicken adipogenesis remain *** study aims to explore the association between m^(6)A modification and chicken adipogenesis and the underlying mechanism by which FTO affects chicken preadipocyte ***:The association between m^(6)A modification and chicken lipogenesis was assessed by treating chicken pread-ipocytes with different doses of methyl donor betaine and methylation inhibitor *** results showed that betaine significantly increased methylation levels and inhibited lipogenesis,and the inverse effect was found in preadipocytes after cycloleucine *** of FTO significantly inhibited m^(6)A levels and promoted proliferation and differentiation of chicken *** FTO showed opposite ***,FTO overexpression increased the expression of catenin beta 1(CTNNB1)by improving RNA stability in an m^(6)A-dependent manner,and we proved that FTO could directly target ***,CTNNB1 may be a positive regulator of adipogenesis in chicken ***:m^(6)A methylation of RNA was negatively associated with adipogenesis of chicken *** could regulate CTNNB1 expression in a demethylation manner to promote lipogenesis.

关键词： Adipogenesis Chicken CTNNB1 FTO m^(6)A

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The proteome of IVF-induced aberrant embryo-maternal crosstalk by implantation stage in ewes

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Journal of animal Science and Biotechnology 2020年第2期11卷 393-409页

作者： Qianying Yang Wei Fu Yue Wang Kai Miao Haichao Zhao Rui Wang Min Guo Zhilong Wang Jianhui Tian Lei An Key Laboratory of animal Genetics Breeding and Reproduction of the Ministry of AgricultureNational Engineering Laboratory for Animal BreedingCollege of Animal Science and TechnologyChina Agricultural UniversityBeijing 100193China

Background:Implantation failure limits the success of in vitro fertilization and embryo transfer(IVF-ET).Wellorganized embryo-maternal crosstalk is essential for successful *** studies mainly focused on the aberrant development of in vitro fertilized(IVF)*** contrast,the mechanism of IVF-induced aberrant embryo-maternal crosstalk is not well ***:In the present study,using ewes as the model,we profiled the proteome that features aberrant IVF embryo-maternal crosstalk following *** comparing in vivo(IVO)and IVF conceptuses,as well as matched endometrial caruncular(C)and intercaruncular(IC)areas,we filtered out 207,295,and 403 differentially expressed proteins(DEPs)in each *** functional analysis showed that the IVF conceptuses were characterized by the increased abundance of energy metabolism and proliferation-related proteins,and the decreased abundance of methyl metabolism-related *** addition,IVF endometrial C areas showed the decreased abundance of endometrial remodeling and redox homeostasis-related proteins;while IC areas displayed the aberrant abundance of protein homeostasis and extracellular matrix(ECM)interaction-related *** on these observations,we propose a model depicting the disrupted embryo-maternal crosstalk following IVF-ET:Aberrant energy metabolism and redox homeostasis of IVF embryos,might lead to an aberrant endometrial response to conceptus-derived pregnancy signals,thus impairing maternal *** turn,the suboptimal uterine environment might stimulate a compensation effect of the IVF conceptuses,which was revealed as enhanced energy metabolism and ***:Systematic proteomic profiling provides insights to understand the mechanisms that underlie the aberrant IVF embryo-maternal *** might be helpful to develop practical strategies to prevent implantation failure following IVF-ET.

关键词： Conceptus Embryo-maternal crosstalk Endometrial C areas Endometrial IC areas Ewes HPLC-MS/MS IVF Proteome

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Hepatic steatosis is associated with dysregulated cholesterol metabolism and altered protein acetylation dynamics in chickens

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Journal of animal Science and Biotechnology 2024年第1期15卷 53-67页

作者： Xiaoli Guo Qianqian Zhou Jiaming Jin Fangren Lan Chaoliang Wen Junying Li Ning Yang Congjiao Sun National Engineering Laboratory for animal breeding and Key Laboratory of animal Genetics Breeding and ReproductionMinistry of Agriculture and Rural AffairsChina Agricultural UniversityBeijing 100193China

Background Hepatic steatosis is a prevalent manifestation of fatty liver, that has detrimental effect on the health and productivity of laying hens, resulting in economic losses to the poultry industry. Here, we aimed to systematically investigate the genetic regulatory mechanisms of hepatic steatosis in laying *** Ninety individuals with the most prominent characteristics were selected from 686 laying hens according to the accumulation of lipid droplets in the liver, and were graded into three groups, including the control, mild hepatic steatosis and severe hepatic steatosis groups. A combination of transcriptome, proteome, acetylome and lipidome analyses, along with bioinformatics analysis were used to screen the key biological processes, modifications and lipids associated with hepatic *** The rationality of the hepatic steatosis grouping was verified through liver biochemical assays and RNA-seq. Hepatic steatosis was characterized by increased lipid deposition and multiple metabolic abnormalities. Integration of proteome and acetylome revealed that differentially expressed proteins(DEPs) interacted with differentially acetylated proteins(DAPs) and were involved in maintaining the metabolic balance in the liver. Acetylation alterations mainly occurred in the progression from mild to severe hepatic steatosis, i.e., the enzymes in the fatty acid oxidation and bile acid synthesis pathways were significantly less acetylated in severe hepatic steatosis group than that in mild group(P < 0.05). Lipidomics detected a variety of sphingolipids(SPs) and glycerophospholipids(GPs) were negatively correlated with hepatic steatosis(r ≤-0.5, P < 0.05). Furthermore, the severity of hepatic steatosis was associated with a decrease in cholesterol and bile acid synthesis and an increase in exogenous cholesterol *** In addition to acquiring a global and thorough picture of hepatic steatosis in laying hens, we were able to reveal the role of ac

关键词： Acetylation Cholesterol metabolism Hepatic steatosis Laying hens Multiomics

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Genetic basis of negative heterosis for growth traits in chickens revealed by genome-wide gene expression pattern analysis

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Journal of animal Science and Biotechnology 2021年第4期12卷 1267-1280页

作者： Chunning Mai Chaoliang Wen Zhiyuan Xu Guiyun Xu Sirui Chen Jiangxia Zheng Congjiao Sun Ning Yang National Engineering Laboratory for animal breeding and Key Laboratory of animal Genetics Breeding and ReproductionMinistry of Agriculture and Rural AffairsChina Agricultural UniversityBeijing 100193China

Background:Heterosis is an important biological phenomenon that has been extensively utilized in agricultural ***,negative heterosis is also pervasively observed in nature,which can cause unfavorable impacts on production *** with systematic studies of positive heterosis,the phenomenon of negative heterosis has been largely ignored in genetic studies and breeding programs,and the genetic mechanism of this phenomenon has not been thoroughly elucidated to ***,we used chickens,the most common agricultural animals worldwide,to determine the genetic and molecular mechanisms of negative ***:We performed reciprocal crossing experiments with two distinct chicken lines and found that the body weight presented widely negative heterosis in the early growth of *** heterosis of carcass traits was more common than positive heterosis,especially breast muscle mass,which was over−40%in reciprocal ***-wide gene expression pattern analyses of breast muscle tissues revealed that nonadditivity,including dominance and overdominace,was the major gene inheritance *** genes,including a substantial number of genes encoding ATPase and NADH dehydrogenase,accounted for more than 68%of differentially expressed genes in reciprocal crosses(4257 of 5587 and 3617 of 5243,respectively).Moreover,nonadditive genes were significantly associated with the biological process of oxidative phosphorylation,which is the major metabolic pathway for energy release and animal growth and *** detection of ATP content and ATPase activity for purebred and crossbred progenies further confirmed that chickens with lower muscle yield had lower ATP concentrations but higher hydrolysis activity,which supported the important role of oxidative phosphorylation in negative heterosis for growth traits in ***:These findings revealed that nonadditive genes and their related oxidative phosphorylation were the m

关键词： Chicken Growth Gene expression patterns Heterosis Oxidative phosphorylation

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Identification of single nucleotide polymorphisms of PIK3R1 and DUSP1 genes and their genetic associations with milk production traits in dairy cows

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Journal of animal Science and Biotechnology 2020年第1期11卷 9-19页

作者： Bo Han Yuwei Yuan Lijun Shi Yanhua Li Lin Liu Dongxiao Sun Department of animal Genetics Breeding and ReproductionCollege of Animal Science and TechnologyKey Laboratory of Animal GeneticsBreeding and Reproduction of Ministry of Agriculture and Rural AffairsNational Engineering Laboratory for Animal BreedingChina Agricultural UniversityNo.2 Yuanmingyuan West RoadHaidian DistrictBeijing 100193China Beijing Dairy Cattle Center Beijing 100192China.

Background:Previously,phosphoinositide-3-kinase regulatory subunit 1(PIK3R1)and dual specificity phosphatase 1(DUSP1)were identified as promising candidate genes for milk production traits due to their being differentially expressed between the dry period and the peak of lactation in livers of dairy ***,in this study,the single nucleotide polymorphisms(SNPs)of PIK3R1 and DUSP1 genes were identified and their genetic associations with milk yield,fat yield,fat percentage,protein yield,and protein percentage,were investigated using 1067 Chinese Holstein cows from 40 sire ***:By re-sequencing the entire coding region and 2000 bp of the 5′and 3′flanking regions of the two genes,one SNP in the 5′untranslated region(UTR),three in the 3′UTR,and two in the 3′flanking region of PIK3R1 were identified,and one in the 5′flanking region,one in the 3′UTR,and two in the 3′flanking region of DUSP1 were *** single-locus association analyses showed that five SNPs in PIK3 R1,rs42590258,rs210389799,rs208819656,rs41255622,rs133655926,and rs211408208,and four SNPs in DUSP1,rs207593520,rs208460068,rs209154772,and rs210000760,were significantly associated with milk,fat and protein yields in the first or second lactation(P values≤0.0001 and 0.0461).In addition,by the Haploview 4.2 software,the six and four SNPs in PIK3R1 and DUSP1 respectively formed one haplotype block,and the haplotype-based association analyses showed significant associations between their haplotype combinations and the milk traits in both two lactations(P values≤0.0001 and 0.0364).One SNP,rs207593520(T/G),was predicted to alter the transcription factor binding sites(TFBSs)in the 5′flanking region of ***,the dual-luciferase assay showed that the transcription activity of allele T in rs207593520 was significantly higher than that of allele G,suggesting the activation of transcriptional activity of DUSP1 gene by allele T of ***,the rs207593520 SNP was highlighted as a

关键词： Chinese Holstein DUSP1 Genetic association Milk production PIK3R1 SNP

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Polymorphism of mitochondrial tRNA genes associated with the number of pigs born alive

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Journal of animal Science and Biotechnology 2019年第1期10卷 33-42页

作者： Dan Wang Chao Ning Hai Xiang Xianrui Zheng Minghua Kong Tao Yin Jianfeng Liu Xingbo Zhao National Engineering Laboratory for animal breeding Ministry of Agricultural Key Laboratory of animal Genetics Breeding and Reproduction College of animal Science and Technology China Agricultural University School of Life Science and Engineering Foshan University

Background: Mutations in mitochondrial tRNA genes have been widely reported association with human *** is also important to explore the effect on the number of piglets born alive(NBA).Here,1017 sows were used to investigate the association between polymorphisms in mitochondrial tRNA genes and ***: In total,16 mutations were found in mitochondrial tRNA genes,of which 13 mutations were significantly associated with NBA(P reproductions of mutant carriers were significantly greater than that of wild carriers by 0.989 piglets born alive/sow *** test whether the mutations altered the structure of mitochondrial tRNAs,the secondary and tertiary structures were *** result,C2255 T changed the secondary structure of t RNA-Val by elongating the T stem and shrinking the T loop,and C2255 T and G2259 A in the tRNA-Val gene,C6217 T and T6219 C in the tRNA-Ala gene,and T15283 C in the tRNA-Glu gene altered the tertiary structure of their tRNAs,respectively by changing the folding form of the T arm,and C16487 T in the tRNA-Thr gene changed the tertiary structure of mitochondrial tRNA-Thr by influencing the folding form of the acceptor ***: Results highlight the effect of mitochondrial tRNA genes on the number of piglets born alive,and suggest that polymorphic sites of the tRNA genes be genetic markers for selection of pig reproduction.

关键词： Mitochondrial Number born alive Pig Polymorphism Transfer RNA

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Molecular cloning and characterization of the endothelin 3 gene in black bone sheep

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Journal of animal Science and Biotechnology 2018年第4期9卷 847-857页

作者： Hesham Y.A.Darwish Yuanyuan Zhang Kai Cui Zu Yang Deping Han Xianggui Dong Huaming Mao Weidong Deng Xuemei Deng National Engineering Laboratory for animal breeding and Key Laboratory of animal Genetics Breeding and Reproduction of the Ministry of AgricultureChina Agricultural University animal Production research Institute Agricultural Research Center Ministry of Agriculture and Land Reclamation College of Veterinary Medicine China Agricultural University College of animal Science and Technology Yunnan Agricultural University

Background: Black bone sheep was first discovered in Yunnan province of China in 1970, with unique black pigmentation on the body and internal organs. Endothelin 3(EDN3) has been known as a key gene causing hyperpigmentation in black bone chicken, the Silky ***: In this study, EDN3 was employed as a candidate gene for regulating black color pigmentation. First,EDN3 was cloned from sheep to obtain the full-length cDNA by using the rapid amplification of cDNA ends(RACE).Genomic EDN3 was screened and a total of thirty predicted single nucleotide polymorphisms(SNPs) were genotyped for allele and genotype frequency analysis in a case-control study involving two black bone sheep populations. Genomic copy number analysis of EDN3 in sheep was conducted to measure the variation in copy number. EDN3 expression levels were observed among the groups in adult liver, lymph node, and kidney tissues, as well as embryo kidney samples. Also, among the tissues of black bone and non-black bone ***: The size of the full-length cDNA was 1,578 bp, which included 426 bp of 5′-untranslated region(5′-UTR),an open reading frame(ORF) of 639 bp encoding a protein of 212 amino acids, and a 3′-UTR of 513 bp. Genotype and allele frequencies of all the discovered SNPs were found insignificantly different in black bone and non-black bone sheep(P > 0.05). Genomic copy number analysis of EDN3 in sheep revealed no significant difference between the two sheep groups. No significant variations were found in the adult liver and kidney embryo samples. However,the expression in lymph node and kidney tissue was significantly higher in black bone sheep than that in non-black bone sheep(P < 0.05). Significant variations in the EDN3 expression levels were observed among the tissues of nonblack bone ***: The findings of the present study indicate that unlike in Silky chickens, EDN3 is not responsible for hyperpigmentation but may play a key functional role in immune and excretory

关键词： Black pigmentation Expression level Sequence analysis SNP

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A novel method for rapid and reliable detection of complex vertebral malformation and bovine leukocyte adhesion deficiency in Holstein cattle

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Journal of animal Science and Biotechnology 2012年第3期3卷 130-135页

作者： Yi Zhang Xuehua Fan Dongxiao Sun Yachun Wang Ying Yu Yan Xie Shengli Zhang Yuan Zhang National Engineering Laboratory for animal breeding Key Laboratory of Animal Genetics and Breeding and Reproduction of Ministry of AgricultureCollege of Animal Science and TechnologyChina Agricultural UniversityBeijing 100193China

Background： Complex vertebral malformation （CVM） and bovine leukocyte adhesion deficiency （BLAD） are two autosomal recessive lethal genetic defects frequently occurring in Holstein cattle, identifiable by single nucleotide polymorphisms. The objective of this study is to develop a rapid and reliable genotyping assay to screen the active Holstein sires and determine the carrier frequency of CVM and BLAD in Chinese dairy cattle population. Results： We developed real-time PCR-based assays for discrimination of wild-type and defective alleles, so that carriers can be detected. Only one step was required after the DNA extraction from the sample and time consumption was about 2 hours. A total of 587 Chinese Holstein bulls were assayed, and fifty-six CVM-carriers and eight BLAD-carriers were identified, corresponding to heterozygote carrier frequencies of 9.54% and 1.36%, respectively. The pedigree analysis showed that most of the carriers could be traced back to the common ancestry, Osborndale Ivanhoe for BLAD and Pennstate Ivanhoe Star for CVM. Conclusions： These results demonstrate that real-time PCR is a simple, rapid and reliable assay for BLAD and CVM defective allele detection. The high frequency of the CVM allele suggests that implementing a routine testing system is necessary to gradually eradicate the deleterious gene from the Chinese Holstein population.

关键词： Holstein Cattle BTA1 Leukocyte Adhesion Deficiency Defective Allele Lethal Recessive Allele

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Feed conversion ratio,residual feed intake and cholecystokinin type A receptor gene polymorphisms are associated with feed intake and average daily gain in a Chinese local chicken population

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Journal of animal Science and Biotechnology 2018年第4期9卷 794-802页

作者： Zhenhua Yi Xing Li Wen Luo Zhenqiang Xu Congliang Ji Yan Zhang Qinghua Nie Dexiang Zhang Xiquan Zhang Department of animal Genetics Breeding and ReproductionCollege of Animal ScienceSouth China Agricultural UniversityGuangzhou 510642GuangdongChina Guangdong Provincial Key Lab of Agro-animal Genomics and Molecular breeding and Key Laboratory of Chicken Genetics Breeding and ReproductionMinistry of AgricultureGuangzhou 510642GuangdongChina Wen’s Nanfang Poultry breeding Co.Ltd Yunfu 527400GuangdongChina

Background: The feed conversion ratio(FCR) and residual feed intake(RFI) are common indexes in measuring feed efficiency for livestock. RFI is a feed intake adjusted for requirements for maintenance and production so these two traits are related. Similarly, FCR is related to feed intake and weight gain because it is their ratio. Cholecystokinin type A receptor(CCKAR) plays an important role in animal digestive process. We examined the interplay of these three parameters in a local Chinese chicken ***: The feed intake(FI) and body weights(BW) of 1,841 individuals were monitored on a daily basis from 56 to 105 d of age. There was a strong correlation between RFI and average daily feed intake(ADFI) and a negative correlation between the FCR and daily gain(r=-0.710). Furthermore, we identified 51 single nucleotide polymorphisms(SNPs) in the CCKAR and 4 of these resulted in amino acid mutations. The C334A mutation was specifically associated with FI and the expected feed intake(EFI)(P breeding focus is feed intake. And C334A and G1290A of the CCKAR gene can be deemed as candidate markers for feed intake and weight gain.

关键词： CCKAR Chicken FCR RFI SNP

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