Department of Physiological Sciences, Center for Environmental and Human Toxicology, University of Florida, Gainesville, FL 32611, USA.
Gen Comp Endocrinol. 2011 Aug 1;173(1):86-95. doi: 10.1016/j.ygcen.2011.04.027. Epub 2011 May 10.
Cholesterol transport across the mitochondrial membrane is rate-limiting for steroidogenesis in vertebrates. Previous studies in fish have characterized expression of the steroidogenic acute regulatory protein, however the function and regulation of other genes and proteins involved in piscine cholesterol transport have not been evaluated. In the current study, mRNA sequences of the 18 kDa translocator protein (tspo; formerly peripheral benzodiazepine receptor), voltage-dependent anion channel (vdac), and diazepam binding inhibitor (dbi; also acyl-CoA binding protein) were cloned from largemouth bass. Gonadal expression was examined across reproductive stages to determine if expression is correlated with changes in steroid levels and with indicators of reproductive maturation. In testis, transcript abundance of tspo and dbi increased with reproductive maturation (6- and 23-fold maximal increase, respectively) and expression of tspo and dbi was positively correlated with reproductive stage, gonadosomatic index (GSI), and circulating levels of testosterone. Testis vdac expression was positively correlated with reproductive stage and GSI. In females, gonadal tspo and vdac expression was negatively correlated with GSI and levels of plasma testosterone and 17β-estradiol. Ovarian dbi expression was not correlated with indicators of reproductive maturation. These studies represent the first investigation of the steroidogenic role of tspo, vdac, and dbi in fish. Findings suggest that cholesterol transport in largemouth bass testis, but not in ovary, may be transcriptionally-regulated, however further investigation will be necessary to fully elucidate the role of these genes in largemouth bass steroidogenesis.
胆固醇跨线粒体膜的转运是脊椎动物类固醇生成的限速步骤。鱼类的类固醇急性调节蛋白表达特征已有研究,然而,鱼类胆固醇转运中涉及的其他基因和蛋白质的功能和调节尚未得到评估。在本研究中,从大口黑鲈中克隆了 18kDa 转位蛋白(tspo;以前称为外周苯二氮䓬受体)、电压依赖性阴离子通道(vdac)和地西泮结合抑制剂(dbi;也称为酰基辅酶 A 结合蛋白)的 mRNA 序列。检测了性腺在繁殖阶段的表达,以确定其表达是否与类固醇水平的变化以及与生殖成熟的指标相关。在睾丸中,tspo 和 dbi 的转录丰度随着生殖成熟而增加(最大增加分别为 6 倍和 23 倍),并且 tspo 和 dbi 的表达与生殖阶段、性腺指数(GSI)和循环睾酮水平呈正相关。睾丸 vdac 的表达与生殖阶段和 GSI 呈正相关。在雌性中,性腺 tspo 和 vdac 的表达与 GSI 和血浆睾酮和 17β-雌二醇水平呈负相关。卵巢 dbi 的表达与生殖成熟的指标无关。这些研究代表了 tspo、vdac 和 dbi 在鱼类中的类固醇生成作用的首次研究。研究结果表明,大口黑鲈睾丸中的胆固醇转运可能受转录调控,但卵巢中的胆固醇转运可能不受转录调控,然而,需要进一步研究才能充分阐明这些基因在大口黑鲈类固醇生成中的作用。