Heat shock protein 90 has roles in intracellular calcium homeostasis, protein tyrosine phosphorylation regulation, and progesterone-responsive sperm function in human sperm.
Heat shock protein 90 has roles in intracellular calcium homeostasis, protein tyrosine phosphorylation regulation, and progesterone-responsive sperm function in human sperm.
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热休克蛋白 90 在人类精子的细胞内钙稳态、蛋白酪氨酸磷酸化调节和孕酮反应性精子功能中发挥作用
DOI：
10.1371/journal.pone.0115841
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Ni Y
中科院分区：
文献类型：
--
作者：
Li K;Xue Y;Chen A;Jiang Y;Xie H;Shi Q;Zhang S;Ni Y
热休克蛋白90在客户蛋白的成熟、信号转导、蛋白质折叠和降解以及形态进化中起着关键作用，但其在人类精子中的功能尚不完全清楚。因此，本研究的目的是阐明热休克蛋白90对人类精子功能的影响机制。通过间接免疫荧光染色，我们发现热休克蛋白90主要定位于人精子的颈部、中段和尾部，并且在获能条件下，其表达随着孵育时间的延长而增加。Western blotting分析表明，热休克蛋白90的特异性抑制剂格尔达那霉素可以抑制热休克蛋白90表达的增加。使用多功能微孔板阅读器检测Fluo-3AM负载的精子，我们首次观察到在获能过程中使用格尔达霉素抑制热休克蛋白90显著降低细胞内钙离子浓度。此外，Western印迹分析表明，格尔达霉素以剂量依赖的方式促进了包括热休克蛋白90在内的几种蛋白质的酪氨酸磷酸化。通过金霉素染色和计算机辅助精子分析仪，评估了在没有或存在黄体酮的情况下，格尔达霉素对人类精子功能的影响。我们发现，格尔达那霉素本身并不影响精子的获能、超激活和活力，但在黄体酮存在的情况下会影响精子的获能、超激活和运动能力。综上所述，这些数据表明，热休克蛋白90在获能过程中在人类精子中的表达增加，在细胞内钙稳态、蛋白酪氨酸磷酸化调节和黄体酮刺激的精子功能中发挥作用。在这项研究中，我们对热休克蛋白90在精子功能中的作用提供了新的见解。
Heat shock protein 90 plays critical roles in client protein maturation, signal transduction, protein folding and degradation, and morphological evolution; however, its function in human sperm is not fully understood. Therefore, our objective in this study was to elucidate the mechanism by which heat shock protein 90 exerts its effects on human sperm function. By performing indirect immunofluorescence staining, we found that heat shock protein 90 was localized primarily in the neck, midpiece, and tail regions of human sperm, and that its expression increased with increasing incubation time under capacitation conditions. Geldanamycin, a specific inhibitor of heat shock protein 90, was shown to inhibit this increase in heat shock protein 90 expression in western blotting analyses. Using a multifunctional microplate reader to examine Fluo-3 AM-loaded sperm, we observed for the first time that inhibition of heat shock protein 90 by using geldanamycin significantly decreased intracellular calcium concentrations during capacitation. Moreover, western blot analysis showed that geldanamycin enhanced tyrosine phosphorylation of several proteins, including heat shock protein 90, in a dose-dependent manner. The effects of geldanamycin on human sperm function in the absence or presence of progesterone was evaluated by performing chlortetracycline staining and by using a computer-assisted sperm analyzer. We found that geldanamycin alone did not affect sperm capacitation, hyperactivation, and motility, but did so in the presence of progesterone. Taken together, these data suggest that heat shock protein 90, which increases in expression in human sperm during capacitation, has roles in intracellular calcium homeostasis, protein tyrosine phosphorylation regulation, and progesterone-stimulated sperm function. In this study, we provide new insights into the roles of heat shock protein 90 in sperm function.
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DOI：
10.1073/pnas.0605795103
发表时间：
2006-10-10
影响因子：
11.1
作者：
Gomendio, Montserrat;Martin-Coello, Juan;Roldan, Eduardo R. S.
通讯作者：
Roldan, Eduardo R. S.
影响因子：
4.8
作者：
Alekseev, OM;Widgren, EE;O'Rand, MG
通讯作者：
O'Rand, MG
影响因子：
3.5
作者：
ITOH, H;TASHIMA, Y
通讯作者：
TASHIMA, Y
影响因子：
3.7
作者：
Dai B;Wang Y;Li D;Xu Y;Liang R;Zhao L;Cao Y;Jia J;Jiang Y
通讯作者：
Jiang Y
影响因子：
10.5
作者：
Gur, Y;Breitbart, H
通讯作者：
Breitbart, H