DISTRIBUTION PATTERN OF 3 NEURAL CALCIUM-BINDING PROTEINS (NCS-1, VILIP AND RECOVERIN) IN CHICKEN, BOVINE AND RAT RETINA
DISTRIBUTION PATTERN OF 3 NEURAL CALCIUM-BINDING PROTEINS (NCS-1, VILIP AND RECOVERIN) IN CHICKEN, BOVINE AND RAT RETINA
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DOI：
10.1007/bf00174325
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发表时间：
1995-07-01
期刊：
影响因子：
--
通讯作者：
COX, JA
中科院分区：
文献类型：
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作者：
DERAAD, S;COMTE, M;COX, JA
神经钙结合蛋白（Neural Ca 2 + binding proteins，NCaPs）是4-EF-手蛋白的一个亚家族，在组织学和结构上呈现二分性：A型NCaPs选择性地表达于视网膜和松果体，呈现两个典型的EF-手，而B型NCaPs存在于整个脑中，呈现三个规则的EF-手。在这项研究中，产生了针对A型NCaP恢复素（26 kDa）和B型NCaP VILIP和NCS-1（22 kDa）的抗血清。由于NCaPs之间的序列同一性高，特异性的多克隆抗体通过双交叉免疫亲和层析纯化; ELISA和免疫印迹分析确定，所得抗体显示出对其他两个相关NCaPs的选择性比低于1/363。此外，抗VILIP抗体对神经钙蛋白δ显示出一定的亲和力，抗恢复素抗体识别24 kDa的蛋白质，其最有可能是visinin。因此，鸡，大鼠和牛视网膜的免疫组织化学研究表明，抗recoverin抗体识别脊椎动物的光感受器和少数哺乳动物的双极细胞。抗VILIP抗体仅标记内层视网膜，即无长突细胞和神经节细胞。NCS-1主要表达于感光细胞内节、内网状层和神经节细胞。NCS-1显示出最高的物种差异。NCS-1和VILIP的视网膜定位为了解其功能提供了重要的形态学基础。此外，针对NCaP的特异性抗体可以鉴定更复杂的神经组织（如大脑）中的细胞群。
Neural Ca2+-binding proteins (NCaPs) constitute a subfamily of 4-EF-hand proteins, and display a histological and structural dichotomy: the A-type NCaPs are selectively expressed by the retina and pineal organ and display two canonical EF-hands, whereas the B-type NCaPs are found in the entire brain and present three regular EF-hands. In this study, antisera were raised against the A-type NCaP recoverin (26 kDa) and the B-type NCaPs VILIP and NCS-1 (22 kDa). Since the sequence identity among NCaPs is high, specific polyclonal antibodies were purified by double cross-immunoaffinity chromatography; both ELISA and immunoblot analyses determined that the resulting antibodies showed selectivity ratios inferior to 1/363 for the two other related NCaPs. Besides, the anti-VILIP antibodies displayed some affinity toward neurocalcin delta, and the anti-recoverin antibodies recognized a 24 kDa protein, which is most likely visinin. Thus, immunohistochemical studies on the chicken, rat and cow retina revealed that anti-recoverin antibodies recognized the vertebrate photoreceptors and a small number of mammalian bipolar cells. Anti-VILIP antibodies exclusively labelled the inner retina, i.e. the amacrine and ganglion cells. NCS-1 was mainly present in the photoreceptor inner segments, the inner plexiform layer and the ganglion cells. NCS-1 showed the highest species disparity. The retinal localization of NCS-1 and VILIP offered an important morphological basis for the understanding of their function. Furthermore, specific antibodies against the NCaPs may enable the identification of cell populations in more complex neural tissues, such as the brain.