Stattic V, a STAT3 inhibitor, affects human spermatozoa through regulation of mitochondrial activity
Stattic V, a STAT3 inhibitor, affects human spermatozoa through regulation of mitochondrial activity
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DOI：
10.1002/jcp.24215
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发表时间：
2013-04-01
影响因子：
5.6
通讯作者：
Leclerc, Pierre
中科院分区：
文献类型：
--
作者：
Lachance, Catherine;Goupil, Serge;Leclerc, Pierre
我们最近发现许多JAK/STAT信号通路的介质存在于人类射精精子中。其中，STAT3主要在细胞膜和鞭毛细胞骨架部分检测到。为了确定stat3介导的信号的重要性，精子与Stattic V（一种特异性抑制剂）一起孵育。采用CASA法检测精子活力，采用FITC偶联凝集素（PSA或PNA）染色法检测精子顶体完整性，采用单克隆抗磷酸酪氨酸抗体Western blot法检测蛋白磷酸酪氨酸含量。用INDO1-AM和JC-1分别流式细胞术检测精子细胞内钙和线粒体膜电位，用发光胺法检测活性氧（ROS）的产生。静态v显著影响运动百分比和运动参数，随后也显著增加细胞内Ca2+水平，黄体酮和钙离子载体（A23187）诱导顶体反应。另一方面，用statstatic V处理精子时，检测到ATP含量显著降低，这与线粒体膜去极化和ROS生成升高有关。这些结果表明，STAT3至少通过调节线粒体活性参与精子功能。这进一步强调了STAT3介导细胞活动的方式不同于严格意义上的基因转录激活。j .细胞。中国生物医学工程学报，2016,33(2):444 - 444。(c) 2012 Wiley期刊有限公司
We have recently shown that many mediators of the JAK/STAT signaling pathway are present in ejaculated human spermatozoa. Among them, STAT3 is detected mainly in membranes and flagellar cytoskeletal fractions. In order to determine the importance of STAT3-mediated signaling, sperm were incubated with Stattic V, a specific inhibitor. Effects on motility were evaluated by CASA, sperm acrosomal integrity was evaluated by FITC conjugated lectin (PSA or PNA) staining, and protein phosphotyrosine content was assessed by Western blot using a monoclonal anti-phosphotyrosine antibody. INDO1-AM and JC-1 were used to measure sperm intracellular calcium and mitochondrial membrane potential, respectively, by flow cytometry, and reactive oxygen species (ROS) production was investigated by luminol-based assay. Percentages of motility and motility parameters were significantly affected by Stattic V. This later also significantly increased intracellular Ca2+ levels, progesterone- and calcium ionophore (A23187)-induced acrosome reaction. On the other hand, a significant decrease in ATP content was measured when sperm were treated with Stattic V, associated with depolarization of mitochondrial membrane and elevated ROS production. These results suggest that STAT3 is involved in sperm functions, at least through regulation of mitochondrial activity. This further emphasizes that STAT3 mediates cellular activities in a manner different than strictly the activation of gene transcription. J. Cell. Physiol. 228: 704713, 2013. (c) 2012 Wiley Periodicals, Inc.