检索结果-南通市图书馆

Proteomics of spermatogenesis： from protein lists to understanding the regulation of male fertility and infertility

在线全文

学校读者我要写书评

暂无评论

Asian Journal of Andrology 2011年第1期13卷 18-23页

作者： Xiaoyan Huang Jiahao Sha Lab of Reproductive Medicine Department of Histology and Embryology Nanjing Medical University Nanjing 210029 China

Proteomic technologies have undergone significant development in recent years, which has led to extensive advances in protein research. Currently, proteomic approaches have been applied to many scientific areas, including basic research, various disease and malignant tumour diagnostics, biomarker discovery and other therapeutic applications. In addition, proteomics-driven research articles examining reproductive biology and medicine are becoming increasingly common. The key challenge for this field is to move from lists of identified proteins to obtaining biological information regarding protein function. The present article reviews the available scientific literature related to spermatogenesis. In addition, this study uses two-dimensional electrophoresis mass spectrometry （2DE-MS） and liquid chromatography （LC）-MS to construct a series of proteome profiles describing spermatogenesis. This large-scale identification of proteins provides a rich resource for elucidating the mechanisms underlying male fertility and infertility.

关键词： infertility male fertility proteomics spermatogenesis

The expression and localization of a novel protein phosphatase inhibitor 2810408A11Rik in mouse testis and sperm

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

The Journal of Biomedical Research 2012年第2期26卷 110-116页

作者： Ye Bi Mingxi Liu Wenjiao Tu Yibo Wu Xuejiang Guo Zuomin Zhou Jiahao Sha State Key Laboratory of Reproductive Medicine Department of Histology and EmbryologyNanjing Medical UniversityNanjingJiangsu 210029China

This study investigated the expression and distribution of 2810408A11Rik in mouse testis and sperm, and explored its role in sperrnatogenesis and sperm function. The expression levels of 2810408A11Rik mRNA in multiple tissue samples were analyzed using bioinformatic resources and RT-PCR technique. A specific rabbit polyclonal antibody was prepared by prokaryotic expression of 2810408A11Rik recombinant protein and utilized for animal immunization. Western blotting, immunohistochemistry and immunofluorescence were used to detect the expression and distribution of 2810408A11Rik. The results of the bioinformatic analysi and RT-PCR showed that 2810408A11Rik mRNA was specifically expressed in mouse testis, and 2810408AllRik protein included a protein phosphatase inhibitor domain. Western blotting assays, immunohistochemistry and immunofluorescence confirmed the expression of 2810408A11Rik protein in mouse testis, especially in post-meiosis round and long spermatids, and that it is localized in the acrosome and the post-nucleus area of sperm. Our findings suggest that 2810408A11Rik may play an important role in spermatogenesis, sperm capacitation and fertilization.

关键词： 2810408A11Rik testis specific protein phosphatase inhibitor capacitation

RNA Guided Genome Editing in Mouse Germ-Line Stem Cells

在线全文

学校读者我要写书评

暂无评论

Journal of Genetics and Genomics 2014年第7期41卷 409-411页

作者： Xuepeng Wang Yan Yuan Quan Zhou Haifeng Wan Mei Wang Qi Zhou Xiao-Yang Zhao Jiahao Sha State Key Laboratory of Reproductive Biology Institute of ZoologyChinese Academy of Sciences University of Chinese Academy of Sciences State Key Laboratory of Reproductive Medicine Institute of ToxicologyNanjing Medical University

Spermatogonial stem cells （SSCs） reside on the basement membrane of the seminiferous tubules in mammalian testes （Nagano et al., 1998）. After isolation and purification of SSCs from mouse testis, SSCs can be cultured in vitro to derive germ-line stem cells （GSCs） which have the ability of proliferation over 2 years （Kanatsu-Shinohara et al.. 2003;

关键词： EGFP In

dRSPH3BP recruited dRSPH3 building Radical spoke during spermatogenesis for activity and phosphorylation in axoneme

在线全文

学校读者我要写书评

暂无评论

dRSPH3BP recruited dRSPH3 building Radical spoke during sper...

A susceptibility locus rs7099208 is associated with non-obstructive azoospermia via reduction in the expression of FAM160B1

中国生理学会生殖科学专业委员会—中国动物学会生殖生物学...

作者： Ya Wang Juan Huang MingXi Liu Jiahao Sha State Key Laboratory of Reproductive Medicine Nanjing Medical University

＜正＞Primary ciliary dyskinesia（PCD）affects one in 15,000-30,000 individuals,composed of a heterogeneous group of autosomal-recessive disorders.We used UAS GAL4 system to screen genes involved in PCD,especially in spermatogenesis and identified these genes on which made knockout（KO）in Drosophila by Cas9/CRISPR system.Of all genes KO lines,dRSPH3BP KO flies are male sterile.

关键词：

来源： cnki会议评论

在线全文

cnki会议

学校读者我要写书评

暂无评论

The Journal of Biomedical Research 2015年第6期29卷 491-500页

作者： Yan Zhang Jing Qian Minghui Wu Mingxi Liu Kai Zhang Yuan Lin Xuejiang Guo Zuomin Zhou Zhibin Hu Jiahao Sha State Key Laboratory of Reproductive Medicine Nanjing Medical University The Center for Reproductive Medicine The Second Affiliated Hospital of Nanjing Medical University Department of Histology and Embryology Nanjing Medical University Department of Epidemiology and Biostatistics and Key Laboratory of Modern Toxicology of Ministry of Education Nanjing Medical University

Non-obstructive azoospermia （NOA） is a severe defect in male reproductive health that occurs in 1% of adult men. In a previous study, we identified that rs7099208 is located within the last intron of FAM160B1 at 10q25.3. In this study, we analysed expression Quantitative Trait Loci （eQTL） of FAM16OB1, ABLIM1 and TRUB1, the three genes surrounding rs7099208. Only the expression level of FAM16OB1 was reduced for the homozygous alternate genotype （GG） of rs7099208, but not for the homozygous reference or heterozygous geno- types. FAM160B1 is predominantly expressed in human testes, where it is found in spermatocytes and round sper- matids. From 17 patients with NOA and five with obstructive azoospermia （OA）, immunohistochemistry revealed that expression of FAM160B1 is reduced, or undetectable in NOA patients, but not in OA cases or normal men. We conclude that rs7099208 is associated with NOA via a reduction in the expression of FAM160B1.

关键词： non-obstructive azoospermia obstructive azoospermia rs7099208 FAM160B1 expressionQuantitative Trait Loci apoptosis

CircAST:Full-length Assembly and Quantification of Alternatively Spliced Isoforms in Circular RNAs

在线全文

学校读者我要写书评

暂无评论

Genomics, Proteomics & Bioinformatics 2019年第5期17卷 522-534页

作者： Jing Wu Yan Li Cheng Wang Yiqiang Cui Tianyi Xu Chang Wang Xiao Wang Jiahao Sha Bin Jiang Kai Wang Zhibin Hu Xuejiang Guo Xiaofeng Song Department of Biomedical Engineering Nanjing University of Aeronautics and AstronauticsNanjing 211106China School of Biomedical Engineering and Informatics Nanjing Medical UniversityNanjing 211166China State Key Laboratory of Reproductive Medicine Nanjing Medical UniversityNanjing 211166China Center of Pathology and Clinical Laboratory Sir Run Run HospitalNanjing Medical UniversityNanjing 211166China Center for Cellular and Molecular Therapeutic Children’s Hospital of PhiladelphiaPhiladelphiaPA 19104USA

Circular RNAs(circ RNAs),covalently closed continuous RNA loops,are generated from cognate linear RNAs through back splicing events,and alternative splicing events may generate different circ RNA isoforms at the same locus.However,the challenges of reconstruction and quantification of alternatively spliced full-length circ RNAs remain unresolved.On the basis of the internal structural characteristics of circ RNAs,we developed Circ AST,a tool to assemble alternatively spliced circ RNA transcripts and estimate their expression by using multiple splice graphs.Simulation studies showed that Circ AST correctly assembled the full sequences of circ RNAs with a sensitivity of 85.63%–94.32%and a precision of 81.96%–87.55%.By assigning reads to specific isoforms,Circ AST quantified the expression of circ RNA isoforms with correlation coefficients of 0.85–0.99 between theoretical and estimated values.We evaluated Circ AST on an in-house mouse testis RNA-seq dataset with RNase R treatment for enriching circ RNAs and identified 380 circ RNAs with full-length sequences different from those of their corresponding cognate linear RNAs.RT-PCR and Sanger sequencing analyses validated 32 out of 37 randomly selected isoforms,thus further indicating the good performance of Circ AST,especially for isoforms with low abundance.We also applied Circ AST to published experimental data and observed substantial diversity in circular transcripts across samples,thus suggesting that circ RNA expression is highly regulated.Circ AST can be accessed freely at https://***/xiaofengsong/CircAST.

关键词： Circular RNA Full-length reconstruction Isoform quantification Multiple splice graph model Transcriptome

RSBP15 interacts with and stabilizes dRSPH3 during sperm axoneme assembly in Drosophila

在线全文

学校读者我要写书评

暂无评论

Journal of Genetics and Genomics 2019年第6期46卷 281-290页

作者： Ya Wang Rui Xu Yiwei Cheng Haowei Cao Zibin Wang Tianyu Zhu Jiayin Jiang Hao Zhang Chang Wang Lin Qi Mingxi Liu Xuejiang Guo Juan Huang Jiahao Sha State Key Laboratory of Reproductive Medicine Nanjing Medical University Nanjing 211166 China Department of Histology and Embryology Nanjing Medical University Nanjing 211166 China Department of Medical Genetics Nanjing Medical University Nanjing 211166 China Jiangsu Key Laboratory of Xenotransplantation Nanjing Medical University Nanjing 211166 China

Flagellum in sperm is composed of over 200 different proteins and is essential for sperm motility. In particular, defects in the assembly of the radial spoke in the flagellum result in male infertility due to loss of sperm motility. However, mechanisms regulating radial spoke assembly remain unclear in metazoans.Here, we identified a novel Drosophila protein radial spoke binding protein 15(RSBP15) which plays an important role in regulating radial spoke assembly. Loss of RSBP15 results in complete lack of mature sperms in seminal vesicles(SVs), asynchronous individualization complex(IC) and defective "9 + 2"structure in flagella. RSBP15 is colocalized with dRSPH3 in sperm flagella, and interacts with dRSPH3 through its DD_R_PKA superfamily domain which is important for the stabilization of dRSPH3. Moreover,loss of dRSPH3, as well as dRSPH1, dRSPH4 a and dRSPH9, showed similar phenotypes to rsbp15 KO mutant. Together, our results suggest that RSBP15 acts in stabilizing the radial spoke protein complex to anchor and strengthen the radial spoke structures in sperm flagella.

关键词： RSBP15 RSPH3 Flagellum Radial spoke Drosophila

Flotillin-2 is an acrosome-related protein involved in mouse spermiogenesis

在线全文

学校读者我要写书评

暂无评论

The Journal of Biomedical Research 2012年第4期26卷 278-287页

作者： Yibo Wu Xin Chen Shuai Wang Min Jiang Bo Zheng Quan Zhou Ye Bi Zuomin Zhou Xiaoyan Huang Jiahao Sha State Key Laboratory of Reproductive Medicine Department of Histology and EmbryologyNanjing Medical UniversityNanjingJiangsu 210029China

Spermatogenesis is a complex process of terminal differentiation by which mature sperms are generated,and it can be divided into three phases：mitosis,meiosis and spermiogenesis.In a previous study,we established a series of proteomic profiles for spermatogenesis to understand the regulation of male fertility and infertility.Here,we further investigated the localization and the role of flotillin-2 in spermiogenesis.Flotillin-2 expression was investigated in the testis of male CD1 mice at various developmental stages of spermatogenesis by using Western blotting,immunohistochemistry and immunofluorescence.Flotillin-2 was knocked down in vivo in three-week-old male mice using intratesticular injection of small inhibitory RNA（siRNA）,and sperm abnormalities were assessed three weeks later.Flotillin-2 was expressed at high levels in male germ cells during spermatogenesis.Flotillin-2 immunoreactivity was observed in pachytene spermatocytes as a strong dot-shaped signal and in round spermatids as a sickle-shaped distribution ahead of the acrosome.Immunofluorescence confirmed flotillin-2 was localized in front of the acrosome in round spermatids,indicating that flotillin-2 was localized to the Golgi apparatus.Knockdown of flotillin-2 in vivo led to a significant increase in head sperm abnormalities isolated from the cauda epididymis,compared with control siRNA-injected testes.This study indicates that flotillin-2 is a novel Golgi-related protein involved in sperm acrosome biogenesis.

关键词： flotillin-2 Golgi apparatus RNA interference acrosome biogenesis

DDX3X regulates cell survival and cell cycle during mouse early embryonic development

在线全文

学校读者我要写书评

暂无评论

The Journal of Biomedical Research 2014年第4期28卷 282-291页

作者： Qian Li Pan Zhang Chao Zhang Ying Wang Ru Wan Ye Yang Xuejiang Guo Ran Huo Min Lin Zuomin Zhou Jiahao Sha State Key Laboratory of Reproductive Medicine Nanjing Medical University

DDX3X is a highly conserved DEAD-box RNA helicase that participates in RNA transcription, RNA splicing, and mRNA transport, translation, and nucleo-cytoplasmic transport. It is highly expressed in metaphase II （MII） oocytes and is the predominant DDX3 variant in the ovary and embryo. However, whether it is important in mouse early embryo development remains unknown. In this study, we investigated the function of DDX3X in early embryogenesis by cytoplasmic microinjection with its siRNA in zygotes or single blastomeres of 2-cell embryos. Our results showed that knockdown of Ddx3x in zygote cytoplasm led to dramatically diminished blastocyst formarion, reduced cell numbers, and an increase in the number of apoptotic cells in blastocysts. Meanwhile, there was an accumulation of p53 in RNAi blastocysts. In addition, the ratio of cell cycle arrest during 2-cell to 4-cell transition increased following microinjection of Ddx3x siRNA into single blastomeres of 2-cell embryos compared with control. These results suggest that Ddx3x is an essential gene associated with cell survival and cell cycle control in mouse early embryos, and thus plays key roles in normal embryo development.

关键词： DDX3X early embryo p53 apoptosis cell cycle