Kato Mutsuko, Sugiyama Takashi, Sakai Kazumi, Yamashita Takahiro, Fujita Hirofumi, Sato Keita, Tomonari Sayuri, Shichida Yoshinori, Ohuchi Hideyo
Department of Cytology and Histology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.
R&D Group, Olympus Corporation, Tokyo, Japan.
PLoS One. 2016 Nov 18;11(11):e0163925. doi: 10.1371/journal.pone.0163925. eCollection 2016.
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.
视蛋白家族基因编码G蛋白偶联的七跨膜蛋白,这些蛋白在光感受过程中结合视黄醛发色团。在此,我们寻找潜在的尚未被描述的鸟类视网膜光感受器,重点关注鸡的视蛋白3同源物。我们在鸡基因组中发现了两个与视蛋白3相关的基因:一个对应于哺乳动物中的脑视蛋白/泛视蛋白(Opn3),另一个属于硬骨鱼多组织视蛋白(TMT)2组。生物发光成像和G蛋白激活试验表明,鸡TMT视蛋白(cTMT)在哺乳动物培养细胞中强制表达时可作为蓝光传感器发挥作用。我们没有检测到鸡Opn3(cOpn3)对光敏感的证据。原位杂交显示cTMT在内视网膜分化细胞亚群中表达,并且随着发育进程,主要定位于视网膜水平细胞(HCs)。免疫组织化学(IHC)显示cTMT存在于HCs以及孵化后鸡视网膜神经节和内核层中的少量细胞中。相比之下,cOpn3免疫反应阳性(IR)细胞在内核层的不同细胞亚群中被发现。在下丘脑的细胞亚群中也发现了cTMT-IR细胞。最后,我们发现cOpn3和cTMT蛋白在小脑的特定细胞中有差异分布。目前的结果表明,一种新型的TMT型视蛋白3可能在鸡的视网膜和大脑中作为光感受器发挥作用。