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日本鹌鹑、日本草蜥和牛蛙松果体复合体中的视色素:免疫细胞化学和高效液相色谱分析

Visual pigments in the pineal complex of the Japanese quail, Japanese grass lizard and bullfrog: immunocytochemistry and HPLC analysis.

作者信息

Masuda H, Oishi T, Ohtani M, Michinomae M, Fukada Y, Shichida Y, Yoshizawa T

机构信息

Department of Biology, Faculty Science, Nara Women's University, Japan.

出版信息

Tissue Cell. 1994 Feb;26(1):101-13. doi: 10.1016/0040-8166(94)90086-8.

Abstract

We investigated localization of visual pigments in the pineal complex of Japanese quail, Japanese grass lizards and bullfrogs immunocytochemically by use of the antiserum against bovine rhodopsin (Rh-As) and monoclonal antibodies against chicken iodopsin (Io-mAb). We also analyzed retinoids, chromophores of visual pigments, by a high performance liquid chromatography (HPLC). The outer segments and cell membranes of some photoreceptor cells in the pineal organ of the Japanese quail exhibited immunoreactivity to Rh-As, but there are also many immunonegative cells. The number of immunoreactive cells among individuals varied. Immunoreactivity to Io-mAb was weak or did not exist. The HPLC analysis revealed peaks of 11-cis and all-trans isomers of retinal in the oxime extracts of the pineal organ of Japanese quail and chickens. In the pineal of Japanese grass lizards, the outer segments of some cells were immunopositive to Io-mAb, but there were no cells immunoreactive to Rh-As. The parietal eye exhibited a well-developed lens and photoreceptor cells, but the outer segments of photoreceptor cells were immunonegative to both Rh-As and Io-mAb. In bullfrogs, three types of cells were identified in both the pineal and frontal organ; (1) immunopositive to Rh-As, (2) immunopositive to Io-mAb and (3) immunonegative to either of the antibodies. In the pineal organ of bullfrogs, 11-cis and all-trans retinal and 11-cis 3-dehydroretinal were detected, and 11-cis and all-trans retinal were also detected in the frontal organ. We detected 11-cis and all-trans retinal in the ventral part of diencephalon including the hypothalamus. Thus, the chromophore is the same between the retinal and pineal visual pigments, but the expression of opsins is different between the retina and pineal complex, which probably reflects the different function of each organ.

摘要

我们通过使用抗牛视紫红质抗血清(Rh-As)和抗鸡视锥色素单克隆抗体(Io-mAb),采用免疫细胞化学方法研究了日本鹌鹑、日本草蜥和牛蛙松果体复合体中视觉色素的定位。我们还通过高效液相色谱法(HPLC)分析了作为视觉色素发色团的类视黄醛。日本鹌鹑松果体器官中一些光感受器细胞的外段和细胞膜对Rh-As呈现免疫反应性,但也有许多免疫阴性细胞。个体间免疫反应性细胞的数量各不相同。对Io-mAb的免疫反应性较弱或不存在。HPLC分析显示,日本鹌鹑和鸡松果体器官的肟提取物中存在视黄醛的11-顺式和全反式异构体峰。在日本草蜥的松果体中,一些细胞的外段对Io-mAb呈免疫阳性,但没有细胞对Rh-As呈免疫反应性。顶眼有发育良好的晶状体和光感受器细胞,但光感受器细胞的外段对Rh-As和Io-mAb均呈免疫阴性。在牛蛙中,松果体和额器官中均鉴定出三种类型的细胞:(1)对Rh-As呈免疫阳性,(2)对Io-mAb呈免疫阳性,(3)对任何一种抗体均呈免疫阴性。在牛蛙的松果体器官中,检测到了11-顺式和全反式视黄醛以及11-顺式3-脱氢视黄醛,在额器官中也检测到了11-顺式和全反式视黄醛。我们在下丘脑等间脑腹侧部分检测到了11-顺式和全反式视黄醛。因此,视网膜和松果体视觉色素的发色团相同,但视蛋白在视网膜和松果体复合体中的表达不同,这可能反映了每个器官的不同功能。

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