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Min Yu Obstetrics & Gynecology Hospital of Fudan University, Shanghai JIAI Genetics & IVF Institute, Shanghai, China

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Xiandong Peng Obstetrics & Gynecology Hospital of Fudan University, Shanghai JIAI Genetics & IVF Institute, Shanghai, China

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He Li Obstetrics & Gynecology Hospital of Fudan University, Shanghai JIAI Genetics & IVF Institute, Shanghai, China

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Yining Xu Obstetrics & Gynecology Hospital of Fudan University, Shanghai JIAI Genetics & IVF Institute, Shanghai, China

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Xiaoxi Sun Obstetrics & Gynecology Hospital of Fudan University, Shanghai JIAI Genetics & IVF Institute, Shanghai, China
Shanghai Key Laboratory of Female Reproductive and Endocrine-Related Diseases, Shanghai, China

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Jiazhou Chen Obstetrics & Gynecology Hospital of Fudan University, Shanghai JIAI Genetics & IVF Institute, Shanghai, China

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Embryo implantation, a critical step during the mammalian reproductive process, requires normal developing blastocysts and a receptive endometrium. Endometriosis, a common pathologically benign gynecological condition, is associated with decreased fertility and reduced endometrial receptivity. The oncoprotein, Gankyrin, has been associated with endometriosis and endometrial cancer. Here, we examined the role of Gankyrin during the process of embryo implantation and found that Gankyrin expression levels were significantly increased during the mid-secretory phase, but unaffected during the proliferative phase in the human endometrium. Using an in vitro cell adhesion assay to examine the cell adhesion rate of BeWo trophoblast spheroids to Gankyrin knockdown or overexpressing human endometrial carcinoma RL95-2 cells, we demonstrated that the adhesion rate was significantly reduced in Gankyrin-knockdown RL95-2 cells, while overexpression of Gankyrin promoted cell adhesion. Furthermore, we found that the downregulation of Gankyrin inhibited STAT3 activation and subsequent matrix metalloproteinase 2 (MMP2) expression, while overexpression led to STAT3 activation and MMP2 expression. In vivo, we found that Gankyrin expression was increased in the endometrium after conception but decreased with the prolongation of gestation time in female mice. siRNA-mediated knockdown of Gankyrin in the uterine horn led to a significant reduction in the number of implanted embryos 9 days post-gestation, which was associated with a decrease in p-STAT3 expression and MMP2 transcription. Taken together, our findings indicate that Gankryin has a potential role in embryo implantation via STAT3 activation.

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Elizabeth M Parrish Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA

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Anaar Siletz Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA
Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA
Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA

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Min Xu Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA
Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA
Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA

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Teresa K Woodruff Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA
Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA
Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA
Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA
Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA

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Lonnie D Shea Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA
Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA
Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA
Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA
Department of Chemical and Biological Engineering, Robert H Lurie Comprehensive Cancer Center, Chemistry of Life Processes Institute, Institute for BioNanotechnology in Advanced Medicine, Department of Obstetrics and Gynecology, Department of Biochemistry, Member of the Oncofertility Consortium, Medical Scientist Training Program, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, Illinois 60208, USA

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Ovarian follicle maturation results from a complex interplay of endocrine, paracrine, and direct cell–cell interactions. This study compared the dynamic expression of key developmental genes during folliculogenesis in vivo and during in vitro culture in a 3D alginate hydrogel system. Candidate gene expression profiles were measured within mouse two-layered secondary follicles, multi-layered secondary follicles, and cumulus–oocyte complexes (COCs). The expression of 20 genes involved in endocrine communication, growth signaling, and oocyte development was investigated by real-time PCR. Gene product levels were compared between i) follicles of similar stage and ii) COCs derived either in vivo or by in vitro culture. For follicles cultured for 4 days, the expression pattern and the expression level of 12 genes were the same in vivo and in vitro. Some endocrine (cytochrome P450, family 19, subfamily A, polypeptide 1 (Cyp19a1) and inhibin βA subunit (Inhba)) and growth-related genes (bone morphogenetic protein 15 (Bmp15), kit ligand (Kitl), and transforming growth factor β receptor 2 (Tgfbr2)) were downregulated relative to in vivo follicles. For COCs obtained from cultured follicles, endocrine-related genes (inhibin α-subunit (Inha) and Inhba) had increased expression relative to in vivo counterparts, whereas growth-related genes (Bmp15, growth differentiation factor 9, and kit oncogene (Kit)) and zona pellucida genes were decreased. However, most of the oocyte-specific genes (e.g. factor in the germline α (Figla), jagged 1 (Jag1), and Nlrp5 (Mater)) were expressed in vitro at the same level and with the same pattern as in vivo-derived follicles. These studies establish the similarities and differences between in vivo and in vitro cultured follicles, guiding the creation of environments that maximize follicle development and oocyte quality.

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Hui Wang School of Food Science, Department of Animal Science, Department of Animal Sciences, School of Food Science, Washington State University, Pullman, Washington 99164, USA
School of Food Science, Department of Animal Science, Department of Animal Sciences, School of Food Science, Washington State University, Pullman, Washington 99164, USA

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Yansong Xue School of Food Science, Department of Animal Science, Department of Animal Sciences, School of Food Science, Washington State University, Pullman, Washington 99164, USA

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Baolin Wang School of Food Science, Department of Animal Science, Department of Animal Sciences, School of Food Science, Washington State University, Pullman, Washington 99164, USA

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Junxing Zhao School of Food Science, Department of Animal Science, Department of Animal Sciences, School of Food Science, Washington State University, Pullman, Washington 99164, USA

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Xu Yan School of Food Science, Department of Animal Science, Department of Animal Sciences, School of Food Science, Washington State University, Pullman, Washington 99164, USA

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Yan Huang School of Food Science, Department of Animal Science, Department of Animal Sciences, School of Food Science, Washington State University, Pullman, Washington 99164, USA

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Min Du School of Food Science, Department of Animal Science, Department of Animal Sciences, School of Food Science, Washington State University, Pullman, Washington 99164, USA

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Mei-Jun Zhu School of Food Science, Department of Animal Science, Department of Animal Sciences, School of Food Science, Washington State University, Pullman, Washington 99164, USA
School of Food Science, Department of Animal Science, Department of Animal Sciences, School of Food Science, Washington State University, Pullman, Washington 99164, USA

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Accompanying the dramatic increase in maternal obesity, the incidence of type 1 diabetes (T1D) in children is also rapidly increasing. The objective of this study was to explore the effects of maternal obesity on the incidence of T1D in offspring using non-obese diabetic (NOD) mice, a common model for TID. Four-week-old female NOD mice were fed either a control diet (10% energy from fat, CON) or a high-fat diet (60% energy from fat) for 8 weeks before mating. Mice were maintained in their respective diets during pregnancy and lactation. All offspring mice were fed the CON to 16 weeks. Female offspring (16-week-old) born to obese dams showed more severe islet lymphocyte infiltration (major manifestation of insulitis) (P<0.01), concomitant with elevated nuclear factor kappa-light-chain-enhancer of activated B cells p65 signaling (P<0.01) and tumor necrosis factor alpha protein level (P<0.05) in the pancreas. In addition, maternal obesity resulted in impaired (P<0.05) glucose tolerance and lower (P<0.05) serum insulin levels in offspring. In conclusion, maternal obesity resulted in exacerbated insulitis and inflammation in the pancreas of NOD offspring mice, providing a possible explanation for the increased incidence of T1D in children.

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Hadrian M Kinnear Program in Cellular and Molecular Biology, University of Michigan, Ann Arbor, Michigan, USA
Medical Scientist Training Program, University of Michigan, Ann Arbor, Michigan, USA

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Claire E Tomaszewski Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan, USA

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Faith L Chang Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan, USA

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Molly B Moravek Department of Obstetrics and Gynecology, University of Michigan, Ann Arbor, Michigan, USA
Division of Reproductive Endocrinology and Infertility, University of Michigan, Ann Arbor, Michigan, USA
Department of Urology, University of Michigan, Ann Arbor, Michigan, USA

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Min Xu Department of Obstetrics and Gynecology, University of Michigan, Ann Arbor, Michigan, USA
Division of Reproductive Endocrinology and Infertility, University of Michigan, Ann Arbor, Michigan, USA

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Vasantha Padmanabhan Department of Obstetrics and Gynecology, University of Michigan, Ann Arbor, Michigan, USA
Department of Pediatrics and Communicable Diseases, University of Michigan, Ann Arbor, Michigan, USA

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Ariella Shikanov Program in Cellular and Molecular Biology, University of Michigan, Ann Arbor, Michigan, USA
Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan, USA
Department of Obstetrics and Gynecology, University of Michigan, Ann Arbor, Michigan, USA

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Historically, research in ovarian biology has focused on folliculogenesis, but recently the ovarian stroma has become an exciting new frontier for research, holding critical keys to understanding complex ovarian dynamics. Ovarian follicles, which are the functional units of the ovary, comprise the ovarian parenchyma, while the ovarian stroma thus refers to the inverse or the components of the ovary that are not ovarian follicles. The ovarian stroma includes more general components such as immune cells, blood vessels, nerves, and lymphatic vessels, as well as ovary-specific components including ovarian surface epithelium, tunica albuginea, intraovarian rete ovarii, hilar cells, stem cells, and a majority of incompletely characterized stromal cells including the fibroblast-like, spindle-shaped, and interstitial cells. The stroma also includes ovarian extracellular matrix components. This review combines foundational and emerging scholarship regarding the structures and roles of the different components of the ovarian stroma in normal physiology. This is followed by a discussion of key areas for further research regarding the ovarian stroma, including elucidating theca cell origins, understanding stromal cell hormone production and responsiveness, investigating pathological conditions such as polycystic ovary syndrome (PCOS), developing artificial ovary technology, and using technological advances to further delineate the multiple stromal cell types.

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Xiaoyan Huang Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, 210029, P.R. China and Division of Pharmacology Research, China Pharmaceutical University, Nanjing, 210009, P.R. China

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Jun Zhang Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, 210029, P.R. China and Division of Pharmacology Research, China Pharmaceutical University, Nanjing, 210009, P.R. China

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Li Lu Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, 210029, P.R. China and Division of Pharmacology Research, China Pharmaceutical University, Nanjing, 210009, P.R. China

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Lanlan Yin Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, 210029, P.R. China and Division of Pharmacology Research, China Pharmaceutical University, Nanjing, 210009, P.R. China

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Min Xu Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, 210029, P.R. China and Division of Pharmacology Research, China Pharmaceutical University, Nanjing, 210009, P.R. China

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Youqun Wang Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, 210029, P.R. China and Division of Pharmacology Research, China Pharmaceutical University, Nanjing, 210009, P.R. China

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Zuomin Zhou Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, 210029, P.R. China and Division of Pharmacology Research, China Pharmaceutical University, Nanjing, 210009, P.R. China

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Jiahao Sha Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, 210029, P.R. China and Division of Pharmacology Research, China Pharmaceutical University, Nanjing, 210009, P.R. China

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Identification of genes specifically expressed in adult and fetal testis is important in furthering our understanding of testis development and function. In this study, a novel human transcript, designated human testis cAMP-responsive element-binding protein (htCREB), was identified by hybridization of adult and fetal human testis cDNA probes with a human cDNA microarray containing 9216 clones. The htCREB transcript (GenBank Accession no. AY347527) was expressed at 2.35-fold higher levels in adult human testes than in fetal testes. Sequence and ntBLAST analyses against the human genome database indicated that htCREB was a novel splice variant of human CREB. RT-PCR-based tissue distribution experiments demonstrated that the htCREB transcript was highly expressed in adult human testis and in healthy sperm, but not in testes from patients with Sertoli cell-only syndrome. Taken together, these results suggest that the htCREB transcript is chiefly expressed in germ cells and is most likely involved in spermatogenesis.

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Jingbo Dai School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China

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Wangjie Xu School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China

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Xianglong Zhao School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China

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Meixing Zhang School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China

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Dong Zhang School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China

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Dongsheng Nie School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China

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Min Bao School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China

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Zhaoxia Wang School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China

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Lianyun Wang School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China

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Zhongdong Qiao School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China

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Many studies have revealed the hazardous effects of cigarette smoking and nicotine exposure on male fertility, but the actual, underlying molecular mechanism remains relatively unclear. To evaluate the detrimental effects of nicotine exposure on the sperm maturation process, two-dimensional gel electrophoresis and mass spectrometry analyses were performed to screen and identify differentially expressed proteins from the epididymal tissue of mice exposed to nicotine. Data mining analysis indicated that 15 identified proteins were mainly involved in the molecular transportation process and the polyol pathway, indicating impaired epididymal secretory functions. Experiments in vitro confirmed that nicotine inhibited tyrosine phosphorylation levels in capacitated spermatozoa via the downregulated seminal fructose concentration. Sord, a key gene encoding sorbitol dehydrogenase, was further investigated to reveal that nicotine induced hyper-methylation of the promoter region of this gene. Nicotine-induced reduced expression of Sord could be involved in impaired secretory functions of the epididymis and thus prevent the sperm from undergoing proper maturation and capacitation, although further experiments are needed to confirm this hypothesis.

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Shi-Yu An Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, China
State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, China

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Zi-Fei Liu Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, China

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El-Samahy M A Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, China

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Ming-Tian Deng Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, China

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Xiao-Xiao Gao Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, China

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Ya-Xu Liang Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, China

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Chen-Bo Shi Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, China

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Zhi-Hai Lei College of veterinary medicine, Nanjing Agricultural University, Nanjing, China

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Feng Wang Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, China

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Guo-Min Zhang Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing, China
College of veterinary medicine, Nanjing Agricultural University, Nanjing, China

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Long ncRNAs regulate a complex array of fundamental biological processes, while its molecular regulatory mechanism in Leydig cells (LCs) remains unclear. In the present study, we established the lncRNA LOC102176306/miR-1197-3p/peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PPARGC1A) regulatory network by bioinformatic prediction, and investigated its roles in goat LCs. We found that lncRNA LOC102176306 could efficiently bind to miR-1197-3p and regulate PPARGC1A expression in goat LCs. Downregulation of lncRNA LOC102176306 significantly supressed testosterone (T) synthesis and ATP production, decreased the activities of antioxidant enzymes and mitochondrial complex I and complex III, caused the loss of mitochondrial membrane potential, and inhibited the proliferation of goat LCs by decreasing PPARGC1A expression, while these effects could be restored by miR-1197-3p inhibitor treatment. In addition, miR-1197-3p mimics treatment significantly alleviated the positive effects of lncRNA LOC102176306 overexpression on T and ATP production, antioxidant capacity and proliferation of goat LCs. Taken together, lncRNA LOC102176306 functioned as a sponge for miR-1197-3p to maintain PPARGC1A expression, thereby affecting the steroidogenesis, cell proliferation and oxidative stress of goat LCs. These findings extend our understanding of the molecular mechanisms of T synthesis, cell proliferation and oxidative stress of LCs.

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