Abstract

Translocator protein (TSPO) present in the outer mitochondrial membrane has been suggested to be critical for cholesterol import, a rate-limiting step for steroid hormone biosynthesis. Despite the importance of steroidogenesis in regulating reproductive functions, the developmental profile of TSPO expression in the gonads and accessory sex organs has not been completely characterized. As a first step towards understanding the function of TSPO, we studied its expression in male and female murine reproductive organs. We examined testes and ovaries at embryonic days 14.5 and 18.5, and postnatal days 0, 7, 14, 21 and 56 of development. In the adult testis, TSPO was expressed in both Leydig cells and Sertoli cells. In the developing testes TSPO expression was seen in immature Sertoli cells, fetal Leydig cells and gonocytes. In the ovary, TSPO was expressed in the ovarian surface epithelium, interstitial cells granulosa cells and luteal cells. Corpora lutea of ovaries from pregnant mice showed strong expression of TSPO. In the developing ovary, TSPO expression was seen in the squamous pregranulosa cells associated with germ line cysts, together with progressively increasing expression in interstitial cells and the ovarian surface epithelium. In adult mice, the epithelia of other reproductive tissues like the epididymis, prostate, seminal vesicle, oviduct and uterus also showed distinct patterns of TSPO expression. In summary, TSPO expression in both male and female reproductive tissues was not only restricted to steroidogenic cells. Expression in Sertoli cells, ovarian surface epithelium, efferent ductal epithelium, prostatic epithelium, seminal vesiclular epithelium, uterine epithelium and oviductal epithelium suggest either previously unknown sites for de novo steroidogenesis or functions for TSPO distinct from its well-studied role in steroid hormone production.

Highlights

  • Translocator protein (TSPO) was first identified as a pharmacologically distinct diazepam-binding protein [1,2], and has long been studied under its former name, peripheral-type benzodiazepine receptor (PBR) [3]

  • In western blots from protein lysates from a Leydig cell line (MA-10), the rabbit monoclonal TSPO antibody recognized a single 18 kDa band that corresponds to the expected molecular weight defined by the murine TSPO cDNA sequence

  • TSPO fluorescence in MA-10 cells colocalized with the mitochondrial marker VDAC1 (Fig. 2)

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Summary

Introduction

Translocator protein (TSPO) was first identified as a pharmacologically distinct diazepam-binding protein [1,2], and has long been studied under its former name, peripheral-type benzodiazepine receptor (PBR) [3]. Biochemical characterization of this 18 kDa transmembrane protein showed predominant presence in the mitochondria, with specific localization to the mitochondrial outer membrane [4,5]. TSPO gene expression is regulated by several mechanisms in tissues and is not completely defined. A mechanism of regulation of TSPO expression by a natural antisense transcript called short interspersed repetitive element B2 (SINE B2) has been shown to regulate TSPO transcription [29]

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