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Two Opsin 3-Related Proteins in the Chicken Retina and Brain: A TMT-Type Opsin 3 Is a Blue-Light Sensor in Retinal Horizontal Cells Hypothalamus and Cerebellum

机译:鸡视网膜和大脑中两种与视蛋白3相关的蛋白:TMT型视蛋白3是视网膜水平细胞下丘脑和小脑中的蓝光传感器。

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摘要

Opsin family genes encode G protein-coupled seven-transmembrane proteins that bind a retinaldehyde chromophore in photoreception. Here, we sought potential as yet undescribed avian retinal photoreceptors, focusing on Opsin 3 homologs in the chicken. We found two Opsin 3-related genes in the chicken genome: one corresponding to encephalopsin/panopsin (Opn3) in mammals, and the other belonging to the teleost multiple tissue opsin (TMT) 2 group. Bioluminescence imaging and G protein activation assays demonstrated that the chicken TMT opsin (cTMT) functions as a blue light sensor when forced-expressed in mammalian cultured cells. We did not detect evidence of light sensitivity for the chicken Opn3 (cOpn3). In situ hybridization demonstrated expression of cTMT in subsets of differentiating cells in the inner retina and, as development progressed, predominant localization to retinal horizontal cells (HCs). Immunohistochemistry (IHC) revealed cTMT in HCs as well as in small numbers of cells in the ganglion and inner nuclear layers of the post-hatch chicken retina. In contrast, cOpn3-IR cells were found in distinct subsets of cells in the inner nuclear layer. cTMT-IR cells were also found in subsets of cells in the hypothalamus. Finally, we found differential distribution of cOpn3 and cTMT proteins in specific cells of the cerebellum. The present results suggest that a novel TMT-type opsin 3 may function as a photoreceptor in the chicken retina and brain.
机译:视蛋白家族基因编码G蛋白偶联的七跨膜蛋白,它们在光接收中结合视黄醛发色团。在这里,我们寻求潜在的尚未描述的鸟类视网膜感光器,重点研究鸡中的Opsin 3同源物。我们在鸡基因组中发现了两个与Opsin 3相关的基因:一个对应于哺乳动物的脑视蛋白/ panopsin(Opn3),另一个属于硬骨多组织视蛋白(TMT)2组。生物发光成像和G蛋白活化测定表明,鸡TMT视蛋白(cTMT)在哺乳动物培养细胞中强制表达时起蓝光传感器的作用。我们没有发现鸡Opn3(cOpn3)有光敏感性的证据。原位杂交证明了cTMT在内部视网膜分化细胞亚群中的表达,并且随着发育的发展,主要定位于视网膜水平细胞(HCs)。免疫组织化学(IHC)揭示了HCs中以及孵化后鸡视网膜神经节和内核层中少量细胞中的cTMT。相反,在内部核层的不同细胞亚群中发现了cOpn3-IR细胞。在下丘脑的细胞亚群中也发现了cTMT-IR细胞。最后,我们在小脑的特定细胞中发现了cOpn3和cTMT蛋白的差异分布。目前的结果表明,新型的TMT型视蛋白3可以在鸡的视网膜和大脑中充当感光器。

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