作者： Yang, Anqi;Yan, Saina;Yin, Yanfei;Chen, Chujie;Tang, Xiangwei;...

期刊： International Journal of Molecular Sciences,2023年24(7):6179- ISSN：1661-6596

通讯作者： Maoliang Ran<&wdkj&>Bin Chen

作者机构： [Yan, Saina; Chen, Chujie; Tang, Xiangwei; Yin, Yanfei; Ran, Maoliang; Yang, Anqi; Chen, Bin] Hunan Agr Univ, Coll Anim Sci & Technol, Hunan Prov Key Lab Genet Improvement Domest Anim, Changsha 410128, Peoples R China.

通讯机构： [Maoliang Ran; Bin Chen] C;College of Animal Science and Technology, Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Changsha 410128, China<&wdkj&>Authors to whom correspondence should be addressed.

关键词： adenosine triphosphate;calcium ion;messenger RNA;Myc protein;platelet endothelial cell adhesion molecule 1;prostaglandin E2;small interfering RNA;vasculotropin A;zebularine;guanine nucleotide exchange factor;hydrogen sulfite;sulfite;animal cell;animal tissue;apoptosis;Article;cell proliferation;controlled study;developmental stage;DNA demethylation;DNA methylation;down regulation;exon;fzd7 gene;G2 phase cell cycle checkpoint;gene;gene expression;gene expression regulation;gene silencing;genetic transfection;male;mRNA expression level;newborn;nonhuman;pig;protein expression;regulatory mechanism;RNA interference;RNA sequencing;Sertoli cell;somatic cell;testis development;testis tissue;whole genome bisulfite sequencing;animal;cell proliferation;CpG island;genetics;mammal;Animals;Cell Proliferation;CpG Islands;DNA Methylation;Guanine Nucleotide Exchange Factors;Male;Mammals;Sulfites;Swine

摘要： The regulatory role of non-CpG methylation in mammals has been important in whole-genome bisulfite sequencing. It has also been suggested that non-CpG methylation regulates gene expression to affect the development and health of mammals. However, the dynamic regulatory mechanisms of genome-wide, non-CpG methylation during testicular development still require intensive study. In this study, we analyzed the dataset from the whole-genome bisulfite sequencing (WGBS) and the RNA-seq of precocious porcine testicular tissues across two developmental stages (1 and 75 days old) in order to explore the regulatory roles of non-CpG methylation. Our results showed that genes regulated by non-CpG methylation affect the development of testes in multiple pathways. Furthermore, several hub genes that are regulated by non-CpG methylation during testicular development—such as VEGFA, PECAM1, and FZD7—were also identified. We also found that the relative expression of FZD7 was downregulated by the zebularine-induced demethylation of the first exon of FZD7. This regulatory relationship was consistent with the results of the WGBS and RNA-seq analysis. The immature porcine Sertoli cells were transfected with RNAi to mimic the expression patterns of FZD7 during testicular development. The results of the simulation test showed that cell proliferation was significantly impeded and that cell cycle arrest at the G2 phase was caused by the siRNA-induced FZD7 inhibition. We also found that the percentage of early apoptotic Sertoli cells was decreased by transfecting them with the RNAi for FZD7. This indicates that FZD7 is an important factor in linking the proliferation and apoptosis of Sertoli cells. We further demonstrated that Sertoli cells that were treated with the medium collected from apoptotic cells could stimulate proliferation. These findings will contribute to the exploration of the regulatory mechanisms of non-CpG methylation in testicular development and of the relationship between the proliferation and apoptosis of normal somatic cells. © 2023 by the authors.

语种： 英文

作者： Chen, Chujie;Tang, Xiangwei;Yan, Saina;Yang, Anqi;Xiang, Jiaojiao;...

期刊： International Journal of Molecular Sciences,2023年24(19):14475 ISSN：1661-6596

通讯作者： Chen, Bin;Gu, JJ

作者机构： [Chen, Chujie; Tang, Xiangwei; Yan, Saina; Gu, JJ; Deng, Yanhong; Yang, Anqi; Chen, Bin; Yin, Yulong; Gu, Jingjing; Xiang, Jiaojiao] Hunan Agr Univ, Coll Anim Sci & Technol, Changsha 410128, Peoples R China.;[Chen, Chujie; Tang, Xiangwei; Yan, Saina; Gu, JJ; Deng, Yanhong; Yang, Anqi; Chen, Bin; Gu, Jingjing; Xiang, Jiaojiao] Hunan Agr Univ, Hunan Prov Key Lab Genet Improvement Domest Anim, Changsha 410128, Peoples R China.;[Yin, Yulong] Chinese Acad Sci, Key Lab Agroecol Proc Subtrop Reg, Inst Subtrop Agr, Changsha 410125, Peoples R China.

通讯机构： [Gu, JJ ; Chen, B] H;Hunan Agr Univ, Coll Anim Sci & Technol, Changsha 410128, Peoples R China.;Hunan Agr Univ, Hunan Prov Key Lab Genet Improvement Domest Anim, Changsha 410128, Peoples R China.

关键词： 6 n methyladenosine;activin receptor 2;alkylated DNA repair protein alkB homolog 5;aurora C kinase;cyclic AMP;G protein coupled receptor;hydroxymethylglutaryl coenzyme A reductase kinase;mammalian target of rapamycin;messenger RNA;phosphatidylinositol 3 kinase;protein kinase B;transcription factor Sox9;transcriptome;Wnt protein;phosphatidylinositol 3 kinase;RNA;transcriptome;animal cell;animal experiment;animal tissue;apoptosis;Article;cell proliferation;cell viability;chromosome 1;comparative study;consensus sequence;controlled study;epigenetic modification;gene expression;genetic transcription;high throughput sequencing;KEGG;male;maturity;methylome;nonhuman;oxidation;pig;protein expression;puberty;real time polymerase chain reaction;regulatory mechanism;reproduction;RNA immunoprecipitation;RNA interference;RNA methylation;RNA sequencing;Sertoli cell;sexual maturation;sexual maturity;signal transduction;spermatogenesis;spliceosome;start codon;stop codon;testicular cell line;testis development;testis tissue;animal;epigenome;metabolism;sexual maturation;testis;Animals;Epigenome;Male;Phosphatidylinositol 3-Kinases;RNA;Sexual Maturation;Swine;Testis;Transcriptome

摘要： RNA N6-methyladenosine (m6A) modification is one of the principal post-transcriptional modifications and plays a dynamic role in testicular development and spermatogenesis. However, the role of m6A in porcine testis is understudied. Here, we performed a comprehensive analysis of the m6A transcriptome-wide profile in Shaziling pig testes at birth, puberty, and maturity. We analyzed the total transcriptome m6A profile and found that the m6A patterns were highly distinct in terms of the modification of the transcriptomes during porcine testis development. We found that key m6A methylated genes (AURKC, OVOL, SOX8, ACVR2A, and SPATA46) were highly enriched during spermatogenesis and identified in spermatogenesis-related KEGG pathways, including Wnt, cAMP, mTOR, AMPK, PI3K-Akt, and spliceosome. Our findings indicated that m6A methylations are involved in the complex yet well-organized post-transcriptional regulation of porcine testicular development and spermatogenesis. We found that the m6A eraser ALKBH5 negatively regulated the proliferation of immature porcine Sertoli cells. Furthermore, we proposed a novel mechanism of m6A modification during testicular development: ALKBH5 regulated the RNA methylation level and gene expression of SOX9 mRNA. In addition to serving as a potential target for improving boar reproduction, our findings contributed to the further understanding of the regulation of m6A modifications in male reproduction. © 2023 by the authors.

语种： 英文

作者： 颜赛娜;杨岸奇;唐湘薇;陈楚杰;尹艳飞;...

期刊： 畜牧与兽医,2023年55(09):1-9 ISSN：0529-5130

作者机构： 湖南农业大学动物科学技术学院,湖南 长沙 410128;畜禽遗传改良湖南省重点实验室,湖南 长沙 410128;[马佳佳; 唐湘薇; 颜赛娜; 冉茂良; 陈斌; 向娇娇; 杨岸奇; 尹艳飞; 陈楚杰] 湖南农业大学动物科学技术学院<&wdkj&>畜禽遗传改良湖南省重点实验室

关键词： SMAD6基因;支持细胞;增殖;凋亡

摘要： 睾丸未成熟支持细胞的增殖活性对种公畜繁殖性能有重要影响,睾丸基因组DNA甲基化(5mC)不同发育阶段特异性引起的基因差异表达可以调控支持细胞的增殖活性.在课题组前期研究中,SMAD6 被鉴定出是睾丸发育过程中受 3′UTR甲基化水平调控的枢纽基因之一,但其生物学功能尚未被发掘.本研究旨在探究SMAD6 对猪未成熟支持细胞增殖和凋亡的作用.在体外培养的猪未成熟支持细胞中过表达SMAD6 或干扰SMAD6 表达后,检测细胞周期和增殖相关基因的相对表达量、细胞增殖情况、细胞ATP水平、细胞凋亡率及凋亡相关蛋白的表达情况.结果显示:过表达SMAD6 后细胞周期及增殖相关基因的相对表达量上升(P＜0.05 或P＜0.01)、细胞增殖增加(P＜0.05 或P＜0.01)、细胞ATP水平升高(P＜0.01)、细胞的凋亡率下降(P＜0.05)、促凋亡相关蛋白BAX和Caspase3 的表达降低(P＜0.05);干扰SMAD6 表达结果则与之相反.研究结果提示,SMAD6 促进猪未成熟支持细胞的增殖且抑制其凋亡.

语种： 中文

作者： 楚丹;冉茂良;卞桥;熊锦翼;陈斌

期刊： 中国畜牧杂志,2022年58(4):77-83 ISSN：0258-7033

作者机构： [熊锦翼] 湖南农业大学动物科学技术学院;畜禽遗传改良湖南省重点实验室;[楚丹; 冉茂良; 陈斌; 卞桥] 湖南农业大学动物科学技术学院<&wdkj&>畜禽遗传改良湖南省重点实验室

关键词： 猪;进化;定量表达;靶基因

摘要： 为探究miRNA-139进化进程及其潜在靶基因功能，以及miR-139在猪睾丸发育过程中的表达差异和变化规律。本研究通过miRbase数据库获取全物种miR-139成熟序列，在Ensembl数据库中定位miR-139在多物种基因组中的位置；通过MEGA X软件选用邻近法构建关于miR-139的系统进化树；利用RT-qPCR实验技术分析miR-139在沙子岭猪7个时期（D1、D30、D60、D90、D120、D150和D180）的睾丸组织细胞中的差异表达及变化，并使用Targetscan等多个在线软件预测可能与猪miR-139结合的靶基因，对靶基因进行GO和KEGG分析。结果显示：miR-139基因在进化过程中具有较高的保守性；定量结果显示，miR-139在猪睾丸组织各7个时期有差异表达，1～90 d的表达水平逐渐降低，在性发育期120 d上升又在150 d有所下降，性成熟期表达水平又显著升高；miR-139靶基因GO和KEGG分析结果显示，miR-139的靶基因主要起结合、转录前调节的作用，多个基因富集在对畜禽繁殖和生长发育有重要调控作用的信号通路；4种在线软件取交集预测保守猪源候选靶基因有25个，为SCHIP1、WDR26、DMD、MIER3、DPYSL5、CANX、ASH1L、AJAP1等。本研究发现，miR-139在物种进化过程中高度保守，并对猪睾丸的生长发育有重要影响作用，以期为后续进一步探讨miR-139调控猪精子发生的过程提供理论基础。

语种： 中文

作者： 曾兴宇;唐湘薇;陈楚杰;楚丹;颜赛娜;...

期刊： 畜牧与兽医,2022年54(08):1-8 ISSN：0529-5130

作者机构： 湖南农业大学动物科学技术学院;畜禽遗传改良湖南省重点实验室;[唐湘薇; 楚丹; 颜赛娜; 冉茂良; 陈斌; 曾兴宇; 陈楚杰] 湖南农业大学动物科学技术学院<&wdkj&>畜禽遗传改良湖南省重点实验室

关键词： 支持细胞;增殖;凋亡

摘要： 支持细胞的增殖力决定性成熟后睾丸支持细胞的数量，进而影响公猪的精子发育、成熟及精液品质。microRNA通过结合靶基因的3′-UTR,广泛参与调控支持细胞增殖、凋亡和分化等多种生物学过程。为了研究miR-139在猪未成熟支持细胞中的作用，试验将miR-139过表达试剂、抑制表达试剂及真核翻译起始因子（EIF4G2）的干扰试剂分别转染到猪的未成熟支持细胞中，采用实时荧光定量及流式细胞术等方法对其进行研究。结果显示：过表达miR-139,细胞周期进程加快，增殖能力显著增强，而细胞凋亡率则显著降低；抑制表达miR-139后，结果与之相反。干扰EIF4G2基因后，细胞周期进程加快，且促进了细胞增殖而抑制其凋亡，与过表达miR-139的结果相一致。结果说明miR-139促进猪未成熟支持细胞增殖而抑制其凋亡，且其作用机制可能是通过靶向EIF4G2基因实现。该研究为进一步探讨支持细胞增殖对精子发生的作用提供理论依据。

语种： 中文

作者： Anqi, Yang;Saina, Yan;Chujie, Chen;Yanfei, Yin;Xiangwei, Tang;...

期刊： Genomics,2022年114(5):110450 ISSN：0888-7543

通讯作者： Maoliang, Ran;Bin, Chen

作者机构： [Jiaojiao, Xiang; Jiajia, Ma; Maoliang, Ran; Yanfei, Yin; Anqi, Yang; Saina, Yan; Xiangwei, Tang; Bin, Chen; Chujie, Chen] Hunan Agr Univ, Coll Anim Sci & Technol, Hunan Prov Key Lab Genet Improvement Domest Anim, Changsha 410128, Hunan, Peoples R China.

通讯机构： [Maoliang, Ran; Bin, Chen] C;College of Animal Science and Technology, Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Hunan, Changsha 410128, China. Electronic address:

关键词： DNA methylation;Transcriptome;WGBS;RNA-seq;Testis;Boars;INHBA

摘要： DNA methylation is one of the key epigenetic regulatory mechanisms in development and spermatogenesis. However, the dynamic regulatory mechanisms of genome-wide DNA methylation during testicular development remain largely unknown. Herein, we generated a single-base resolution DNA methylome and transcriptome atlas of precocious porcine testicular tissues across three developmental stages (1, 75, and 150 days old). The results showed that the dynamic methylation patterns were directly related to the expression of the DNMT3A gene. Conjoint analysis revealed a negative regulatory pattern between promoter methylation and the positive regulation of 3'-untranslated region (3'UTR) methylation. Mechanistically, the decrease in promoter methylation affected the upregulation of meiosis-related genes, such as HORMAD1, SPO11, and SYCE1. Demethylation in the 3'UTR induced the downregulation of the INHBA gene and knockdown of INHBA inhibited cell proliferation by reducing the synthesis of activin A. These findings contribute to exploring the regulatory mechanisms of DNA methylation in testicular development.

语种： 英文

作者： Chen, Chen;Zhu, Ji;Ren, Huibo;Deng, Yuan;Zhang, Xing;...

期刊： Livestock Science,2021年244:104342 ISSN：1871-1413

通讯作者： Maisheng Wu<&wdkj&>Yinglin Peng

作者机构： [Deng, Yuan; Chen, Chen; Hu, Xionggui; Zhu, Ji; Zhang, Xing; Cui, Qingming; Ren, Huibo; Liu, Yingying; Zuo, Jianbo; Peng, Yinglin] Hunan Inst Anim & Vet Sci, Changsha 410131, Hunan, Peoples R China.;[Deng, Yuan; Chen, Chen; Hu, Xionggui; Zhu, Ji; Zhang, Xing; Cui, Qingming; Ren, Huibo; Liu, Yingying; Zuo, Jianbo; Peng, Yinglin] Key Lab Conservat & Genet Anal Local Pig Breed Ge, Changsha 410131, Hunan, Peoples R China.;[Chen, Bin; Peng, Yinglin] Hunan Agr Univ, Coll Anim Sci & Technol, Changsha 410128, Hunan, Peoples R China.;[Zhang, Xing; Wu, Maisheng] Bur Agr & Rural Affairs Xiangtan City, Xiangtan 411104, Hunan, Peoples R China.

通讯机构： [Maisheng Wu] B;[Yinglin Peng] H;Bureau of Agriculture and Rural Affairs of Xiangtan City, Xiangtan, Hunan 411104, PR China<&wdkj&>Hunan Institute of Animal & Veterinary Science, Changsha, Hunan 410131, PR China<&wdkj&>Key Laboratory of Conservation and Genetic Analysis of Local Pig Breed Germplasm Resources, Changsha, Hunan 410131, PR China<&wdkj&>College of Animal Science and Technology, Hunan Agricultural University, Changsha, Hunan 410128, PR China

关键词： essential amino acid;monounsaturated fatty acid;saturated fatty acid;animal experiment;Article;biological trait;carcass;chemical composition;commercial phenomena;cross breeding;female;food industry;food quality;growth rate;male;nonhuman;pH;product quality;Yorkshire pig

摘要： The Shaziling pig, an indigenous breed reared in Hunan province in China, is characterized by good meat quality and strong ability of resistance for general diseases. The objective of this study was to compare properties of growth performance, carcass characteristics, meat quality, and chemical composition among purebred Shaziling, Berkshire, Yorkshire pigs and their crossbreeds Yorkshire × (Berkshire × Shaziling) (Y × (B × S)), Berkshire × (Yorkshire × Shaziling) (B × (Y × S)) pigs. The results showed that Shaziling pigs had lower average daily gain (ADG) (P < 0.01) and higher ratio of Feed/Gain (P < 0.01) than other groups. In contrast, the crossbreeds Y × (B × S) and B × (Y × S) both exhibited faster growth rate and superior carcass characteristics. In particular, the growth performance and carcass traits of Y × (B × S) pigs were very close to the lean-type Berkshire and Yorkshire pigs, including ADG, dressing percentage, loin-eye area and lean percentage. Meanwhile, the parameters associated with meat quality were assigned a normal quality class in all breeds, such as pH, meat color, and drip loss. Furthermore, both crossbreeds had desirable chemical composition, as demonstrated by moderate IMF and abundant mineral elements, SFAs, MUFAs, EAAs, and FAAs. Taken together, the crossbreed of Y × (B × S) is preferably suited for commercial production to provide high tasted quality niche products. © 2020 Elsevier B.V.

语种： 英文

作者： 张仙玉;赵鑫;李国玲;邢萍萍;李紫聪;...

期刊： 畜牧兽医学报,2021年52(02):408-419 ISSN：0366-6964

作者机构： [陈斌] 湖南农业大学动物科学技术学院;[吴珍芳; 杨化强; 赵鑫; 李国玲; 邢萍萍; 李紫聪] 华南农业大学动物科学学院国家生猪种业工程技术研究中心;[张仙玉] 湖南农业大学动物科学技术学院<&wdkj&>华南农业大学动物科学学院国家生猪种业工程技术研究中心

关键词： 体外培养;细胞增殖;功能鉴定

摘要： 旨在通过探索ICR品系小鼠精原干细胞（spermatogonial stem cells, SSCs）的体外稳定培养,为别的品系小鼠和大动物的SSCs体外培养提供参考。本研究采集6～8日龄ICR乳鼠的睾丸,用两步酶消化法（胶原酶和胰蛋白酶）和差速贴壁法来纯化SSC细胞。用不同的饲养层（小鼠胎儿成纤维细胞（mouse embryonic fibroblasts, mEF）作为饲养层和层黏连蛋白与多聚赖氨酸联合铺板）,不同的培养基（StemPro-34培养基和DMEM培养基）,添加不同的生长因子（GDNF、LIF、EGF、bFGF、IGF1）来观察体外SSCs的生长。将传代至第6代的SSCs用免疫染色法和分子检测法来评估细胞增殖。最后将稳定增殖的SSCs移植到受体睾丸中进行功能鉴定。结果显示,通过两步酶消化法和差速贴壁法获得纯度约79%以上的SSCs细胞;mEF作为饲养层,StemPro-34作为培养基,组合添加GDNF、LIF、EGF、bFGF、IGF1生长因子能促进SSCs的体外长期培养;SSCs克隆集落用碱性磷酸酶（alkaline phosphatase, AKP）染色、PLZF和UCHL1蛋白免疫荧光检测均显示为阳性,用反转录聚合酶链反应（reverse transcription-polymerase chain reaction, RT-PCR）和荧光实时定量PCR（quantitative real-time PCR, qRT-PCR）检测显示,克隆集落高表达多能基因和自我更新基因,说明体外培养的SSCs实现了稳定增殖;SSCs（带有报告基因EGFP）移植到受体睾丸后,受体附睾中精子头部发出绿光,证明精子为供体SSCs来源。研究表明,本试验中的培养体系适用于ICR品系小鼠的体外培养,且稳定培养后的SSCs具有正常生物学功能。本研究为别的品系小鼠和大动物SSCs的体外培养提供了参考和借鉴。

语种： 中文

作者： 唐湘薇;高虎;陈斌

期刊： 经济动物学报,2021年25(02):120-124 ISSN：1007-7448

作者机构： 湖南农业大学动物科学技术学院;畜禽遗传改良湖南省重点实验室;[唐湘薇; 高虎; 陈斌] 湖南农业大学动物科学技术学院<&wdkj&>畜禽遗传改良湖南省重点实验室

关键词： 环状RNA;生物学功能;高通量测序

摘要： 环状RNA（circRNA）是一种广泛、稳定存在于哺乳动物中的不具备5’末端帽子和3’末端poly（A）尾巴且具有独特闭环结构的内源性非编码RNA,具有高度保守性、组织特异性、阶段特异性以及高度稳定性等优点。按形成方式主要分为内含子circRNA、外显子circRNA、外显子-内含子circRNA以及基因间circRNA。目前对其功能的研究主要集中在与miRNA的互作、蛋白调控以及癌症等疾病方面。文中就circRNA的生物合成、功能及在畜牧学中的研究进展进行综述,以期为畜牧分子育种研究提供理论支撑。

语种： 中文

作者： Zhang, Xianyu;Zhao, Xin;Li, Guoling;Zhang, Mao;Xing, Pingping;...

期刊： BIOLOGY OPEN,2021年10(1):bio056804 ISSN：2046-6390

通讯作者： Chen, Bin;Yang, Huaqiang;Wu, Zhenfang

作者机构： [Chen, Bin; Zhang, Xianyu] Hunan Agr Univ, Coll Anim Sci & Technol, Changsha 410128, Hunan, Peoples R China.;[Xing, Pingping; Li, Zicong; Zhao, Xin; Zhang, Mao; Wu, Zhenfang; Yang, Huaqiang; Li, Guoling; Zhang, Xianyu] South China Agr Univ, Natl Engn Res Ctr Breeding Swine Ind, Guangzhou 510642, Peoples R China.

通讯机构： [Chen, Bin] H;[Yang, HQ; Wu, ZF] S;Hunan Agr Univ, Coll Anim Sci & Technol, Changsha 410128, Hunan, Peoples R China.;South China Agr Univ, Natl Engn Res Ctr Breeding Swine Ind, Guangzhou 510642, Peoples R China.

关键词： Etv5;Spermatogonial stem cells;Gene knockout;Transplantation;Spermatogenesis

摘要： Spermatogonial stem cell (SSC) transplantation is an alternative reproductive method to achieve conservation and production of elite animals in livestock production. Creating a recipient animal without endogenous germ cells is important for effective SSC transplantation. However, natural mutants with depletion of SSCs are difficult to obtain, and drug ablation of endogenous germ cells is arduous to perform for practical use. In this study, we used mouse models to study the preparation of recipients with congenital germ cell ablation. We knocked out (KO) Ets-variant gene 5 (Etv5) in mice using the CRISPR/Cas9 system. The testicular weight of Etv5−/− mice was significantly lower than that of wild-type (WT) mice. The germ cell layer of the seminiferous tubules gradually receded with age in Etv5−/− mice. At 12 weeks of age, the tubules of Etv5−/− mice lacked almost all spermatogenic cells with a Sertoli cell-only phenotype, and sperm were completely absent in the epididymis. We subsequently transplanted allogeneic SSCs with enhanced green fluorescent protein (EGFP) into 3- (immature) or 7-week-old (mature) Etv5−/− mice. Partially restoration of germ cell layers in the seminiferous tubules and spermatogenesis was observed in all immature testes but not in mature adult testes at 2 months post-transplantation. The presence of heterologous genes Etv5 and EGFP in recipient testicular tissue and epididymal sperm by PCR indicated that sperm originated from the transplanted donor cells. Our study demonstrates that, although Etv5−/− mice could accommodate and support foreign germ cell transplantation, this process occurs in a quite low efficiency to support a full spermatogenesis of transplanted SSCs. However, using Etv5−/− mice as a recipient model for SSC transplantation is feasible, and still needs further investigation to establish an optimized transplantation process. © 2021. Published by The Company of Biologists Ltd

语种： 英文

作者： 刘思远;易国强;唐中林;陈斌

期刊： 遗传,2020年42(5):435-443 ISSN：0253-9772

作者机构： [刘思远] 湖南农业大学动物科学技术学院,长沙 410128;[唐中林; 易国强; 刘思远] 中国农业科学院农业基因组研究所,岭南现代农业科学与技术广东省实验室深圳分中心,农业部农业基因数据分析重点实验室,深圳 518120;[唐中林] 中国农业科学院北京畜牧兽医研究所,北京 100193;[陈斌] 湖南农业大学

关键词： 全基因组筛选;功能基因;调控元件

摘要： CRISPR/Cas9系统是一种近年来被广泛应用于基因组编辑的强大工具。通过将CRISPR/Cas9系统中的Cas9蛋白突变后,使其失去剪切活性而成为dCas9 (nuclease-dead Cas9),再结合基因功能丧失(loss-of-function, LOF)、基因功能激活(gain-of-function, GOF)以及非编码功能基因鉴定技术即可实现全基因组高通量的功能基因及调控元件靶向鉴定和筛选。目前,该技术已被广泛应用于疾病免疫机理、药物靶点筛选和动物遗传育种等研究,为生命医学和基础科学带来了全新高效的技术方法和研究思路。本文综述了基于CRISPR/Cas9技术在全基因组中高通量筛选功能基因及调控元件的方法及研究进展,重点阐述了CRISPR/Cas9系统在动物细胞中筛选功能性基因的方法,以期为基因编辑及相关研究领域提供参考。

语种： 中文

作者： 陈晨;张星;罗璇;邓缘;崔清明;...

期刊： 湖南农业大学学报(自然科学版),2020年46(3):343-349 ISSN：1007-1032

作者机构： 湖南省畜牧兽医研究所,湖南长沙410131;湘潭市农业农村局,湖南湘潭411104;湖南农业大学动物科学技术学院,湖南长沙410128;[彭英林; 罗璇; 崔清明; 陈晨; 张星; 邓缘] 湖南省畜牧兽医研究所;[陈斌] 湖南农业大学

关键词： 大巴沙猪;屠宰体质量;胴体性状;肌肉品质

摘要： 以湘沙猪配套系商品猪大巴沙猪为研究对象,探讨不同屠宰体质量阶段(85~95、>95~105、>105~115 kg)对胴体性能和肌肉品质的影响,并分析性状间的相关性。结果表明:胴体性状方面,85~95 kg大巴沙猪的平均背膘厚最薄,腿臀比例最高,>95~105 kg大巴沙猪的瘦肉率最高,>105~115 kg大巴沙猪的屠宰率最高,胴体最长,眼肌面积最大;肉质性状方面,85~95 kg大巴沙猪的肌肉水分含量最高,肌内脂肪含量最低,>95~105 kg大巴沙猪的熟肉率、多不饱和脂肪酸含量最高,饱和脂肪酸含量最低,>105~115 kg大巴沙猪的系水力、肌内脂肪、肌苷酸、饱和脂肪酸和单不饱和脂肪酸含量最高,滴水损失、多不饱和脂肪酸含量最低;所有被测猪只的肌肉必需氨基酸,风味氨基酸和微量元素含量均很丰富;根据拟合优度建立了部分性状间的回归方程。综合分析,>105~115 kg大巴沙猪的胴体性状佳、肌肉品质好,>105~115 kg为适宜上市屠宰体质量。

语种： 中文

作者： 张兴;陈晨;谭红;李玉莲;彭英林;...

期刊： 养猪,2020年(02):65-68 ISSN：1002-1957

作者机构： 湘潭市家畜育种站,湖南 湘潭 411104;湖南省畜牧兽医研究所,湖南 长沙 410131;湖南农业大学动物科学技术学院,湖南 长沙 410126;湘潭市农业农村局,湖南 湘潭 411104;[彭英林; 陈晨] 湖南省畜牧兽医研究所

关键词： XS3系;XS2系;XS1系;胴体品质;肉质性状;必需氨基酸评分

摘要： 为了比较湘沙猪配套系3个专门化品系(XS3系、XS2系和XS1系)胴体品质、肉质性状、必需氨基酸评分等方面差异,试验分别选择6头、6头、5头体重为90 kg、95 kg、115 kg左右的健康XS3系、XS2系、XS1系,按品系分为3组,进行屠宰测定,比较分析相关指标。结果表明:1)XS3系的3点平均背膘厚、脂率显著高于XS1系、XS2系(P<0.05),胴体斜长、瘦肉率、3点平均皮厚、皮率3组间均差异显著(P<0.05),XS3系的眼肌面积、后腿比例、骨率显著低于XS2系、XS1系(P<0.05);2)XS2系的背最长肌pH 1 h、b*1 h显著高于XS1系、XS3系(P<0.05),XS1系的背最长肌pH 24 h显著低于XS2系、XS3系(P<0.05),XS3系的L^*1 h显著高于XS1(P<0.05),XS2系的a^*1 h显著高于XS1系(P<0.05),XS3系、XS2系的肉色等级、大理石纹显著高于XS1系(P<0.05);3)XS2系的水分含量显著高于XS3系(P<0.05),XS3系的粗脂肪含量显著高于XS2系和XS1系(P<0.05);4)3组的必需氨基酸得分均差异不大。说明湘沙猪配套系3个专门化品系,在胴体品质上以XS1系最优,XS2系居中,XS3系次之;在肉质性状上XS3系最优、XS2系居中、XS1系次之;必需氨基酸评分无明显区别。

语种： 中文

作者： 杨岸奇;陈斌;冉茂良;杨广民;曾诚

期刊： 遗传,2020年42(2):145-152 ISSN：0253-9772

作者机构： [杨岸奇] 湖南农业大学动物科学技术学院,长沙 410128;[杨岸奇] 湖南美可达生物资源股份有限公司,长沙 410331;[杨岸奇] 畜禽遗传改良湖南省重点实验室,长沙 410128;[杨广民; 曾诚] 湖南美可达生物资源股份有限公司;[冉茂良; 陈斌] 畜禽遗传改良湖南省重点实验室

关键词： 基因组选择;猪;杂交育种;统计模型

摘要： 基因组选择是指在全基因组范围内通过基因组中大量的标记信息估计出个体全基因组范围的育种值,可进一步提升育种效率和准确性,目前在猪纯繁育种中得到广泛应用。但有研究表明,现有的基因组选择方法在猪杂交育种上的应用效果并不理想,在跨群体条件下预测准确性极低。杂交作为养猪业中最为广泛的育种手段之一,通过结合基因组选择理论进一步提升猪的生产性能,具有重要的经济和研究价值。本文综述了基因组选择的发展及其在猪育种中的应用现状,并结合国内外猪杂交育种的方式,分析了目前基因组选择方法在猪杂交育种应用方面的不足,旨在为未来基因组选择在猪杂交育种中的合理应用提供参考。

语种： 中文

作者： 石兴勇;王吉;陈斌

期刊： 中国实验动物学报,2019年27(1):104-109 ISSN：1005-4847

作者机构： [石兴勇] 湖南农业大学 动物科学技术学院,长沙 410000;[石兴勇] 湖南赛诺生物科技股份有限公司,长沙 410000;湖南农业大学 动物医学院,长沙 410000;长沙绿叶生物科技有限公司,长沙 410000;湖南农业大学 动物科学技术学院,长沙,410000

关键词： 实验动物;小型猪;资源;品系

摘要： 实验动物广泛应用于科学研究领域,实验小型猪在医学研究领域中越来越得到重视。中国小型猪种源丰富,不同种群各具特色,遗传性状稳定。近年来中国实验小型猪的研究处于快速发展阶段,生产和使用均在逐年增加。我国发展实验小型猪具有天然封闭群特性和高度近交等得天独厚的条件,品系化、标准化、规模化和产业化是我国实验用小型猪的主要发展趋势。我国实验用小型猪发展当前面临的问题主要表现在:保护力度小、发展阻力大、育成品系少三个方面。育成中国各具特色的实验用小型猪品系需要增加投入,利用先进的科学技术例如基因编辑和动物克隆技术,加速小型猪的定向化培育进程。

语种： 中文

作者： 陈晨;邓缘;胡雄贵;刘莹莹;吴买生;...

期刊： 家畜生态学报,2019年40(1):13-17 ISSN：1673-1182

作者机构： 湖南省畜牧兽医研究所地方猪种质资源保护与遗传解析湖南省重点实验室;[陈斌] 湖南畜禽安全生产协同创新中心;[吴买生] 湘潭市畜牧兽医水产局;湖南农业大学动物科学技术学院;[彭英林; 罗璇; 刘莹莹; 陈晨; 胡雄贵; 张星; 邓缘] 湖南省畜牧兽医研究所

关键词： 猪;肌苷酸含量;基因表达;相关性分析

摘要： 以沙子岭猪、大约克、巴克夏猪及其杂交组合巴×沙、大×巴沙猪为研究对象,测定背最长肌中肌苷酸含量,对ADSL、GPAT和PURH基因在不同群体背最长肌中的表达水平进行检测,分析其与肌苷酸含量的相关性.结果表明,沙子岭猪肌苷酸含量显著高于巴克夏和大×巴沙猪(P＜0.05),其余各群体之间差异均不显著(P＞0.05).肌苷酸含量与肌肉pH1 h、肉色L*值和a*值、肌内脂肪含量、剪切力等肉质指标具有显著相关性.ADSL、GPAT和PURH基因在不同群体中的表达模式基本相同,即三种基因均在沙子岭猪和巴×沙猪中高表达.ADSL基因表达量与肌苷酸含量呈极显著正相关(P＜0.01),GPAT和PURH基因表达量与肌苷酸含量均呈显著正相关(P＜0.05).结果提示,ADSL、GPAT和PURH基因与肌苷酸沉积紧密相关,或可作为猪肉质风味的选择依据,为今后选育工作提供理论基础.

语种： 中文

作者： 高虎;冉茂良;翁波;彭馥芝;罗荟;...

期刊： 经济动物学报,2019年23(02):94-101+106 ISSN：1007-7448

作者机构： 湖南农业大学动物科学技术学院,长沙410128;畜禽遗传改良湖南省重点实验室,长沙410128;[冉茂良; 陈斌; 罗荟; 翁波; 彭馥芝; 高虎] 畜禽遗传改良湖南省重点实验室

关键词： 猪;进化;定量表达

摘要： 采用生物学信息方法在miRBase中搜索动物miR-125a/125b基因家族序列,利用Ensembl数据库信息确定miR-125a/125b在基因组中的位置,采用MEGA 5.0构建miR-125a/125b家族的系统进化树,并利用qRT-PCR检测miR-125a和miR-125b在沙子岭猪睾丸组织不同发育时期(D1、D30、D60、D90、D120、D150、D180)的表达变化.结果在miRBase数据库检索29个物种共58条miR-125家族序列,分布广泛,且有高度的保守性,除一个位于内含子区,两个位于外显子区,其余miR-125a/125b位于基因间隔区,在进化过程中,同种物种的进化方向具有高度的一致性,进化速度也有所不同.qRT-PCR检测结果显示,miR-125a/125b在沙子岭猪睾丸组织中不同发育时期的具有相同的趋势,即出生后其表达水平缓慢升高,保育期开始降低,性成熟期又开始呈现升高趋势,随着体成熟表达量开始降低.本研究结果为进一步探明miR-125a/125b基因家族在猪睾丸组织中的功能及作用机制奠定基础.

语种： 中文

作者： Weng Bo;Ran Mao-liang;Cao Rong;Peng Fu-zhi;Luo Hui;...

期刊： 农业科学学报(英文),2019年18(8):1924-1935 ISSN：2095-3119

通讯作者： Chen Bin

作者机构： [Yang An-qi; Weng Bo; Ran Mao-liang; Chen Bin; Gao Hu; Cao Rong; Tang Xiang-wei; Peng Fu-zhi; Luo Hui] Hunan Agr Univ, Hunan Prov Key Lab Genet Improvement Domest Anim, Coll Anim Sci & Technol, Changsha 410128, Hunan, Peoples R China.

通讯机构： [Chen Bin] H;Hunan Agr Univ, Hunan Prov Key Lab Genet Improvement Domest Anim, Coll Anim Sci & Technol, Changsha 410128, Hunan, Peoples R China.

关键词： miR-10b;DAZAP1;gene expression;proliferation;porcine immature Sertoli cell

摘要： MicroRNAs (miRNAs) have been widely identified in porcine testicular tissues and implicated as crucial regulators of proliferation, apoptosis, and differentiation in porcine spermatogenesis related cells. However, the function roles of most of the miRNAs that have been identified in Sertoli cells are poorly understood. In the present study, six experiments were conducted to study the regulatory role of miR-10b in porcine immature Sertoli cells. In experiment 1, the results showed that the relative mRNA expression level of miR-10b in porcine testicular tissues decreased quadratically (P<0.001) with increasing age, while the relative mRNA expression level of DAZAP1 gene increased (P<0.001). In addition, the mRNA expression of miR-10b was negatively (P<0.01) correlated with DAZAP1 mRNA expression (r=−0.550). In experiment 2, the results from the bioinformatic analysis and a luciferase reporter assay demonstrated that miR-10b directly targeted the DAZAP1 gene in porcine immature Sertoli cells. DAZAP1 mRNA and protein expressions were both regulated (P<0.05) by miR-10b. In experiments 3 to 5, the over-expression of miR-10b or the siRNA-mediated knockdown of the DAZAP1 gene promoted (P<0.05) porcine immature Sertoli cell proliferation, as determined by the Cell Counting Kit-8 (CCK-8) assay and the 5-Ethynyl-2'-deoxyuridine (EdU) assay. However, an annexin V-FITC/PI staining assay and the expression of cell survival-related genes indicated that over-expression of miR-10b or knockdown of DAZAP1 had no effect (P>0.05) on porcine immature Sertoli cell apoptosis. In experiment 6, the co-transfection treatment results showed that miR-10b promoted (P<0.05) porcine immature Sertoli cell proliferation by targeting DAZAP1 gene. Overall, these experiments demonstrated that miR-10b promotes porcine immature Sertoli cell proliferation by targeting the DAZAP1 gene. © 2019 CAAS. Publishing services by Elsevier B.V

语种： 英文

作者： 楚丹;陈斌;冉茂良

期刊： 中国猪业,2019年14(6):11-15 ISSN：1673-4645

作者机构： 湖南农业大学动物科学技术学院;[陈斌] 畜禽遗传改良湖南省重点实验室;[楚丹] 湖南农业大学

关键词： 胎次;配种月份;繁殖性能;长白二元母猪

摘要： 胎次和配种月份等非遗传因素对母猪繁殖性能具有重要的影响.本文于2010年1月至2017年12月对湖南某猪场共计3636头长白二元母猪的总产仔数、 产活仔数、 畸形数、 木乃伊数、 死胎数、 弱仔数、 死仔数和初生窝重进行统计分析,研究胎次和配种月份对长白二元母猪繁殖性能的影响.结果显示,胎次和配种月份对长白二元母猪繁殖性能有显著影响;从第1胎至第3胎母猪的总产仔数、 产活仔数、 初生窝重呈逐渐递增趋势,其中,第3胎次最高,3～5胎次繁殖性能趋于稳定;第1胎的死胎数、 畸形数、 弱仔数最高;第5胎的木乃伊数最高,但畸形数最低.长白二元母猪在每年2—4月的配种繁殖性能最佳,总产仔数、 产活仔数显著高于5—7月.结果表明,长白母猪第2～5胎次繁殖性能较好,且适宜配种月份为2—4月.

语种： 中文

作者： Ran Mao-liang;Weng Bo;Cao Rong;Peng Fu-zhi;Luo Hui;...

期刊： 农业科学学报(英文),2019年18(2):449-459 ISSN：2095-3119

通讯作者： Chen Bin

作者机构： [Weng Bo; Ran Mao-liang; Chen Bin; Gao Hu; Cao Rong; Peng Fu-zhi; Luo Hui] Hunan Agr Univ, Hunan Prov Key Lab Genet improvement Domest Anim, Coll Anim Sci & Technol, Changsha 410128, Hunan, Peoples R China.

通讯机构： [Chen Bin] H;Hunan Agr Univ, Hunan Prov Key Lab Genet improvement Domest Anim, Coll Anim Sci & Technol, Changsha 410128, Hunan, Peoples R China.

关键词： immature porcine Sertoli cell;cell proliferation;apoptosis;SMAD7;miR-34c

摘要： MicroRNAs (miRNAs) are implicated in swine spermatogenesis via their regulations of cell proliferation, apoptosis, and differentiation. Recent studies indicated that miR-34c is indispensable in the late steps of spermatogenesis. However, whether miR-34c plays similar important roles in immature porcine Sertoli cells remain unknown. In the present study, we conducted two experiments using a completely randomised design to study the function roles of miR-34c. The results from experiment I demonstrated that the relative expression level of miR-34c in swine testicular tissues increased (P=0.0017) quadratically with increasing age, while the relative expression level of SMAD family member 7 (SMAD7) decreased (P=0.0009) with curve. Furthermore, miR-34c expression levels showed a significant negative correlation (P=0.013) with SMAD7 gene expression levels. The results from experiment II indicated that miR-34c directly targets the SMAD7 gene using a luciferase reporter assay, and suppresses (P<0.05) SMAD7 mRNA and protein expressions in immature porcine Sertoli cells. Overexpression of miR-34c inhibited (P<0.05) proliferation and enhanced (P<0.05) apoptosis in the immature porcine Sertoli cells, which was supported by the results from the Cell Counting Kit-8 (CCK-8) assay, the 5-Ethynyl-2'-deoxyuridine (EdU) assay, and the Annexin V-FITC/PI staining assay. Furthermore, knockdown of SMAD7 via small interfering RNA (siRNA) gave a similar result. It is concluded that miR-34c inhibits proliferation and enhances apoptosis in immature porcine Sertoli cells by targeting the SMAD7 gene.

语种： 英文