Koji T, Hishikawa Y, Ando H, Nakanishi Y, Kobayashi N
Department of Histology and Cell Biology, Nagasaki University School of Medicine, Nagasaki 852-8523, Japan.
Biol Reprod. 2001 Mar;64(3):946-54. doi: 10.1095/biolreprod64.3.946.
Apoptosis of germ cells is very common in normal and injured mammalian testes. The aim of this study was to examine the possible involvement of the Fas and Fas ligand (FasL) system in the induction of germ cell apoptosis in normal and ischemia-reperfusion testes of adult mice. Apoptosis was assessed by the TUNEL method and by DNA gel electrophoresis. Fas and FasL mRNAs were detected by Northern blotting and reverse transcription polymerase chain reaction techniques, and proteins were analyzed by Western blotting and immunohistochemistry. Apoptosis of germ cells was identified in the normal testis especially around stages XI and XII, whereas the expression of Fas and FasL was largely confined to Leydig cells and Sertoli cells, respectively. However, in the testes reperfused after 1 h of ischemia, a high number of TUNEL-positive cells were identified in parallel with increased Fas-positive germ cells, whereas FasL expression in Sertoli cells was almost constant irrespective of the duration of reperfusion. Moreover, i.p. injection of anti-Fas antibody, which blocks the interaction between Fas and FasL, inhibited apoptosis, as indicated by the reduced number of TUNEL-positive cells, except for apoptosis at stages XI and XII. Our results indicate that the Fas/FasL system mediates apoptosis of spermatogenic cells in the injured testis but not spontaneous apoptosis in the normal testis.
生殖细胞凋亡在正常及受损的哺乳动物睾丸中非常普遍。本研究旨在探讨Fas和Fas配体(FasL)系统在成年小鼠正常及缺血再灌注睾丸中诱导生殖细胞凋亡过程中可能发挥的作用。通过TUNEL法和DNA凝胶电泳评估细胞凋亡情况。采用Northern印迹法和逆转录聚合酶链反应技术检测Fas和FasL mRNA,并用蛋白质印迹法和免疫组织化学法分析蛋白质。在正常睾丸中,尤其是在第XI和XII期左右可识别出生殖细胞凋亡,而Fas和FasL的表达分别主要局限于睾丸间质细胞和支持细胞。然而,在缺血1小时后再灌注的睾丸中,可识别出大量TUNEL阳性细胞,同时Fas阳性生殖细胞数量增加,而无论再灌注时间长短,支持细胞中FasL的表达几乎恒定。此外,腹腔注射抗Fas抗体可阻断Fas与FasL之间的相互作用,除了第XI和XII期的凋亡外,TUNEL阳性细胞数量减少表明其抑制了细胞凋亡。我们的结果表明,Fas/FasL系统介导受损睾丸中生精细胞的凋亡,但不介导正常睾丸中的自发凋亡。