Expression of toxin-related human mono-ADP-ribosyltransferase 3 in human testes

Aim： To investigate wether the corresponding protein of mono-ADP-ribosyltransferase 3 （ART3） mRNA is expressed in human testes and, if so, whether the expression is cell type-specific. Methods： ART3 mRNA was determined in human testes and sperm by reverse transcription-polymerase chain reaction （RT-PCR）. The glycosylphosphatidylinositol linkage of ART3 was shown by treating ART3-transfected HEK-293-T cells with phospholipase C. Fluorescent activated cell sorter （FACS）-analyses were used to detect ART3 on mature spermatozoa and immunohistological studies to detect the protein in testes. Results： ART3 protein was shown to be present in testes. It was found on spermatocytes only. It was absent from spermatogonia, spermatids and spermatozoa. The absence of ART3 from spermatozoa was confirmed by FACS-analysis. ART3 protein was detected neither within a seminoma nor on Leydig cells. Conclusion： Here we show for the first time that ART3 protein is expressed in testes in particular on spermatocytes, indicating that ART3 exerts a specific function only required at a particular stage of spermatogenesis.

与毒素有关的人类单腺苷二磷酸-核糖基转移酶3(ART3)在人类睾丸中的表达

目的:研究单腺苷二磷酸-核糖基转移酶3(ART3)的 mRNA 在人类睾丸中是否表达其相应的蛋白,如果有表达是否细胞特异性表达。方法:用逆转录聚合酶链反应(RT-PCR)确定人类睾丸和精子中 ART3的mRNA 水平。用磷脂酶 C 处理转染了 ART3的 HEK-293-T 细胞,并通过它检测 ART3的糖基化磷酸肌醇链。用荧光激活细胞分类(FACS)分析和免疫组织学方法分别检测 ART3蛋白在成熟精子和睾丸中的表达。结果:ART3蛋白在睾丸和精母细胞中有表达,而在精原细胞、精子细胞和精子中未检测到,FACS 分析进一步确认精子中无 ART3蛋白。在精原细胞瘤和睾丸间细胞中也未检测到 ART3蛋白。结论:本研究首次发现ART3蛋白在睾丸中表达,尤其是在精母细胞中,这表明 ART3执行一个特定的功能,此功能只在精子发生的某个特殊阶段发挥作用。