Bastianelli E, Takamatsu K, Okazaki K, Hidaka H, Pochet R
Laboratoires d'Anatomie Pathologique, Faculté de Médecine, Université Libre de Bruxelles, Belgium.
Exp Eye Res. 1995 Mar;60(3):257-66. doi: 10.1016/s0014-4835(05)80108-3.
The post-natal developmental expression in rat retina of four calcium-binding proteins belonging to the calmodulin-troponin-C family was investigated by immunohistochemistry using anti-calbindin-D28k, anti-calretinin, anti-hippocalcin and anti-neurocalcin polyclonal antibodies on paraffin sections from Wistar rat retinae aged from post-natal days 1 (P1), 5 (P5), 10 (P10), 20 (P20) to adulthood (8 weeks). Immunoblot using anti-hippocalcin and homogenates proteins from retina, cerebellar cortex, hippocampus and cerebellum was also performed. Hippocalcin immunoreactivity in adult rat retina was demonstrated by both immunohistochemistry and Western blot. During post-natal development, calbindin-D28k, calretinin and neurocalcin immunoreactivity were detected at P1 in ganglion cells, whereas hippocalcin immunoreactivity was seen later at P5 in this cell layer. In the amacrine cell layer, neurocalcin immunoreactivity was detected at P5 and hippocalcin at P10. Calbindin-D28k was labelling the immature horizontal cell, calretinin was detected in nearly all ganglion cells and in some amacrine cells since P1. These three calcium-binding proteins do not seem to play a role in synaptogenesis which takes place later. We confirmed that calbindin-D28k appeared to be a good marker for horizontal cells. The presence of hippocalcin, a myristoylated calcium-binding protein belonging to the recovering subfamily and previously localized in few brain areas has been detected for the first time in retina.
利用抗钙结合蛋白-D28k、抗钙视网膜蛋白、抗海马钙结合蛋白和抗神经钙蛋白多克隆抗体，通过免疫组织化学方法，对出生后1天（P1）、5天（P5）、10天（P10）、20天（P20）至成年期（8周）的Wistar大鼠视网膜石蜡切片进行研究，以探讨钙调蛋白-肌钙蛋白-C家族中四种钙结合蛋白在大鼠视网膜中的产后发育表达情况。同时，还使用抗海马钙结合蛋白对视网膜、小脑皮质、海马和小脑的匀浆蛋白进行了免疫印迹分析。免疫组织化学和蛋白质印迹法均证实成年大鼠视网膜中存在海马钙结合蛋白免疫反应性。在出生后的发育过程中，钙结合蛋白-D28k、钙视网膜蛋白和神经钙蛋白免疫反应性在P1时在神经节细胞中被检测到，而海马钙结合蛋白免疫反应性在该细胞层中较晚在P5时出现。在内网层细胞层中，神经钙蛋白免疫反应性在P5时被检测到，海马钙结合蛋白在P10时被检测到。钙结合蛋白-D28k标记未成熟的水平细胞，自P1起在几乎所有神经节细胞和一些内网层细胞中检测到钙视网膜蛋白。这三种钙结合蛋白似乎在后期发生的突触形成过程中不起作用。我们证实钙结合蛋白-D28k似乎是水平细胞的良好标记物。首次在视网膜中检测到海马钙结合蛋白，它是一种属于恢复亚家族的肉豆蔻酰化钙结合蛋白，以前仅在少数脑区定位。
