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Meclofenamic acid represses spermatogonial proliferation through modulating m^6A RNA modification

Journal of animal Science and Biotechnology 2019年第4期10卷 919-930页

作者： Tao Huang Jiayin Guo Yinghua Lv Yi Zheng Tongying Feng Qiang Gao Wenxian Zeng key laboratory of animal genetics Breeding and Reproduction of Shaanxi Province College of Animal Science and Technology Northwest A&F University

Background: N6-Methyladenosine(m^6A), the most prevalent modification in mammalian m RNA, plays important roles in numerous biological processes. Several m^6A associated proteins such as methyltransferase like 3(METTL3),methyltransferase like 14(METTL14), α-ketoglutarate-dependent dioxygenase Alk B homolog 5(ALKBH5) and YTH domain containing 2(YTHDC2) are involved in the regulation of spermatogenesis and oogenesis. However, the role of the first detected m^6A demethylase, fat mass and obesity associate protein(FTO), in germ cells remains *** of FTO roles in the regulation of germ cell fate will provide novel insights into the mammalian ***: Mouse GC-1 spg cells were treated with the ester form of meclofenamic acid(MA2) to inhibit the demethylase activity of FTO. The cellular m^6A and m^6Amlevel were analyzed through high performance liquid chromatography combined with tandem mass spectrometry(HPLC/MS-MS). The cell apoptosis was detected via TUNEL and flow cytometry. The cell proliferation was detected through Ed U and western blot. The m RNA level of core cyclin dependent kinases(CDKs) was quantified via q-PCR. RNA decay assay were performed to detect RNA stability. Dual fluorescence assay was conducted to study whether MA2 affects the expression of CDK2 dependent on the m^6A modification at 3’***: MA2 significantly increased the cellular m^6A level and down-regulated the expression of CDK1, CDK2, CDK6 and Cd C25 a, resulting in arrest of G1/S transition and decrease of cell proliferation. MA2 downregulated CDK2 m RNA stability. Additionally, mutation of the predicted m^6A sites in the Cdk2–3’UTR could mitigated the degradation of CDK2 m RNA after MA2 ***: MA2 affected CDKs expression through the m^6A-dependent m RNA degradation pathway, and thus repressed spermatogonial proliferation.

关键词： Cell cycle FTO Meclofenamic acid N6-methyladenosine Spermatogonial proliferation

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Genomic analyses reveal distinct genetic architectures and selective pressures in Chinese donkeys

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Journal of genetics and Genomics 2021年第8期48卷 737-745页

作者： Jiafei Shen Jie Yu Xuelei Da Mei Li Gang Wang Ningbo Chen Hong Chen Chuzhao Lei Ruihua Dang key laboratory of animal genetics Breeding and Reproduction of Shaanxi ProvinceCollege of Animal Science and TechnologyNorthwest A&F UniversityYangtingShaanxi 712100China

Donkey(Equus asinus)is an important livestock animal in China because of its draft and medicinal *** a long period of natural and artificial selection,the variety and phenotype of donkeys have become *** clarified the genetic and demographic characteristics of Chinese domestic donkeys and the selection pressures by analyzing 78 whole genomes from 12 *** to population structure,most Chinese domestic donkeys showed a dominant ancestral ***,the Chinese donkeys still represented a significant geographical distribution *** the selective sweep,gene annotation,functional enrichment,and differential expression analyses between large and small donkey groups,we identified selective signals,including NCAPG and LCORL,which are related to rapid growth and large body *** findings elucidate the evolutionary history and formation of different donkey breeds and provide theoretical insights into the genetic mechanism underlying breed characteristics and molecular breeding programs of donkey clades.

关键词： Donkey Whole-genome resequencing NCAPG LCORL

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Adipose-derived stem cells and obesity: The spear and shield relationship

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Genes & Diseases 2023年第1期10卷 175-186页

作者： Hong Yang Chaowei Li Yizhou Li Ruiqing Tai Chao Sun key laboratory of animal genetics Breeding and Reproduction of Shaanxi ProvinceCollege of Animal Science and TechnologyNorthwest A&F UniversityYanglingShaanxi 712100China

With the transformation of modern lifestyles and population ageing,obesity has become a global epidemic,as one of the important threat to human health of chronic noncommunicable diseases(NCD).Stem cell therapy seems promising as an alternative strategy for managing obesity and related metabolic *** tissue-derived stem cells(ADSCs)have received widespread attention,which provides new ideas for the treatment of obesity and various metabolic-related diseases,due to their abundant reserves,easy acquisition,rapid expansion,and multi-directional differentiation potential,low immunogenicity and many other ***,there seems to be a"shield and spear paradox"in the relationship between ADsCs and *** this review,we emphatically summarized the role of ADsCs in the occurrence and development of obesity and related metabolic disease processes,in order to pave the way for clinical practice.

关键词： Adipose-derived stem cells Differentiation Inflammation Insulin resistance Metabolic syndrome Obesity

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Establishment of cell lines with porcine spermatogonial stem cell properties

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Journal of animal Science and Biotechnology 2020年第3期11卷 678-689页

作者： Yi Zheng Tongying Feng Pengfei Zhang Peipei Lei Fuyuan Li Wenxian Zeng key laboratory for animal genetics Breeding and Reproduction of Shaanxi ProvinceCollege of Animal Science and TechnologyNorthwest A&F UniversityYangling 712100ShaanxiChina

Background:Spermatogonial stem cells(SSCs)are capable of both self-renewal and differentiation to mature functional spermatozoa,being the only adult stem cells in the males that can transmit genetic information to the next *** SSCs hold great value in transgenic pig production and in establishment of porcine models for regenerative ***,studies and applications of porcine SSCs have been greatly hampered by the low number of SSCs in the testis as well as the lack of an ideal stable long-term culture system to propagate porcine SSCs ***:In the present study,by lentiviral transduction of plasmids expressing the simian virus 40(SV40)large T antigen into porcine primary SSCs,we developed two immortalized cell lines with porcine SSC *** established cell lines,with the expression of porcine SSC and germ cell markers UCHL1,PLZF,THY1,VASA and DAZL,could respond to retinoic acid(RA),and could colonize the recipient mouse testis without tumor formation after *** cell lines displayed infinite proliferation potential,and have now been cultured for more than 7 months and passaged for over 35 times without morphological ***:We have for the first time established porcine SSC lines that could provide abundant cell sources for mechanistic studies on porcine SSC self-renewal and differentiation,thereby facilitating development of an optimal long-term culture system for porcine primary SSCs and their application to animal husbandry and medicine.

关键词： Immortalization Pig Self-renewal Spermatogonial stem cells SV40 large T antigen

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circSVIL regulates bovine myoblast development by inhibiting STAT1 phosphorylation

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Science China(Life Sciences) 2022年第2期65卷 376-386页

作者： Binglin Yue Haiyan Yang Jiyao Wu Jian Wang Wenxiu Ru Jie Cheng Yongzheng Huang Xianyong Lan Chuzhao Lei Hong Chen key laboratory of animal genetics Breeding and Reproduction of Shaanxi ProvinceCollege of Animal Science and TechnologyNorthwest A&F UniversityYangling 712100China

Circular RNAs(circRNAs),a novel class of non-coding RNAs with a loop structure,have recently been shown to participate in various pathophysiological ***,the precise role of circRNAs in myoblasts remains *** this report,circSVIL was screened and identified from our previous sequencing analysis;we then performed gain-and loss-of-function experiments on bovine myoblasts by CCK8,EdU,flow cytometry,qRT-PCR,and Western *** results indicate that circSVIL facilitates bovine myoblast proliferation and inhibits cell *** mechanism assays such as bioinformatics prediction,RNA immunoprecipitation(RIP),and cytoplasmic separation,we demonstrate that circSVIL could interact with STAT1 and inhibit STAT1 phosphorylation,thereby restraining STAT1’s nuclear translocation and affecting its downstream signal *** results may elucidate a new regulatory pathway for bovine skeletal muscle development.

关键词： circSVIL STAT1 skeletal muscle

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Recent advances in isolation,identification,and culture of mammalian spermatogonial stem cells

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Asian Journal of Andrology 2022年第1期24卷 5-14页

作者： Hua-Ming Xi Yi-Jie Ren Fa Ren Yu Li Tian-Yu Feng Zhi Wang Ye-Qing Du Li-Kun Zhang Jian-Hong Hu key laboratory of animal genetics Breeding and Reproduction of Shaanxi ProvinceCollege of Animal Science and TechnologyNorthwest Agriculture and Forestry UniversityYangling 712100China

Continuous spermatogenesis depends on the self-renewal and differentiation of spermatogonial stem cells(SSCs).SSCs,the only male reproductive stem cells that transmit genetic material to subsequent generations,possess an inherent self-renewal ability,which allows the maintenance of a steady stem cell *** eventually differentiate to produce ***,in an in vitro culture system,SSCs can be induced to differentiate into various types of germ *** SSCs are well defined,and a culture system has been successfully established for *** contrast,available information on the biomolecular markers and a culture system for livestock SSCs is *** review summarizes the existing knowledge and research progress regarding mammalian SSCs to determine the mammalian spermatogenic process,the biology and niche of SSCs,the isolation and culture systems of SSCs,and the biomolecular markers and identification of *** information can be used for the effective utilization of SSCs in reproductive technologies for large livestock animals,enhancement of human male fertility,reproductive medicine,and protection of endangered species.

关键词： culture identification isolation male germline stem cell spermatogonial stem cell transplantation

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Single‑cell sequencing reveals the reproductive variations between primiparous and multiparous Hu ewes

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Journal of animal Science and Biotechnology 2024年第2期15卷 614-631页

作者： Ting Ge Yifan Wen Bo Li Xiaoyu Huang Shaohua Jiang Enping Zhang key laboratory of animal genetics Breeding and Reproduction of Shaanxi ProvinceCollege of Animal Science and TechnologyNorthwest A&F UniversityYangling 712100China

Background In the modern sheep production systems,the reproductive performance of ewes determines the economic profitability of *** the genetic mechanisms underlying differences in the litter size is important for the selection and breeding of highly prolific *** sheep,a high-quality Chinese sheep breed,is known for its high fecundity and is often used as a model to study prolificacy *** the current study,animals were divided into two groups according to their delivery rates in three consecutive lambing seasons(namely,the high and low reproductive groups with≥3 lambs and one lamb per season,n=3,respectively).The ewes were slaughtered within 12 h of estrus,and unilateral ovarian tissues were collected and analyzed by 10×Genomics single-cell RNA *** A total of 5 types of somatic cells were identified and corresponding expression profiles were mapped in the ovaries of each ***,the differences in the ovary somatic cell expression profiles between the high and low reproductive groups were mainly clustered in the granulosa ***,four granulosa cell subtypes were *** analysis revealed that the abundance of JPH1 expression and the reduction of LOC101112291 expression could lead to different evolutionary directions of the granulosa ***,the expression levels of FTH1 and FTL in mural granulosa cells of the highly reproductive group were significantly *** genes inhibit necroptosis and ferroptosis of mural granulosa cells,which helps prevent follicular *** This study provides insights into the molecular mechanisms underlying the high fecundity of Hu *** differences in gene expression profiles,particularly in the granulosa cells,suggest that these cells play a critical role in female *** findings also highlight the importance of genes such as JPH1,LOC101112291,FTH1,and FTL in regulating granulosa cell function and follicular development.

关键词： Granulosa cells Hu sheep Lambing number Ovarian somatic cells Single-cell RNA sequencing

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Multi-omics revealed the mechanism of feed efficiency in sheep by the combined action of the host and rumen microbiota

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animal Nutrition 2024年第3期18卷 367-379页

作者： Guangchen Zhou Junda Li Xuhui Liang Bohua Yang Ximeng He Hongyu Tang Hongran Guo Gongwei Liu Wenyuan Cui Yulin Chen Yuxin Yang key laboratory of animal genetics Breeding and Reproduction of Shaanxi ProvinceCollege of Animal Science and TechnologyNorthwest A&F UniversityYangling 712100China

This study was conducted to investigate potential regulatory mechanisms of feed efficiency(FE)in sheep by linking rumen microbiota with its host by the multi-omics *** hundred and ninety-eight hybrid female sheep(initial body weight=30.88±4.57 kg;4-month-old)were selected as candidate *** test sheep was fed in an individual pen for 60 days,and the residual feed intake(RFI)was *** ten candidate sheep with the highest RFI were divided into the Low-FE group,and the ten with the lowest RFI were divided into the High-FE group,all selected for sample *** RFI,average daily gain and average daily feed intake were highly significantly different between the two experimental groups(Pkey bacteria,and increased abundance of the genes encoding the enzymes for cellulose degradation and production of propionate in High-FE *** results of proteomics and section showed the rumen papilla length and expression of carbonic anhydrase and Na^(+)/K^(+)-ATPase were significantly higher in High-FE group(P<0.05).On the other hand,the acetyl-CoA content significantly increased in the liver of High-FE group(P=0.002).The relative expression levels of insulin-like growth factor-1 and apolipoprotein A4 genes were significantly up-regulated in the liver of High-FE group(P<0.05),but relative expression level of monoacylglycerol O-acyltransferase 3 gene was significantly down-regulated(P=0.037).These findings provide the mechanism by which the collaborative interaction between rumen microbiota fermentation and host uptake and metabolism of fermentation products impacts feed efficiency tr

关键词： Residual feed intake Sheep Microbial composition Rumen proteome Liver transcriptome Metabolite

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miR-24-3p promotes proliferation and inhibits apoptosis of porcine granulosa cells by targeting P27

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Journal of Integrative Agriculture 2024年第4期23卷 1315-1328页

作者： Shengjie Shi Lutong Zhang Liguang Wang Huan Yuan Haowei Sun Mielie Madaniyati Chuanjiang Cai Weijun Pang Lei Gao Guiyan Chu College of animal Science and Technology Northwest A&F UniversityYangling 712100China key laboratory of animal genetics Breeding and Reproduction of Shaanxi ProvinceYangling 712100China

Ovarian follicle development is associated with the physiological functions of granulosa cells(GCs),including proliferation and *** level of miR-24-3p in ovarian tissue of high-yielding Yorkshire×Landrace sows was significantly higher than that of low-yielding ***,the functions of miR-24-3p on GCs are *** this study,using flow cytometry,5-ethynyl-2′-de-oxyuridine(EdU)staining,and cell count,we showed that miR-24-3p promoted the proliferation of GCs increasing the proportion of cells in the S phase and upregulating the expression of cell cycle genes,moreover,miR-24-3p inhibited GC ***,on-line prediction,bioinformatics analysis,a luciferase reporter assay,RT-qPCR,and Western blot results showed that the target gene of miR-24-3p in proliferation and apoptosis is cyclin-dependent kinase inhibitor 1B(P27/CDKN1B).Furthermore,the effect of miR-24-3p on GC proliferation and apoptosis was attenuated by P27 *** findings suggest that miR-24-3p regulates the physiological functions of GCs.

关键词： miR-24-3p granulosa cells proliferation apoptosis

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MicroRNA-382 inhibits the proliferation of mouse spermatogoniaby targeting Kmt5a

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BIOCELL 2020年第2期44卷 201-207页

作者： YI ZHENG PENGFEI ZHANG JINSHENG ZHU LINGKAI ZHANG WENXIAN ZENG key laboratory for animal genetics Breeding and Reproduction of Shaanxi ProvinceCollege of Animal Science and TechnologyNorthwest A&F UniversityYangling712100China

Spermatogenesis is a highly efficient and intricate process in the testis by which mature spermatozoa are produced daily to maintain lifelong male *** to this process are spermatogonia capable of both proliferation and ***,the underlying mechanisms for spermatogonial proliferation and differentiation remain poorly ***(miRNAs)are a category of non-coding small RNAs with regulatory functions by binding to the 3’untranslated region(UTR)of the target *** studies have demonstrated that miRNAs are capable of modulating cell proliferation,differentiation and apoptosis,but the roles of individual miRNAs in spermatogonial fate determination remain largely ***,by using a mouse spermatogonial cell line(GC-1),we investigated the role for miRNA-382 in spermatogonial *** found that pre-miRNA-382 was expressed in *** luciferase reporter assay demonstrated Kmt5a but not Top1 as a target gene of *** of miRNA-382 by transfecting a miRNA mimic downregulated Kmt5a at both RNA and protein levels,and further reduced the proliferation and viability of *** of Kmt5a by RNA interference(RNAi)resulted in a uniform phenotype in *** therefore conclude that miRNA-382 inhibits the proliferation of mouse spermatogonia by targeting *** finding extends the knowledge about the regulatory roles of miRNAs in spermatogonia and lays the groundwork for diagnosis and treatment of male infertility.

关键词： Spermatogenesis Self-renewal Testis

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