Critical roles of Astrin in the mitosis of immature rat Sertoli cells.
Critical roles of Astrin in the mitosis of immature rat Sertoli cells.
复制标题
Astrin 在未成熟大鼠支持细胞有丝分裂中的关键作用。
DOI：
10.1016/j.bbrc.2017.03.137
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发表时间：
2017
影响因子：
3.1
通讯作者：
Hiroetsu Suzuki
中科院分区：
文献类型：
--
作者：
Yuki Tochigi;Yuka Iwasaki;Masanori Sano;Hidenori Yasuda;Kentaro Katayama;Hiroetsu Suzuki
雄性性腺功能减退（hgn/hgn）大鼠表现出睾丸发育不全并伴有精小管发育不良，这是由于编码Astrin的基因突变丧失功能导致的，Astrin是HeLa细胞分裂周期中有丝分裂进程所必需的。在本研究中，我们研究了导致hgnn / hgnes中支持细胞减少的细胞学基础。在出生后第3天，gata4阳性的Sertoli细胞中抗磷酸组蛋白H3 (Ser10) （pH3）阳性的有丝分裂期和tunel阳性的凋亡增加。从hgntestn中分离的未成熟Sertoli细胞显示ph3评估的有丝分裂指数升高，5-溴-2'-脱氧尿苷掺入减少，tunel阳性凋亡增加，提示有丝分裂延迟和细胞死亡。在nocodazole （NOC）介导的细胞周期阻滞和随后释放的有丝分裂过程可视化中，hgn/hgnrat衍生的Sertoli细胞未能从前期过渡到中期，微核细胞和tunel阳性细胞以时间依赖性的方式逐渐增加。Western blot分析在+/+和+/hgntestes提取物和培养的正常Sertoli细胞中检测到≈142 kDa蛋白，预计为Astrin，但在hgntestes提取物中未检测到。CLASP1在正常和hgnn / hgni睾丸提取物中均检测到，而在正常有丝分裂的支持细胞中定位于着丝点，而在hgnn /hgnSertoli细胞的细胞质中弥散。这些结果表明，Astrin是未成熟Sertoli细胞正常有丝分裂进程所必需的，而最严重的睾丸发育不良类型是由于缺乏Astrin导致未成熟Sertoli细胞有丝分裂细胞死亡。
Male hypogonadism (hgn/hgn) rats show testicular hypoplasia accompanied by dysplastic development of seminiferous tubules due to loss-of-function mutation of the gene encoding Astrin, which is required for mitotic progression in the division cycle of HeLa cells. In the present study, we examined the cytological base leading to the decrease of Sertoli cells inhgn/hgntestes. Inhgn/hgntestes on postnatal day 3, anti-phospho-histone H3 (Ser10) (pH3)-positive mitotic phase and TUNEL-positive apoptosis increased in GATA4-positive Sertoli cells. Isolated immature Sertoli cells fromhgn/hgntestes showed increased pH3-assessed mitotic index accompanied by decreased 5-bromo-2'-deoxyuridine-incorporation and increased TUNEL-positive apoptosis, suggesting mitotic delay and cell death. In the visualization of mitotic progression by nocodazole (NOC)-mediated cell cycle arrest and subsequent release,hgn/hgnrat-derived Sertoli cells failed to make the transition from prometaphase to metaphase, and the cells with micronuclei and TUNEL-positive cells gradually increased in a time-dependent manner. Western blot analysis detected ≈142 kDa protein expected as Astrin in extracts of +/+ and +/hgntestes and cultured normal Sertoli cells but not in extracts ofhgn/hgntestes. CLASP1 was detected in extracts of both normal andhgn/hgntestes, whereas it was localized in kinetochore of normal mitotic Sertoli cells but diffused in cytoplasm ofhgn/hgnSertoli cells. These results indicate that Astrin is required for normal mitotic progression in immature Sertoli cells and that the most severe type of testicullar dysplasia inhgn/hgnrats is caused by mitotic cell death of immature Sertoli cells due to lack of Astrin.