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胆固醇转运蛋白ABCA1和ABCG1在犬生殖道组织和精子中的免疫定位

Immunolocalization of the cholesterol transporters ABCA1 and ABCG1 in canine reproductive tract tissues and spermatozoa.

作者信息

Palme N, Becher A C, Merkl M, Glösmann M, Aurich C, Schäfer-Somi S

机构信息

Platform for Artificial Insemination and Embryo Transfer, Vetmeduni Vienna, Vienna, Austria.

出版信息

Reprod Domest Anim. 2014 Jun;49(3):441-7. doi: 10.1111/rda.12294. Epub 2014 Mar 11.

Abstract

The mammalian sperm membrane undergoes cholesterol efflux during maturation and fertilization. Although ATP-binding cassette (ABC) transporters are known to transport cholesterol through cell membranes in other organs, their presence in canine testis, epididymis and sperm has not been proven to date. Hence, the aim of the present study was to localize the ABC transporters ABCA1 and ABCG1 in canine testicular and epididymidal tissue as well as in spermatozoa membranes. To this end, semen samples from 12 dogs as well as testicles and epididymides of four young and healthy dogs were prepared for immunohistochemistry, respectively. Capacitation and acrosome reaction (AR) were induced in aliquots of the semen samples before immunostaining to assess changes in the expression of ABCA1 and ABCG1. Evaluation by confocal microscopy revealed the presence of both ABCA1 and ABCG1 in canine testicles and of ABCA1 in the epididymides. In spermatozoa, only ABCA1 immunoreactivity was detected, mainly in the region of the acrosome and midpiece. After induction of capacitation, ABCA1 signal persisted in the acrosome but disappeared after AR, indicating a loss of ABCA1 with the loss of the acrosome. We conclude that ABCA1 and ABCG1 are expressed in canine testis, whereas only ABCA1 is expressed in epididymis and spermatozoa membrane, both transporters probably contributing to the regulation of membrane cholesterol content.

摘要

哺乳动物精子膜在成熟和受精过程中会发生胆固醇外流。尽管已知ATP结合盒(ABC)转运蛋白可在其他器官中通过细胞膜转运胆固醇,但迄今为止,它们在犬类睾丸、附睾和精子中的存在尚未得到证实。因此,本研究的目的是在犬类睾丸和附睾组织以及精子细胞膜中定位ABC转运蛋白ABCA1和ABCG1。为此,分别对12只犬的精液样本以及4只年轻健康犬的睾丸和附睾进行免疫组织化学制备。在免疫染色前,对精液样本的等分试样进行获能和顶体反应(AR)诱导,以评估ABCA1和ABCG1表达的变化。通过共聚焦显微镜评估发现,ABCA1和ABCG1均存在于犬类睾丸中,ABCA1存在于附睾中。在精子中,仅检测到ABCA1免疫反应性,主要位于顶体和中段区域。获能诱导后,ABCA1信号在顶体中持续存在,但在AR后消失,表明随着顶体的丧失,ABCA1也丧失。我们得出结论,ABCA1和ABCG1在犬类睾丸中表达,而仅ABCA1在附睾和精子细胞膜中表达,这两种转运蛋白可能都有助于调节膜胆固醇含量。

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