Barone Fortunata, Aguanno Salvatore, D'Alessio Alessio, D'Agostino Angela
Department of Histology and Medical Embryology, University of Rome "La Sapienza," Via A. Scarpa 14, 00161 Rome, Italy.
FASEB J. 2004 Feb;18(2):353-4. doi: 10.1096/fj.03-0347fje. Epub 2003 Dec 4.
Spontaneous cell death by apoptosis--occurring during normal spermatogenesis in mammals--is a prominent event, which results in the loss of up to 75% of the potential number of mature spermatozoa. In the rat testis, the most conspicuous dying cells are pachytene spermatocytes, which are also the primary target of the apoptosis experimentally induced by methoxyacetic acid (MAA). In this paper, we have used clusterin expression as an indicator of germ cell apoptosis in rat seminiferous tubules treated with MAA in the presence or in the absence of voltage-operated calcium channels (VOCCs) inhibitors. We performed both a qualitative analysis of clusterin expression by immunofluorescence experiments and a quantitative analysis of apoptosis by in situ end labeling of apoptotic germ cells followed by flow cytometry. The results obtained demonstrate that Sertoli cells, the somatic component of the seminiferous epithelium, which control male germ cell differentiation, also modulate MAA-induced apoptosis of germ cells throughout voltage-operated calcium channels.
在哺乳动物正常精子发生过程中发生的细胞凋亡导致的自发性细胞死亡是一个显著事件,其导致高达75%的潜在成熟精子数量损失。在大鼠睾丸中,最明显的死亡细胞是粗线期精母细胞,它们也是甲氧基乙酸(MAA)实验诱导凋亡的主要靶点。在本文中,我们使用簇集素表达作为在存在或不存在电压门控钙通道(VOCCs)抑制剂的情况下用MAA处理的大鼠生精小管中生殖细胞凋亡的指标。我们通过免疫荧光实验对簇集素表达进行了定性分析,并通过对凋亡生殖细胞进行原位末端标记后用流式细胞术对凋亡进行了定量分析。获得的结果表明,支持细胞是生精上皮的体细胞成分,其控制雄性生殖细胞分化,同时也通过电压门控钙通道调节MAA诱导的生殖细胞凋亡。