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Hua Mei Reproductive Physiology Group, Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge CB2 3EG, UK

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Cara Walters Reproductive Physiology Group, Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge CB2 3EG, UK

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Richard Carter Reproductive Physiology Group, Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge CB2 3EG, UK

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William H Colledge Reproductive Physiology Group, Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge CB2 3EG, UK

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Mice with mutations in the kisspeptin signaling pathway (Kiss1 −/− or Gpr54 −/− ) have low gonadotrophic hormone levels, small testes, and impaired spermatogenesis. Between 2 and 7 months of age, however, the testes of the mutant mice increase in weight and in Gpr54 −/− mice, the number of seminiferous tubules containing spermatids/spermatozoa increases from 17 to 78%. In contrast, the Kiss1 −/− mice have a less severe defect in spermatogenesis and larger testes than Gpr54 −/− mice at both 2 and 7 months of age. The reason for the improved spermatogenesis was investigated. Plasma testosterone and FSH levels did not increase with age in the mutant mice and remained much lower than in wild-type (WT) mice. In contrast, intratesticular testosterone levels were similar between mutant and WT mice. These data indicate that age-related spermatogenesis can be completed under conditions of low plasma testosterone and FSH and that intratesticular testosterone may contribute to this process. In addition, however, when the Gpr54 −/− mice were fed a phytoestrogen-free diet, they showed no age-related increase in testes weight or improved spermatogenesis. Thus, both genetic and environmental factors are involved in the improved spermatogenesis in the mutant mice as they age although the mice still remain infertile. These data show that the possible impact of dietary phytoestrogens should be taken into account when studying the phenotype of mutant mice with defects in the reproductive axis.

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Yan Xu Medical School of Fudan University, Center for Biomedical Research, The Rockefeller University, National Population and Family Planning Key Laboratory of Contraceptive Drugs and Devices, Department of Anatomy, Shanghai, China

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Miao Liu Medical School of Fudan University, Center for Biomedical Research, The Rockefeller University, National Population and Family Planning Key Laboratory of Contraceptive Drugs and Devices, Department of Anatomy, Shanghai, China

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Yi-hua Gu Medical School of Fudan University, Center for Biomedical Research, The Rockefeller University, National Population and Family Planning Key Laboratory of Contraceptive Drugs and Devices, Department of Anatomy, Shanghai, China

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Xiao-feng Jia Medical School of Fudan University, Center for Biomedical Research, The Rockefeller University, National Population and Family Planning Key Laboratory of Contraceptive Drugs and Devices, Department of Anatomy, Shanghai, China

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Yong-Mei Chen Medical School of Fudan University, Center for Biomedical Research, The Rockefeller University, National Population and Family Planning Key Laboratory of Contraceptive Drugs and Devices, Department of Anatomy, Shanghai, China

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Michelle Santos Medical School of Fudan University, Center for Biomedical Research, The Rockefeller University, National Population and Family Planning Key Laboratory of Contraceptive Drugs and Devices, Department of Anatomy, Shanghai, China

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Ai-Zhen Wu Medical School of Fudan University, Center for Biomedical Research, The Rockefeller University, National Population and Family Planning Key Laboratory of Contraceptive Drugs and Devices, Department of Anatomy, Shanghai, China

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Xiao-dong Zhang Medical School of Fudan University, Center for Biomedical Research, The Rockefeller University, National Population and Family Planning Key Laboratory of Contraceptive Drugs and Devices, Department of Anatomy, Shanghai, China

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Hui-Juan Shi Medical School of Fudan University, Center for Biomedical Research, The Rockefeller University, National Population and Family Planning Key Laboratory of Contraceptive Drugs and Devices, Department of Anatomy, Shanghai, China
Medical School of Fudan University, Center for Biomedical Research, The Rockefeller University, National Population and Family Planning Key Laboratory of Contraceptive Drugs and Devices, Department of Anatomy, Shanghai, China

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Ching-Ling C Chen Medical School of Fudan University, Center for Biomedical Research, The Rockefeller University, National Population and Family Planning Key Laboratory of Contraceptive Drugs and Devices, Department of Anatomy, Shanghai, China
Medical School of Fudan University, Center for Biomedical Research, The Rockefeller University, National Population and Family Planning Key Laboratory of Contraceptive Drugs and Devices, Department of Anatomy, Shanghai, China

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With tetraspanning topology, members of the membrane-spanning four-domain subfamily A (MS4A) may facilitate signaling or ion channel functions in many tissues. In this study, we report the cloning of a full-length cDNA from rat testis, designated Ms4a14 (Sp3111), which encodes the MS4A protein with 1139 amino acid residues. In situ hybridization and immunohistochemical analyses indicate that Ms4a14 is predominantly expressed from round spermatids to spermatozoa at specific stages in the rat testis at both the mRNA and protein level. Immunofluorescence analysis revealed that MS4A14 (SP3111) is located in the acrosome and the midpiece of the flagellum in mature sperm. Previously, we explored and reported the involvement of MS4A14 in reproductive functions, using antibody blockage during IVF and a transgenic RNA interference method in a mouse model. Our results suggested that MS4A14 is involved in fertilization and zygote division. As MS4A14 protein exists in mammals, such as humans, cows, dogs, and rodents, MS4A14 may play a ubiquitous role in mammalian reproduction.

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Xue-Ying Zhang The Key Laboratory of Reproductive Genetics (Zhejiang University), Ministry of Education, Hangzhou, Zhejiang, China

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Yi-Meng Xiong The Key Laboratory of Reproductive Genetics (Zhejiang University), Ministry of Education, Hangzhou, Zhejiang, China

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Ya-Jing Tan The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

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Li Wang The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

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Rong Li The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

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Yong Zhang The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

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Xin-Mei Liu The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

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Xian-Hua Lin The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

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Li Jin The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

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Yu-Ting Hu The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

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Zhen-Hua Tang The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

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Zheng-Mu Wu The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

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Feng-Hua Yin The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

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Zheng-Quan Wang The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

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Ye Xiao The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

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Jian-Zhong Sheng The Key Laboratory of Reproductive Genetics (Zhejiang University), Ministry of Education, Hangzhou, Zhejiang, China
Department of Pathology and Pathophysiology, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China

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He-Feng Huang The Key Laboratory of Reproductive Genetics (Zhejiang University), Ministry of Education, Hangzhou, Zhejiang, China
The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China
Shanghai Key Laboratory of Embryo Original Diseases, Shanghai, China

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Fertilization failure often occurs during in vitro fertilization (IVF) cycles despite apparently normal sperm and oocytes. Accumulating evidence suggests that mitochondria play crucial roles in the regulation of sperm function and male fertility. 3-Nitrophthalic acid (3-NPA) can induce oxidative stress in mitochondria, and melatonin, as an antioxidant, can improve mitochondrial function by reducing mitochondrial oxidative stress. The role of sperm mitochondrial dysfunction in fertilization failure during IVF is unclear. The present study revealed that spermatozoa with low, or poor, fertilization rates had swollen mitochondria, increased mitochondria-derived ROS, and attenuated mitochondrial respiratory capacity. 3-NPA treatment enhanced mitochondrial dysfunction in sperm. Spermatozoa with poor fertilization rates, and spermatozoa treated with 3-NPA, had reduced penetration ability. The concentration of melatonin was decreased in semen samples with low and poor fertilization rates. Melatonin, not only decreased excessive mitochondria-derived ROS, but also ‘rescued’ the reduced penetration capacity of spermatozoa treated with 3-NPA. Taken together, the study suggested that mitochondria-derived ROS and mitochondrial respiratory capacity are independent bio-markers for sperm dysfunction, and melatonin may be useful in improving sperm quality and overall male fertility.

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Wang Han-zheng
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Lu Shu-hua
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Shen Wei-xiong
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Sun Zhi-da
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Zhou Wei
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Wu Yu-fen
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Zhou Mei-rong
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Summary. Cell suspensions were prepared from human corpora lutea obtained during the mid-luteal phase. Progesterone production was assessed after short-term incubation of luteal cell suspensions. Luteal cells were very sensitive to hCG, the concentration required for 50% maximum response being 0·01 i.u./ml, and the response was 5 times higher than the basal production.

Oestradiol (1–100 μm) induced a significant dose-related decrease in both basal and hCG-stimulated progesterone production. The A-nor steroidal compounds anordrin and AF-45 reduced hCG-stimulated progesterone production only at the high concentration of 100 μm. The ED50 values were approximately 3 μm, 75 μm and 100 μm for oestradiol, AF-45 and anordrin respectively. Anordrin showed no significant effects on basal progesterone production. In addition, oestradiol markedly inhibited the activity of 3β-hydroxysteroid dehydrogenase in luteal cells, expressed by the conversion of pregnenolone to progesterone, but the inhibitory effects of anordrin and AF-45 were negligible or relatively low.

The effects of anordrin and AF-45 were different from those of oestradiol on progesterone production by human luteal cells in vitro, indicating that neither substance is likely to be a useful luteolytic agent in women.

Keywords: A-nor steroid; oestradiol; luteal cells; progesterone; 3β-hydroxysteroid dehydrogenase; man

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