Gonadotrophin-induced paracrine regulation of human oocyte maturation by BDNF and GDNF secreted by granulosa cells.
Gonadotrophin-induced paracrine regulation of human oocyte maturation by BDNF and GDNF secreted by granulosa cells.
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DOI：
10.1093/humrep/deq390
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发表时间：
2011-03
影响因子：
6.1
通讯作者：
Ping Zhao;J. Qiao;Shuo Huang;Ying Zhang;Shuang Liu;Liying Yan;A. Hsueh;E. Duan
中科院分区：
文献类型：
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作者：
Ping Zhao;J. Qiao;Shuo Huang;Ying Zhang;Shuang Liu;Liying Yan;A. Hsueh;E. Duan
在哺乳动物卵巢中，已经确定了多种旁分泌因子介导或调节lh诱导的排卵变化。由于体外受精过程中难以获得未受刺激的颗粒细胞，因此对人卵巢中lh诱导的旁分泌因子知之甚少。基于早期使用小鼠卵巢细胞的研究显示脑源性神经营养因子（BDNF）和胶质细胞系源性神经营养因子（GDNF）在促进卵母细胞成熟中的旁分泌作用，我们研究了这些配体在人颗粒细胞中的表达及其对人卵母细胞发育的调节。方法非刺激体外成熟患者取卵后获得非刺激颗粒细胞。接受非刺激IVM治疗的女性平均年龄为30.8±1.3岁（n = 10）。从接受促性腺激素刺激和ICSI的IVF患者收集未成熟卵母细胞和颗粒细胞。通过免疫细胞化学分析颗粒细胞BDNF和GDNF的表达谱，并结合实时RT-PCR分析促性腺激素对BDNF和GDNF转录物水平的调节。此外，培养未成熟卵母细胞，分析BDNF和GDNF对卵母细胞成熟的调节作用。结果BDNF和GDNF均在未受刺激的颗粒细胞中表达。促性腺激素（FSH和/或hCG）治疗后，BDNF和GDNF转录本水平显著升高（P < 0.05）。在培养的未成熟卵母细胞中，BDNF或GDNF处理可促进中期卵母细胞的总产量。结论：FSH和hCG可增强颗粒细胞BDNF和GDNF的表达，这些颗粒细胞来源的因子是能够促进卵母细胞成熟的候选旁分泌因子。
BACKGROUND In mammalian ovaries, diverse paracrine factors have been identified to mediate or modulate LH-induced changes during ovulation. Due to the difficulty in obtaining non-stimulated granulosa cells during IVF, little is known about the LH-induced paracrine factors in the human ovary. Based on earlier studies using murine ovarian cells showing the paracrine roles of brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) in promoting oocyte maturation, we investigated the expression of these ligands in human granulosa cells and their regulation of human oocyte development. METHODS Non-stimulated granulosa cells were obtained from non-stimulated IVM (in vitro maturation) patients after oocyte retrieval. Women undergoing non-stimulated IVM treatment at a mean age of 30.8 ± 1.3 (n = 10) were recruited for this study. Immature oocytes and granulosa cells were collected from IVF patients undergoing gonadotrophin stimulation and ICSI. Immunocytochemical analyses of granulosa cells were carried out to investigate expression profiles of BDNF and GDNF, together with real-time RT-PCR to analyze the gonadotrophin regulation of BDNF and GDNF transcript levels. In addition, immature oocytes were cultured to analyze the regulation of oocyte maturation by BDNF and GDNF. RESULTS BDNF and GDNF were found to be expressed in non-stimulated granulosa cells. After gonadotrophin (FSH and/or hCG) treatment, transcripts levels for BDNF and GDNF were significantly increased (P < 0.05). In cultured immature oocytes, treatment with BDNF or GDNF promoted total yields of metaphase II oocytes. CONCLUSIONS These findings demonstrate that FSH and hCG treatments augment the expression of BDNF and GDNF by granulosa cells and that these granulosa-cell-derived factors are candidate paracrine factors capable of promoting oocyte maturation.