Shim K, Picking W L, Kutty R K, Thomas C F, Wiggert B N, Stark W S
Department of Biology, Saint Louis University, St. Louis, MO 63103-2010, USA.
Exp Eye Res. 1997 Nov;65(5):717-27. doi: 10.1006/exer.1997.0383.
In Drosophila, thorough retinoid deprivation is possible, optimizing investigation of the effects of vitamin A metabolites and retinoic acid on the visual system. Retinoids had been found to control transcription and translation of Drosophila's opsin gene. To follow this line of inquiry, we examined the effect of retinoids on the translation and transcription of a Drosophila Retinoid and Fatty Acid Binding Glycoprotein. Western blots showed that this protein is high in retinoid replete flies and low in deprived flies. Flies grown on media capable of activating the opsin gene's transcription and which contain alternate transcription activators including retinoic acid yielded extracts containing significant amounts of Retinoid and Fatty Acid Binding Glycoprotein. Immunocytochemistry confirmed its absence in deprived flies and its presence in flies reared or replaced on these diverse media containing retinoids or general nutrients. Immunocytochemistry localized Retinoid and Fatty Acid Binding Glycoprotein to the Semper (cone) cells and the intraommatidial matrix (the interphotoreceptor matrix of the ommatidium). Positive staining of Semper cells in mutants of the opsin gene and a mutant lacking receptors suggests that Retinoid and Fatty Acid Binding Glycoprotein does not depend on presence of opsin and that it is not synthesized in receptor cells respectively. Northern blots demonstrated greatly diminished mRNA for Retinoid and Fatty Acid Binding Glycoprotein in flies grown on deprivation food relative to flies grown on normal food. Although the synthesis of Retinoid and Fatty Acid Binding Glycoprotein does not require chromophore precursors as does that of opsin, the control of Retinoid and Fatty Acid Binding Glycoprotein and opsin transcription by retinoids including retinoic acid might very well be the same. Our results suggest that Retinoid and Fatty Acid Binding Glycoprotein may be involved in retinoid transport. Also, Semper cells may be analogous to vertebrate retinal pigment epithelium in retinoid metabolism and/or delivery.
在果蝇中,可以实现彻底的类视黄醇剥夺,从而优化对维生素A代谢物和视黄酸对视觉系统影响的研究。已发现类视黄醇可控制果蝇视蛋白基因的转录和翻译。为了沿着这条研究思路进行探究,我们研究了类视黄醇对果蝇类视黄醇和脂肪酸结合糖蛋白翻译和转录的影响。蛋白质免疫印迹显示,这种蛋白质在富含类视黄醇的果蝇中含量很高,而在缺乏类视黄醇的果蝇中含量很低。在能够激活视蛋白基因转录且含有包括视黄酸在内的其他转录激活剂的培养基上生长的果蝇,其提取物中含有大量的类视黄醇和脂肪酸结合糖蛋白。免疫细胞化学证实,在缺乏类视黄醇的果蝇中没有这种蛋白,而在以这些含有类视黄醇或一般营养物质的不同培养基饲养或更换饲养条件的果蝇中存在这种蛋白。免疫细胞化学将类视黄醇和脂肪酸结合糖蛋白定位到森珀(视锥)细胞和小眼内基质(小眼的光感受器间基质)。视蛋白基因的突变体和缺乏受体的突变体中小眼细胞的阳性染色表明,类视黄醇和脂肪酸结合糖蛋白不依赖于视蛋白的存在,并且它不是分别在受体细胞中合成的。Northern印迹显示,相对于在正常食物上生长的果蝇,在缺乏类视黄醇食物上生长的果蝇中,类视黄醇和脂肪酸结合糖蛋白的mRNA大大减少。虽然类视黄醇和脂肪酸结合糖蛋白的合成不像视蛋白那样需要发色团前体,但包括视黄酸在内的类视黄醇对类视黄醇和脂肪酸结合糖蛋白以及视蛋白转录的控制很可能是相同的。我们的结果表明,类视黄醇和脂肪酸结合糖蛋白可能参与类视黄醇的运输。此外,在类视黄醇代谢和/或传递方面,森珀细胞可能类似于脊椎动物的视网膜色素上皮。