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水通道蛋白8在大鼠睾丸中的表达与定位

Expression and localization of the aquaporin-8 water channel in rat testis.

作者信息

Calamita G, Mazzone A, Cho Y S, Valenti G, Svelto M

机构信息

Dipartimento di Fisiologia Generale ed Ambientale, Università degli Studi di Bari, 70126 Bari, Italy.

出版信息

Biol Reprod. 2001 Jun;64(6):1660-6. doi: 10.1095/biolreprod64.6.1660.

Abstract

Spermatogenesis and sperm maturation and storage are accompanied by significant movements of water, and multiple aquaporin transmembrane water channels (AQPs) have been recognized in the male reproductive tract. Nevertheless, the involvement of aquaporins in male reproductive physiology is mostly unknown. Here the expression and localization of AQP8 in rat spermatogenesis is defined and compared to that of AQP7, another aquaporin expressed in male germ cells. AQP8 mRNA was found in testis but not in epididymis, whereas the AQP7 transcript was present in both locations. By immunoblotting, the AQP8 protein was detected as a 25-kDa band and a 32- to 40-kDa diffuse component corresponding to the core and glycosylated protein, respectively. Membrane fractionation revealed AQP8 both in microsomal and plasma membrane-enriched fractions of rat testis while no apparent bands were detected in epididymis. AQP7 appeared as a 23- to 24-kDa band and was found both in testis and epididymis. By immunofluorescence, AQP8 labeling was found intracellularly as well as over the plasma membrane of germ cells throughout spermatogenesis. AQP7 was present in spermatids and spermatozoa and was predominant over the plasma membrane. AQP8 may be involved in the cytoplasmic condensation occurring during differentiation of spermatids into spermatozoa and in the generation of seminiferous tubule fluid.

摘要

精子发生、精子成熟和储存都伴随着显著的水的移动,并且在雄性生殖道中已识别出多种水通道蛋白跨膜水通道(AQP)。然而,水通道蛋白在雄性生殖生理学中的作用大多未知。在此,确定了AQP8在大鼠精子发生过程中的表达和定位,并将其与另一种在雄性生殖细胞中表达的水通道蛋白AQP7进行比较。在睾丸中发现了AQP8 mRNA,但在附睾中未发现,而AQP7转录本在这两个部位均有存在。通过免疫印迹法,检测到AQP8蛋白为一条25 kDa的条带以及一条32至40 kDa的弥散成分,分别对应核心蛋白和糖基化蛋白。膜分离显示,AQP8在大鼠睾丸的微粒体和富含质膜的部分中均有存在,而在附睾中未检测到明显条带。AQP7表现为一条23至24 kDa的条带,在睾丸和附睾中均有发现。通过免疫荧光法,在整个精子发生过程中,在生殖细胞的细胞内以及质膜上均发现了AQP8标记。AQP7存在于精子细胞和精子中,并且在质膜上占主导地位。AQP8可能参与精子细胞分化为精子过程中发生的细胞质浓缩以及生精小管液的生成。

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