Dobi E T, Troccoli N M, Hausman R E
Invest Ophthalmol Vis Sci. 1986 Mar;27(3):323-9.
The histological distribution of R-cognin in chick retinas was determined from embryonic day 8 through 13 wk post-hatching by indirect immunofluorescence using polyclonal anticognin. On embryonic day 8, at a developmental stage without distinct retina layers, most of the cells within the tissue exhibited fluorescence. By embryonic day 12, when the strata of the retina are delineating and initial synapses are beginning to form, R-cognin fluorescence became concentrated in the nascent ganglion cell, and to a lesser extent, inner nuclear layers. By embryonic day 16 (extending through post-hatching day 26), the staining of R-cognin was specific for the ganglion cell and nerve fiber layers. Fluorescence was prominent on the cell somal membranes but also was present on the processes of these cells at the ganglion cell layer-inner plexiform layer interface. Pre-immune serum and anticognin after preabsorption with R-cognin exhibited no fluorescence. The results demonstrated that the known decrease in R-cognin found in the retina during the latter half of embryonic development in the chick is not uniform across the retina, but that R-cognin is preferentially retained on cells within the ganglion cell layer. While cells within the ganglion cell layer also exhibit alpha-bungarotoxin binding, the majority of the latter is found in the inner plexiform layer. Thus, the observations are consistent with a role for R-cognin in the formation or maintenance of functional cell-cell connections within the entire retina prior to developmental day 11, or in retinal ganglion cell layer formation or stability subsequent to embryonic day 11.
通过使用多克隆抗认知蛋白的间接免疫荧光法,确定了从孵化后第8天胚胎期到第13周雏鸡视网膜中R - 认知蛋白的组织学分布。在胚胎第8天,处于视网膜各层尚不明显的发育阶段时,组织内的大多数细胞呈现荧光。到胚胎第12天,当视网膜各层开始分化且初始突触开始形成时,R - 认知蛋白荧光集中在新生的神经节细胞中,在内核层中的荧光较弱。到胚胎第16天(持续到孵化后第26天),R - 认知蛋白的染色在神经节细胞层和神经纤维层中具有特异性。荧光在细胞体膜上很突出,但在神经节细胞层 - 内网层界面处这些细胞的突起上也存在。预免疫血清以及用R - 认知蛋白预吸收后的抗认知蛋白均未显示荧光。结果表明,在雏鸡胚胎发育后半期视网膜中发现的R - 认知蛋白的已知减少在整个视网膜中并不均匀,而是R - 认知蛋白优先保留在神经节细胞层内的细胞上。虽然神经节细胞层内的细胞也表现出α - 银环蛇毒素结合,但后者大部分存在于内网层中。因此,这些观察结果与R - 认知蛋白在胚胎发育第11天之前在整个视网膜内功能性细胞 - 细胞连接的形成或维持中起作用,或在胚胎第11天之后在视网膜神经节细胞层的形成或稳定性中起作用是一致的。