Halfter W, Fua C S
Max-Planck Guest Laboratory, Institute of Cell Biology, Academia Sinica, Shanghai, China.
Cell Tissue Res. 1987 Sep;249(3):487-96. doi: 10.1007/BF00217320.
Antibodies against laminin (LN), fibronectin (FN), collagen type IV (Col IV), neural cell adhesion molecule (N-CAM), T-61 antigen, actin, tubulin and neurofilament protein were injected into the eyes of quail embryos (Coturnix coturnix japonica) of different ages. Twenty h after injection, the heads of the embryos were fixed and the antibodies visualized in sections with the use of fluorescein-isothiocyanate (FITC) or peroxidase-labeled second antibodies by light- and electron microscopy. Antibodies against cell surface molecules, such as N-CAM, LN, Col IV and T 61, labeled matrix and membrane components of the retinal cells in different antigen-specific patterns. Antibodies against intracellular antigens, such as actin, tubulin and neurofilament protein labeled nonspecifically the vitreous body and the inner basal lamina of the retina, but resulted in only a very weak and diffuse labeling of retinal cells. N-CAM was detected in high concentration in the optic fiber layer on the surface of axons and on the membranes of all retinal cells. Col IV, LN and T 61 antigen were found predominantly in the optic fiber layer. LN and Col IV were located on the surface of axons and the endfeet of ventricular (neuroepithelial) cells in a patchy distribution. The T-61 antigen was found in early stages in the cell-free space of the optic fiber layer, on the surface of ventricular cells and axons, and at later stages also in high-density patches between nerve fibers. The distribution of LN and T-61 antigen together with data from in vitro experiments suggests a crucial role of these proteins in axon extension in the avian retina during early development of the optic fiber layer.
将抗层粘连蛋白(LN)、纤连蛋白(FN)、IV型胶原(Col IV)、神经细胞黏附分子(N-CAM)、T-61抗原、肌动蛋白、微管蛋白和神经丝蛋白的抗体注入不同年龄的鹌鹑胚胎(日本鹌鹑)眼中。注射后20小时,将胚胎头部固定,通过光学显微镜和电子显微镜,使用异硫氰酸荧光素(FITC)或过氧化物酶标记的二抗在切片中使抗体可视化。针对细胞表面分子(如N-CAM、LN、Col IV和T 61)的抗体,以不同的抗原特异性模式标记视网膜细胞的基质和膜成分。针对细胞内抗原(如肌动蛋白、微管蛋白和神经丝蛋白)的抗体非特异性地标记玻璃体和视网膜的内基底层,但仅导致视网膜细胞非常微弱和弥散的标记。在轴突表面的视神经纤维层和所有视网膜细胞的膜上检测到高浓度的N-CAM。Col IV、LN和T 61抗原主要存在于视神经纤维层。LN和Col IV以斑片状分布位于轴突表面和室管膜(神经上皮)细胞的终足上。T-61抗原在早期出现在视神经纤维层的无细胞空间、室管膜细胞和轴突表面,在后期也出现在神经纤维之间的高密度斑块中。LN和T-61抗原的分布以及体外实验数据表明,这些蛋白质在视神经纤维层早期发育过程中对鸟类视网膜轴突延伸起着关键作用。