Nakayama T, Furuya S
National Institute for Physiological Sciences, Okazaki, Japan.
Brain Res Dev Brain Res. 1991 Sep 19;62(1):99-108. doi: 10.1016/0165-3806(91)90194-n.
Differentiation of photoreceptor cells dissociated from chick embryo was investigated in a monolayer culture at low cell density and low serum concentration. The precursor cells were initially round and finally adopted the elongated and monopolar shape characteristic of such photoreceptor cells in vivo. They exhibited the primitive inner segment, and on its distal side showed a crest-like membrane expansion instead of the developed outer segment. The cell polarity of these cultured photoreceptor cells was examined using mushroom lectin, which has specific affinity for the D-galactose-beta(1,3)-N-acetylgalactosamine moiety. Rhodamine-labeled mushroom lectin uniformly initially stained the surface of early round photoreceptor cells, but with time the staining became restricted to the inner segment and its distal membrane expansion in all elongated photoreceptor cells at 12 days from the beginning of development. In semithin sections of 12-day-old chick embryo retina, mushroom lectin also stained the immature inner segment that had emerged at this stage. After hatching, light and electron microscopic observations revealed that the entire scleral surface of retinal plasma membranes as far as the outer limiting membrane, i.e. the inner and outer segments of all types of photoreceptor cells and also Müller cell processes, were stained with the mushroom lectin. These results clearly show that all photoreceptor cells can establish their inherent cell polarity even in culture, and that mushroom lectin is a valuable marker, for the investigation of photoreceptor cell polarity.
在低细胞密度和低血清浓度的单层培养中，研究了从鸡胚中分离出的光感受器细胞的分化情况。前体细胞最初呈圆形，最终呈现出体内此类光感受器细胞特有的细长单极形状。它们表现出原始的内段，并且在其远端一侧显示出嵴状的膜扩张，而不是发育成熟的外段。使用对D-半乳糖-β(1,3)-N-乙酰半乳糖胺部分具有特异性亲和力的蘑菇凝集素，检测这些培养的光感受器细胞的细胞极性。罗丹明标记的蘑菇凝集素最初均匀地染色早期圆形光感受器细胞的表面，但随着时间的推移，在发育开始12天后，所有细长的光感受器细胞中的染色都局限于内段及其远端的膜扩张。在12日龄鸡胚视网膜的半薄切片中，蘑菇凝集素也对此时出现的未成熟内段进行了染色。孵化后，光镜和电镜观察显示，视网膜质膜的整个巩膜表面直至外限制膜，即所有类型光感受器细胞的内段和外段以及米勒细胞突起，都被蘑菇凝集素染色。这些结果清楚地表明，即使在培养中，所有光感受器细胞也能建立其固有的细胞极性，并且蘑菇凝集素是研究光感受器细胞极性的一种有价值的标记物。
