INCREASED TESTICULAR CD2+ & INFERTILITY WITH VARICOCELE

睾丸 CD2 增加

• 批准号: 6382367
• 负责人: SUSAN H BENOFF
• 金额: $ 30.88万
• 依托单位: NORTH SHORE UNIVERSITY HOSPITAL
• 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/6382367
• 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 美国 20% 以上的男性不育症患者。手术修复 精索静脉曲张恢复生育能力的时间只有 1/3。 VAI 的识别可能 手术矫正的好处可以节省健康成本。在初步 研究表明，VAI 患者的精浆和睾丸活检中含有显着 镉 (Cd2+) 含量高于其他不育男性，尽管血液中的镉含量 无法区分。 VAI 精子失去正常的精子表面蛋白结合肌动蛋白， 包括甘露糖和黄体酮受体以及正常精子头 细胞骨架结构。 Cd2+ 中人类可育精子的体外培养 含有复制 VAI 细胞骨架缺陷的培养基。自动金相成像 的活检证实 VAI 睾丸中存在高 Cd2+。探测连续睾丸部分 研究表明肌动蛋白表达随分化阶段减少，并增加 VAI 小管中凋亡细胞核的频率。精子电压依赖性钙 通道 (VDCC) 是 Cd2+ VDCC 同种型表达的明显入口点 变异性与甘露糖受体功能相关。 VAI 患者合作伙伴 只有手术矫正恢复正常后三年内才能怀孕 甘露糖受体功能。因此Cd2+可能是热量的辅助因子 生产奥钢联。在一项扩大的前瞻性研究中，我们将确定是否具有开创性 Cd2+ 与睾丸 Cd2+ 相关，如果精液 Cd2+，则睾丸 Cd2+ 和 阴囊温度是通过怀孕和评估来评估 VAI 逆转的生物标志物 出生前，进行一次睾丸活检并复制精液样本 3 修正后几个月。用于表征 VAI 的中间端点将 包括：精浆和睾丸Zn2+、睾丸Cd2+和Zn2+ 自动金相学分布、精子对赋能孵化的反应 （甘露糖受体测定）和多价顶体反应诱导 甘露糖，评估肌动蛋白分布、细胞凋亡水平和 VDCC 表达。对照组将包括接受 IVF/ICSI 治疗的夫妇中的男性 非梗阻性无精子症和患有原发性 VAI 且拒绝矫正的男性。 为了进一步表征 VAI，中间终点的异常 个体（例如细胞凋亡和肌动蛋白丢失）将与 影响金属离子敏感性、通道区域的 VDCC 一级结构 根据 VDCC mRNA 评估门控电位和 Ca2+ 通道阻断剂结合 存在于成熟精子中，并且 VDCC 通道前体区域发生变化 通过精子蛋白质印迹评估对甘露糖诱导的蛋白水解敏感 膜蛋白。对照将包括具有生育能力的捐赠者和具有生育能力的男性 精索静脉曲张。