Schmitt M C, Ong D E
Department of Biochemistry, Vanderbilt University, Nashville, Tennessee 37232.
Biol Reprod. 1993 Nov;49(5):972-9. doi: 10.1095/biolreprod49.5.972.
Vitamin A deficiency in mammals results in the loss of germ cells on the adluminal side of the blood-testis barrier, suggesting a need for vitamin A that would be supplied by the surrounding Sertoli cells. Cellular retinol-binding protein (CRBP) and lecithin-retinol acyltransferase (LRAT) are two proteins found in Sertoli cells that are known to be involved in vitamin A trafficking. To clarify the role of these two proteins in the delivery of vitamin A to developing germ cells, we have examined changes in their cell-specific expression during the onset of puberty in the rat. In adult rats, Sertoli cell expression of CRBP varies with the cycle of the seminiferous epithelium. Here, we demonstrate that differences in the intensity of CRBP immunoreactivity are detectable in Sertoli cells of different tubules as early as postnatal Day 4, prior to the onset of meiosis. This indicates that variable expression of CRBP by Sertoli cells is established independently of late germ cells and may anticipate the cyclical variation seen in the adult. We further demonstrate that the specific activity of LRAT in rat testis increases tenfold between postnatal Days 20 and 35. This increase is attributable to the appearance of post-meiotic germ cells: the LRAT activity of microsomes prepared from a round spermatid-enriched cell fraction from post-pubertal rat testis could account for the majority of the LRAT activity observed in the whole testis. The presence of LRAT activity within adluminal germ cells suggests that they receive vitamin A as retinol and synthesize the retinyl esters that have been shown to be present in mature sperm.
哺乳动物体内维生素A缺乏会导致血睾屏障管腔侧生殖细胞的丢失,这表明发育中的生殖细胞需要周围支持细胞提供维生素A。细胞视黄醇结合蛋白(CRBP)和卵磷脂-视黄醇酰基转移酶(LRAT)是在支持细胞中发现的两种蛋白,已知它们参与维生素A的转运。为了阐明这两种蛋白在向发育中的生殖细胞输送维生素A中的作用,我们研究了它们在大鼠青春期开始期间细胞特异性表达的变化。在成年大鼠中,CRBP在支持细胞中的表达随生精上皮的周期而变化。在此,我们证明早在出生后第4天,即在减数分裂开始之前,在不同曲细精管的支持细胞中就可检测到CRBP免疫反应强度的差异。这表明支持细胞中CRBP的可变表达独立于晚期生殖细胞而建立,并且可能预示着在成年期观察到的周期性变化。我们进一步证明,大鼠睾丸中LRAT的比活性在出生后第20天至35天之间增加了10倍。这种增加归因于减数分裂后生殖细胞的出现:从青春期后大鼠睾丸的富含圆形精子细胞的细胞组分制备的微粒体的LRAT活性可占整个睾丸中观察到的LRAT活性的大部分。管腔侧生殖细胞中存在LRAT活性表明它们以视黄醇形式接受维生素A并合成已证明存在于成熟精子中的视黄酯。