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Jiarong Feng Department of Infertility and Sexual Medicine, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou, Guangdong, China

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Yanan Zhang Department of Infertility and Sexual Medicine, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou, Guangdong, China

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Xiaojian Yang Department of Infertility and Sexual Medicine, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou, Guangdong, China

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Yan Zhang Department of Infertility and Sexual Medicine, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou, Guangdong, China

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In brief

The genetic heterogeneity of CFTR gene mutations in Chinese patients with congenital absence of the vas deferens (CAVD) differs from the hotspot mutation pattern in Caucasians. This paper reviews and suggests a more suitable screening strategy for the Chinese considering the dilemma of CFTR genetic blocking.

Abstract

Congenital absence of the vas deferens (CAVD) is a major cause of obstructive azoospermia and male infertility, with CFTR gene mutation as the main pathogenesis. Other genes such as ADGRG2, SLC9A3, and PANK2 have been discovered and proven to be associated with CAVD in recent studies. Multiple CFTR hotspot mutations have been found in Caucasians in several foreign countries, and relevant genetic counseling and preimplantation genetic diagnosis (PGD) have been conducted for decades. However, when we examined research on Chinese CAVD, we discovered that CFTR mutations show heterogeneity in the Chinese Han population, and there is currently no well-established screening strategy. Therefore, we have reviewed the literature, combining domestic and international research as well as our own, aiming to review research progress on the CFTR gene in China and discuss the appropriate scope for CFTR gene detection, the detection efficiency of other CAVD-related genes, and the screening strategy applicable to the Chinese Han population. This study provides more valuable information for genetic counseling and a theoretical basis for PGD and treatment for couples with CAVD when seeking reproductive assistance.

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Haolin Zhang Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China
Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China
Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China

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Ming Yi Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China

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Yan Zhang Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China
Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China

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Hongyan Jin Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China
Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China

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Wenxin Zhang Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China
Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China

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Jingjing Yang Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China
Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China

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Liying Yan Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China
Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China

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Rong Li Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China
Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China

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Yue Zhao Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China
Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China

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Jie Qiao Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China
Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China
Department of Obstetrics and Gynaecology, Key Laboratory of Assisted Reproduction, Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Neuroscience Research Institute, Center for Reproductive Medicine, Peking University Third Hospital, No. 49, North Huayuan Road, Haidian District, Beijing 100191, China

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Polycystic ovary syndrome (PCOS) is a complex endocrine and metabolic disorder with unclear etiology and unsatisfactory management. Effects of diets on the phenotype of PCOS were not fully understood. In the present study, we applied 45 and 60% high-fat diets (HFDs) on a rat model of PCOS induced by postnatal DHEA injection. We found that both DHEA and DHEA+HFDs rats exhibited reproductive abnormalities, including hyperandrogenism, irregular cycles and polycystic ovaries. The addition of HFDs, especially 60% HFDs, exaggerated morphological changes of ovaries and a number of metabolic changes, including increased body weight and body fat content, impaired glucose tolerance and increased serum insulin levels. Results from qPCR showed that DHEA-induced increased expression of hypothalamic androgen receptor and LH receptor were reversed by the addition of 60% HFDs. In contrast, the ovarian expression of LH receptor and insulin receptor mRNA was upregulated only with the addition of 60% HFDs. These findings indicated that DHEA and DHEA+HFDs might influence PCOS phenotypes through distinct mechanisms: DHEA affects the normal function of hypothalamus–pituitary–ovarian axis through LH, whereas the addition of HFDs exaggerated endocrine and metabolic dysfunction through ovarian responses to insulin-related mechanisms. We concluded that the addition of HFDs yielded distinct phenotypes of DHEA-induced PCOS and could be used for studies on both reproductive and metabolic features of the syndrome.

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Nan Meng NHC Key Lab of Reproduction Regulation, Shanghai Institute of Planned Parenthood Research, Shanghai, People’s Republic of China

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Xinyue Wang Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, People’s Republic of China

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Yan Shi NHC Key Lab of Reproduction Regulation, Shanghai Institute of Planned Parenthood Research, Shanghai, People’s Republic of China

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Yanyan Mao NHC Key Lab of Reproduction Regulation, Shanghai Institute of Planned Parenthood Research, Shanghai, People’s Republic of China

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Qian Yang NHC Key Lab of Reproduction Regulation, Shanghai Institute of Planned Parenthood Research, Shanghai, People’s Republic of China

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Baohui Ju Department of Gynecology and Obstetrics, The Second Hospital of Tianjin Medical University, Tianjin, People’s Republic of China

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Qianxi Zhu NHC Key Lab of Reproduction Regulation, Shanghai Institute of Planned Parenthood Research, Shanghai, People’s Republic of China

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Tingting Zhang Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, People’s Republic of China

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Yan Gu Department of Gynecology and Obstetrics, The Second Hospital of Tianjin Medical University, Tianjin, People’s Republic of China

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Xuan Zhang NHC Key Lab of Reproduction Regulation, Shanghai Institute of Planned Parenthood Research, Shanghai, People’s Republic of China

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Decidualization is essential for the successful establishment of pregnancy, and the dysregulated decidualization may lead to early pregnancy loss. It was previously reported by us that miR-3074-5p could promote apoptosis but inhibit invasion of human extravillous trophoblast (EVT) cells in vitro, and the expression level of miR-3074-5p in villus tissues of recurrent miscarriage (RM) patients was significantly increased. The aim of this study was to preliminarily explore the role of miR-3074-5p played in the decidualization of human endometrial stromal cells (ESCs). It was found that the decidual expression level of miR-3074-5p in RM patients was remarkably higher than that in the control group. The overexpression of miR-3074-5p in the immortalized human ESC line, T-HESCs, showed suppressive effects not only on the cell proliferation, as well as the intracellular expression levels of cyclin B1 (CCNB1), CCND1 and CCNE1 but also on the in vitro-induced decidualization. CLN8 mRNA, encoding an endoplasmic reticulum (ER)-associated membrane protein, was validated to be directly targeted by miR-3074-5p. And, the expression level of CLN8 was continuously increased along with the decidualization process, whereas down-regulated CLN8 expression could inhibit the decidualization of T-HESCs in vitro. Furthermore, contrary to the increased expression level of miR-3074-5p, a significantly decreased CLN8 expression was observed in decidual tissues of RM patients. Collectively, these data suggested that an increased miR-3074-5p expression in ESCs might cause early pregnancy failure by disturbing decidualization of ESCs via the miR-3074-5p/CLN8 pathway, providing a potential diagnostic and therapeutic target for RM.

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Qian Zhang State Key Laboratory of Reproductive Biology, Department of Obstetrics, Key Laboratory of Longevity and Ageing-related Diseases, Laboratory Animal Center, School of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China
State Key Laboratory of Reproductive Biology, Department of Obstetrics, Key Laboratory of Longevity and Ageing-related Diseases, Laboratory Animal Center, School of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China

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Song Yu State Key Laboratory of Reproductive Biology, Department of Obstetrics, Key Laboratory of Longevity and Ageing-related Diseases, Laboratory Animal Center, School of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China

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Xing Huang State Key Laboratory of Reproductive Biology, Department of Obstetrics, Key Laboratory of Longevity and Ageing-related Diseases, Laboratory Animal Center, School of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China
State Key Laboratory of Reproductive Biology, Department of Obstetrics, Key Laboratory of Longevity and Ageing-related Diseases, Laboratory Animal Center, School of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China

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Yi Tan State Key Laboratory of Reproductive Biology, Department of Obstetrics, Key Laboratory of Longevity and Ageing-related Diseases, Laboratory Animal Center, School of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China

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Cheng Zhu State Key Laboratory of Reproductive Biology, Department of Obstetrics, Key Laboratory of Longevity and Ageing-related Diseases, Laboratory Animal Center, School of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China

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Yan-Ling Wang State Key Laboratory of Reproductive Biology, Department of Obstetrics, Key Laboratory of Longevity and Ageing-related Diseases, Laboratory Animal Center, School of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China

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Haibin Wang State Key Laboratory of Reproductive Biology, Department of Obstetrics, Key Laboratory of Longevity and Ageing-related Diseases, Laboratory Animal Center, School of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China

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Hai-Yan Lin State Key Laboratory of Reproductive Biology, Department of Obstetrics, Key Laboratory of Longevity and Ageing-related Diseases, Laboratory Animal Center, School of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China

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Jiejun Fu State Key Laboratory of Reproductive Biology, Department of Obstetrics, Key Laboratory of Longevity and Ageing-related Diseases, Laboratory Animal Center, School of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China

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Hongmei Wang State Key Laboratory of Reproductive Biology, Department of Obstetrics, Key Laboratory of Longevity and Ageing-related Diseases, Laboratory Animal Center, School of Life Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China

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Cullin 3 (CUL3), a scaffold protein, assembles a large number of ubiquitin ligase complexes, similar to Skp1-Cullin 1-F-box protein complex. Several genetic models have shown that CUL3 is crucial for early embryonic development. Nevertheless, the role of CUL3 in human trophoblast function remains unclear. In this study, immunostaining revealed that CUL3 was strongly expressed in the villous cytotrophoblasts, the trophoblast column, and the invasive extravillous trophoblasts. Silencing CUL3 significantly inhibited the outgrowth of villous explant ex vivo and decreased invasion and migration of trophoblast HTR8/SVneo cells. Furthermore, CUL3 siRNA decreased pro-MMP9 activity and increased the levels of TIMP1 and 2. We also found that the level of CUL3 in the placental villi from pre-eclamptic patients was significantly lower as compared to that from their gestational age-matched controls. Moreover, in the lentiviral-mediated placenta-specific CUL3 knockdown mice, lack of CUL3 resulted in less invasive trophoblast cells in the maternal decidua. Taken together, these results suggest an essential role for CUL3 in the invasion and migration of trophoblast cells, and dysregulation of its expression may be associated with the onset of pre-eclampsia.

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Xiaohui Cui Dairy Cattle Research Center, Shandong Academy of Agricultural Science, Jinan, People’s Republic of China
College of Life Science, Shandong Normal University, Jinan, People’s Republic of China

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Yan Sun Dairy Cattle Research Center, Shandong Academy of Agricultural Science, Jinan, People’s Republic of China

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Xiuge Wang Dairy Cattle Research Center, Shandong Academy of Agricultural Science, Jinan, People’s Republic of China

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Chunhong Yang Dairy Cattle Research Center, Shandong Academy of Agricultural Science, Jinan, People’s Republic of China

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Zhihua Ju Dairy Cattle Research Center, Shandong Academy of Agricultural Science, Jinan, People’s Republic of China

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Qiang Jiang Dairy Cattle Research Center, Shandong Academy of Agricultural Science, Jinan, People’s Republic of China

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Yan Zhang Dairy Cattle Research Center, Shandong Academy of Agricultural Science, Jinan, People’s Republic of China

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Jinming Huang Dairy Cattle Research Center, Shandong Academy of Agricultural Science, Jinan, People’s Republic of China

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Jifeng Zhong Dairy Cattle Research Center, Shandong Academy of Agricultural Science, Jinan, People’s Republic of China

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Miao Yin College of Life Science, Shandong Normal University, Jinan, People’s Republic of China

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Changfa Wang Dairy Cattle Research Center, Shandong Academy of Agricultural Science, Jinan, People’s Republic of China

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The micromolar calcium-activated neutral protease gene (CAPN1) is a physiological candidate gene for sperm motility. However, the molecular mechanisms involved in regulating the expression of the CAPN1 gene in bulls remain unknown. In this study, we investigated the expression pattern of CAPN1 in testis, epididymis, and sperm at the RNA and protein levels by qRT-PCR, western blot, immunohistochemistry, and immunofluorescence assay. Results revealed that the expression of CAPN1 levels was higher in the sperm head compared with that in other tissues. Moreover, we identified a novel single-nucleotide polymorphism (g.-1256 A>C, ss 1917715340) in the noncanonical core promoter of the CAPN1 gene between base g.-1306 and g.-1012. Additionally, we observed greater sperm motility in bulls with the genotype CC than in those with the genotype AA (P<0.01), indicating that different genotypes were associated with the bovine semen trait. Furthermore, a higher fluorescence intensity of the C allele than that of the A allele at g. -1256 A>C was revealed by transient transfection in MLTC-1 cells and luciferase report assay. Finally, CAPN1 was highly expressed in the spermatozoa with the CC genotype compared with that with the AA genotype by qRT-PCR. This study is the first report on genetic variant g.-1256 A>C in the promoter region of CAPN1 gene association with the semen quality of Chinese Holstein bulls by influencing its expression. g.-1256 A>C can be a functional molecular marker in cattle breeding.

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Hang Qi Department of Obstetrics and Gynecology, International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China
Institute of Embryo-Fetal Original Adult Disease, Affiliated to School of Medicine, Shanghai Jiaotong University, Shanghai, China

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Guiling Liang Department of Obstetrics and Gynecology, International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China
Institute of Embryo-Fetal Original Adult Disease, Affiliated to School of Medicine, Shanghai Jiaotong University, Shanghai, China

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Jin Yu Department of Obstetrics and Gynecology, International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China
Institute of Embryo-Fetal Original Adult Disease, Affiliated to School of Medicine, Shanghai Jiaotong University, Shanghai, China

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Xiaofeng Wang Department of Obstetrics and Gynecology, International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China
Institute of Embryo-Fetal Original Adult Disease, Affiliated to School of Medicine, Shanghai Jiaotong University, Shanghai, China

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Yan Liang Department of Obstetrics and Gynecology, International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China
Institute of Embryo-Fetal Original Adult Disease, Affiliated to School of Medicine, Shanghai Jiaotong University, Shanghai, China

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Xiaoqing He Department of Obstetrics and Gynecology, International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China
Institute of Embryo-Fetal Original Adult Disease, Affiliated to School of Medicine, Shanghai Jiaotong University, Shanghai, China

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Tienan Feng Clinical Research Center, Shanghai Jiao Tong University School of Medicine, Shanghai, China

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Jian Zhang Department of Obstetrics and Gynecology, International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China
Institute of Embryo-Fetal Original Adult Disease, Affiliated to School of Medicine, Shanghai Jiaotong University, Shanghai, China

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MicroRNA (miRNA) expression profiles in tubal endometriosis (EM) are still poorly understood. In this study, we analyzed the differential expression of miRNAs and the related gene networks and signaling pathways in tubal EM. Four tubal epithelium samples from tubal EM patients and five normal tubal epithelium samples from uterine leiomyoma patients were collected for miRNA microarray. Bioinformatics analyses, including Ingenuity Pathway Analysis (IPA), Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, were performed. Quantitative real-time polymerase chain reaction (qRT-PCR) validation of five miRNAs was performed in six tubal epithelium samples from tubal EM and six from control. A total of 17 significantly differentially expressed miRNAs and 4343 potential miRNA-target genes involved in tubal EM were identified (fold change >1.5 and FDR-adjusted P value <0.05). IPA indicated connections between miRNAs, target genes and other gynecological diseases like endometrial carcinoma. GO and KEGG analysis revealed that most of the identified genes were involved in the mTOR signaling pathway, SNARE interactions in vesicular transport and endocytosis. We constructed an miRNA-gene-disease network using target gene prediction. Functional analysis showed that the mTOR pathway was connected closely to tubal EM. Our results demonstrate for the first time the differentially expressed miRNAs and the related signal pathways involved in the pathogenesis of tubal EM which contribute to elucidating the pathogenic mechanism of tubal EM-related infertility.

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Kang Shan Department of Obstetrics and Gynecology, Fourth Hospital, Hebei Medical University, Shijiazhuang 050011, China, Department of Molecular Biology, Hebei Cancer Institute, Hebei Medical University, Jiankanglu 12, Shijiazhuang 050011, China and Department of Epidemiology, Hebei Cancer Institute, Shijiazhuang 050011, China

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Ma Xiao-Wei Department of Obstetrics and Gynecology, Fourth Hospital, Hebei Medical University, Shijiazhuang 050011, China, Department of Molecular Biology, Hebei Cancer Institute, Hebei Medical University, Jiankanglu 12, Shijiazhuang 050011, China and Department of Epidemiology, Hebei Cancer Institute, Shijiazhuang 050011, China

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Wang Na Department of Obstetrics and Gynecology, Fourth Hospital, Hebei Medical University, Shijiazhuang 050011, China, Department of Molecular Biology, Hebei Cancer Institute, Hebei Medical University, Jiankanglu 12, Shijiazhuang 050011, China and Department of Epidemiology, Hebei Cancer Institute, Shijiazhuang 050011, China

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Zhang Xiu-Feng Department of Obstetrics and Gynecology, Fourth Hospital, Hebei Medical University, Shijiazhuang 050011, China, Department of Molecular Biology, Hebei Cancer Institute, Hebei Medical University, Jiankanglu 12, Shijiazhuang 050011, China and Department of Epidemiology, Hebei Cancer Institute, Shijiazhuang 050011, China

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Wen Deng-Gui Department of Obstetrics and Gynecology, Fourth Hospital, Hebei Medical University, Shijiazhuang 050011, China, Department of Molecular Biology, Hebei Cancer Institute, Hebei Medical University, Jiankanglu 12, Shijiazhuang 050011, China and Department of Epidemiology, Hebei Cancer Institute, Shijiazhuang 050011, China

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Guo Wei Department of Obstetrics and Gynecology, Fourth Hospital, Hebei Medical University, Shijiazhuang 050011, China, Department of Molecular Biology, Hebei Cancer Institute, Hebei Medical University, Jiankanglu 12, Shijiazhuang 050011, China and Department of Epidemiology, Hebei Cancer Institute, Shijiazhuang 050011, China

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Zhang Zheng-Mao Department of Obstetrics and Gynecology, Fourth Hospital, Hebei Medical University, Shijiazhuang 050011, China, Department of Molecular Biology, Hebei Cancer Institute, Hebei Medical University, Jiankanglu 12, Shijiazhuang 050011, China and Department of Epidemiology, Hebei Cancer Institute, Shijiazhuang 050011, China

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Li Yan Department of Obstetrics and Gynecology, Fourth Hospital, Hebei Medical University, Shijiazhuang 050011, China, Department of Molecular Biology, Hebei Cancer Institute, Hebei Medical University, Jiankanglu 12, Shijiazhuang 050011, China and Department of Epidemiology, Hebei Cancer Institute, Shijiazhuang 050011, China

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Endometriosis, one of the most frequent diseases in gynecology, is a benign but invasive and metastatic disease. The altered expression of E-cadherin may play an important role in developing endometriosis. In this paper, we discuss the association of three single nucleotide polymorphisms (SNPs) on the E-cadherin gene and risk of endometriosis. We examined the genotype frequency of three polymorphisms in 152 endometriosis patients and 189 control women. There was a significant difference in the frequency of the E-cadherin 3′-UTR C → T genotypes between endometriosis and controls (P = 0.01). The frequency of the C allele in patients (71.1%) was significantly higher than in the controls (63.8%; P = 0.04). When compared with the T/T + T/C genotypes, the C/C genotype had a significantly increased susceptibility to endometriosis, with an adjusted odds ratio of 1.79 (95% confidence interval = 1.17–2.76). No significant difference was found between endometriosis and control women on two polymorphisms (−160 C → A, −347 G → GA) at the gene promoter region of E-cadherin. The −160 C → A and −347 G → GA polymorphisms displayed linkage disequilibrium (D′ = 0.999). The −160 A/−347 GA haplotype was only detected in endometriosis patients (2%). These data show a relation between the E-cadherin 3′-UTR C → T polymorphism, the −160 A/−347 GA haplotype of two promoter polymorphisms and risk of endometriosis, suggesting a potential role in endometriosis development, at least in North Chinese women.

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Ma Tian-Zhong Reproductive Medicine Center, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong, China

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Chen Bi Reproductive Medicine Center, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong, China

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Zhang Ying Department of Gynecology and Obstetrics, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong, China

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Jing Xia Reproductive Medicine Center, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong, China

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Peng Cai-Ling Reproductive Medicine Center, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong, China

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Zhang Yun-Shan Reproductive Medicine Center, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong, China

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Huang Mei-Wen Reproductive Medicine Center, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong, China

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Niu Yan-Ru Laboratory of Minimally Invasive Orthopaedic, Guangdong Medical College, Zhanjiang, Guangdong, China

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Abstract

Emx2 deletion impairs the growth and maintenance of the genital ridge. However, its role in subsequent germ cell differentiation during embryonic stages is unknown. Using a tamoxifen-inducible Cre-loxP mouse model (Emx2 flox/flox, Cre-ER TM, hereafter called as Emx2 knockdown), we showed that germ cell differentiation was impaired in Emx2-knockdown testes. Representative characteristics of male germ cell differentiation, including a reduced ability to form embryonic germ (EG) cell colonies in vitro, down-regulation of pluripotency markers and G1/G0 arrest, did not occur in Emx2-knockdown testes. Furthermore, FGF9 and NODAL signalling occurred at abnormally high levels in Emx2-knockdown testes. Both blocking FGF9 signalling with SU5402 and inhibiting NODAL signalling with SB431542 allowed germ cells from Emx2-knockdown testes to differentiate in vitro. Therefore, EMX2 in somatic cells is required to trigger germ cell differentiation in XY foetuses, posterior to its previously reported role in the growth and maintenance of the genital ridge.

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Miaomiao Jin College of Veterinary Medicine, Northwest A&F University, Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Yangling, Shaanxi, China

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Lu Zhao College of Veterinary Medicine, Northwest A&F University, Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Yangling, Shaanxi, China

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Hanwen Yang College of Veterinary Medicine, Northwest A&F University, Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Yangling, Shaanxi, China

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Jianglin Zhao College of Veterinary Medicine, Northwest A&F University, Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Yangling, Shaanxi, China

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Hongwei Ma College of Veterinary Medicine, Northwest A&F University, Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Yangling, Shaanxi, China

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Yanzhi Chen College of Veterinary Medicine, Northwest A&F University, Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Yangling, Shaanxi, China

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Jingcheng Zhang College of Veterinary Medicine, Northwest A&F University, Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Yangling, Shaanxi, China

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Yan Luo College of Animal Engineering, Yangling Vocational and Technical College, Yangling, Shaanxi, China

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Yong Zhang College of Veterinary Medicine, Northwest A&F University, Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Yangling, Shaanxi, China

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Jun Liu College of Veterinary Medicine, Northwest A&F University, Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Yangling, Shaanxi, China

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In brief

Early embryonic development in goats is a complex and an important process. This study identified a novel long non-coding RNA (lncRNA), lncRNA3720, that appears to affect early embryonic development in goats through histone variants.

Abstract

Although abundant lncRNAs have been found to be highly expressed in early embryos, the functions and mechanisms of most lncRNAs in regulating embryonic development remain unclear. This study was conducted to identify the key lncRNAs during embryonic genome activation (EGA) for promoting embryonic development after somatic cell nuclear transfer (SCNT) in goats. We screened and characterized lncRNAs from transcriptome data of in vitro-fertilized, two-cell (IVF-2c) and eight-cell embryos (IVF-8c) and eight-cell SCNT embryos (SCNT-8c). We obtained 12 differentially expressed lncRNAs that were highly expressed in IVF-8c embryos compared to IVF-2c and less expressed in SCNT-8c embryos. After target gene prediction, expression verification, and functional deletion experiments, we found that the expression level of lncRNA3720 affected the early embryonic development in goats. We cloned full-length lncRNA3720 and over-expressed it in goat fetal fibroblasts (GFFs). We identified histone variants by analyzing the transcriptome data from both GFFs and embryos. Gene annotation of the gene library and the literature search revealed that histone variants may have important roles in early embryo development, so we selected them as the potential target genes for lncRNA3720. Lastly, we compensated for the low expression of lncRNA3720 in SCNT embryos by microinjection and showed that the development rate and quality of SCNT embryos were significantly improved. We speculate that lncRNA3720 is a key promoter of embryonic development in goats by interacting with histone variants.

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Mingxi Yu Laboratory of Animal Cell and Molecular Biology, College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang, Liaoning, China

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Xinlong Jiang Laboratory of Animal Cell and Molecular Biology, College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang, Liaoning, China

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Wenyang Cai Laboratory of Animal Cell and Molecular Biology, College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang, Liaoning, China

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Xiao Yang Laboratory of Animal Cell and Molecular Biology, College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang, Liaoning, China

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Wenzhong An Laboratory of Animal Cell and Molecular Biology, College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang, Liaoning, China

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Mei Zhang Laboratory of Animal Cell and Molecular Biology, College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang, Liaoning, China

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Yan Xu Laboratory of Animal Cell and Molecular Biology, College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang, Liaoning, China

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Bochuan Zhang Laboratory of Animal Cell and Molecular Biology, College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang, Liaoning, China

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Shuang Tang Laboratory of Animal Cell and Molecular Biology, College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang, Liaoning, China

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In brief

During the morula to blastocyst transformation, polarity establishment in outer cells is a prerequisite for trophectoderm lineage specification. This study reveals the roles of polarity proteins PATJ and MPDZ in trophectoderm lineage fate decision.

Abstract

In mouse preimplantation embryos, cell polarity plays a crucial role in the first lineage specification. PATJ and its homolog MPDZ are the main members of CRB–PALS1–PATJ (CRUMBS–Protein associated with Lin7 1–Pals-associated tight junction protein) apical polarity complex. They act as adaptor proteins connecting CRB–PALS1 and tight junction proteins, making them essential for cell polarization and stabilization of apical junctions. However, their roles in regulating trophectoderm differentiation and blastocyst development remain unclear. In this study, PATJ and/or MPDZ were downregulated by the microinjection of specific RNA interference constructs into zygotes. Downregulation of PATJ alone did not severely affect early embryonic development and trophectoderm lineage differentiation although it slowed down the blastocyst formation. Depletion of PATJ and MPDZ did not affect compaction and morula development but impaired blastocyst formation. Furthermore, the expression of trophectoderm-specific transcription factors and trophoblast differentiation was compromised in the absence of PATJ/MPDZ. These abnormalities might result from the breakdown of apical domain in the outer cells of the embryo. The loss of PATJ/MPDZ caused the breakdown of CRB and PAR polarity complexes as well as deficiencies in tight junctions and actin filaments. These defects led to ectopic activation of Hippo signaling in the outer cells of developing embryos, ultimately suppressing Cdx2 expression and trophectoderm differentiation. Altogether, PATJ and MPDZ are essential for trophectoderm lineage differentiation and normal blastocyst morphogenesis via the regulation of the establishment of apical domain, formation of tight junctions, phosphorylation and localization of YAP, and expression of trophectoderm-specific transcription factors.

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