INCREASED TESTICULAR CD2+ & INFERTILITY WITH VARICOCELE

睾丸 CD2 增加

• 批准号: 6635513
• 负责人: SUSAN H BENOFF
• 金额: $ 30.34万
• 依托单位: FEINSTEIN INSTITUTE FOR MEDICAL RESEARCH
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-05-01 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6635513
• 关键词: actins; biopsy; body temperature; cadmium; clinical research; fertility; human subject; male; semen; sperm; testis disorder

DESCRIPTION: Varicocele-associated human male infertility (VAI) represents greater then 20 percent of male infertility in the US. Surgical repair of varicocele restores fertility only 1/3 of time. Identification of VAI likely to benefit from surgical correction offers health cost savings. In preliminary studies, VAI patients' seminal plasma and testes biopsies contain significantly higher cadmium (Cd2+) than other infertile males, though blood levels were indistinguishable. VAI sperm lose normal sperm surface proteins binding actin, including mannose and progesterone receptors, and normal sperm head cytoskeletal structures. In vitro incubation of fertile human sperm in Cd2+ containing media replicate VAI cytoskeletal defects. Autometallographic imaging of biopsies confirmed high Cd2+ in VAI testes. Probing serial testes sections revealed decreased actin expression with differentiation stage, and increased frequency of apoptotic nuclei in VAI tubules. Sperm voltage-dependent calcium channels (VDCC) are an apparent entry point for Cd2+ VDCC isoform expression variability correlates with mannose receptor function. VAI patient partners became pregnant within three years only if surgical correction restored normal mannose receptor function. Therefore Cd2+ may be a cofactor with heat in producing VAI. In an expanded prospective study we will determine if seminal Cd2+ correlates with testicular Cd2+, and if seminal Cd2+, testicular Cd2+ and scrotal temperature are biomarkers for VAI reversal assessed by pregnancy and birth, taking one testicular biopsy and duplicate semen samples before and 3 months after correction. Intermediate endpoints for characterizing VAI will include: seminal plasma and testicular Zn2+, testicular Cd2+ and Zn2+ distributions by autometallography, sperm response to capacitating incubation (mannose receptor assay) and to acrosome-reaction induction by polvalent mannose, assessment of actin distributions, levels of apoptosis and VDCC expression. Controls will include men from couples undergoing IVF/ICSI for non-obstructive azoospermia and males with primary VAI who refuse correction. To further characterize VAI, abnormalities in intermediate endpoints among individuals (e.g. apoptosis and actin loss) will be correlated with changes in VDCC primary structure in regions afecting metal ion sensitivity, channel gating potential and Ca2+ channel blocker binding as assesed from VDCC mRNA present in mature sperm, and with changes in a region of VDCC channel precursor sensitive to mannose-induced proteolysis, assessed by Western blot of sperm membrane proteins. Controls will include fertile donors and fertile men with varicocele.

描述：精索静脉曲张相关的人类男性不育症（VAI）代表 超过20%的美国男性不育。手术修复 精索静脉曲张恢复生育能力的时间只有1/3。确定可能 从手术矫正中获益可节省医疗费用。初步 研究表明，VAI患者的精浆和睾丸活检中含有显著的 高镉（Cd 2+）比其他不育男性，虽然血液水平 无法区分VAI精子失去了正常的精子表面蛋白结合肌动蛋白， 包括甘露糖和孕酮受体，以及正常的精子头 细胞骨架结构人生育精子在Cd ~（2+）中的体外孵育 含有培养基复制VAI细胞骨架缺陷。自显影成像 的活组织检查证实VAI睾丸中存在高Cd 2+。探测连续睾丸切片 肌动蛋白表达随着分化阶段的增加而降低， VAI小管中凋亡核的频率。精子电压依赖性钙 通道（VDCC）是Cd 2 + VDCC亚型表达的明显切入点 变异性与甘露糖受体功能相关。奥钢联患者合作伙伴 只有手术矫正恢复正常才能在三年内怀孕 甘露糖受体功能因此，Cd 2+可能是一个辅因子与热在 生产VAI。在一项扩大的前瞻性研究中，我们将确定 Cd ~（2+）与睾丸Cd ~（2+）相关，如果精液Cd ~（2+）、睾丸Cd ~（2+）和 阴囊温度是通过妊娠评估VAI逆转的生物标志物， 分娩前和分娩后3个月， 纠正后的几个月。表征VAI的中间终点将 包括精浆和睾丸Zn ~（2+），睾丸Cd ~（2+）和Zn ~（2 + 精子分布（自动记录法），精子对获能孵育的反应 （甘露糖受体测定）和顶体反应诱导的多价 甘露糖，肌动蛋白分布评估，凋亡水平和VDCC 表情对照将包括来自接受IVF/ICSI的夫妇的男性， 非梗阻性无精子症和拒绝矫正的原发性VAI男性。 为了进一步表征VAI， 个体（例如细胞凋亡和肌动蛋白损失）将与 影响金属离子灵敏度的区域中的VDCC一级结构，通道 根据VDCC mRNA评估的门控电位和Ca 2+通道阻滞剂结合 存在于成熟精子中，并且VDCC通道前体区域发生变化 对甘露糖诱导的蛋白水解敏感，通过精子的Western印迹评估 膜蛋白对照将包括有生育能力的供体和有生育能力的男性， 精索静脉曲张