Wolburg H, Reichelt W, Stolzenburg J U, Richter W, Reichenbach A
Institute of Pathology, University of Tübingen, F.R.G.
Neurosci Lett. 1990 Mar 26;111(1-2):58-63. doi: 10.1016/0304-3940(90)90344-9.
Retinae of early postnatal rabbits were enzymatically dissociated and explanted in a culture system. The prospective myelinated region was discarded in order to avoid the presence of astrocytic or mesenchymal cells. After about 14 days in vitro (DIV), outgrowing glial (Müller) cells formed what light optically appeared to be confluent monolayers but by electron microscopy was shown to consist of flat epithelioid cells which overlapped considerably by extension of cytoplasmic tongues. Applying the freeze-fracture technique, apposed membranes of these cells were demonstrated to express infrequently but consistently both gap and tight junctions. This kind of junctions has never been observed on the membrane of rabbit Müller cells in situ. In comparison with Müller cell membranes in situ, the density of intramembrane particles was considerably reduced. Orthogonal arrays of particles which are characteristic elements of Müller cells in situ were not detected. Our results suggest that in homogeneous cell culture, Müller cells form some kind of epithelium-like specialized intercellular junctions. This situation resembles that of closely related glial cell types which form homogeneous layers in situ as e.g. retinal pigment epithelium cells expressing tight junctions, and marginal astrocytes being coupled by extensive gap junctions.
将出生后早期兔子的视网膜进行酶解,并接种于培养体系中。为避免星形细胞或间充质细胞的存在,将预期的髓鞘形成区域去除。体外培养约14天后(体外培养天数),生长出的神经胶质(穆勒)细胞形成了光学显微镜下看似融合的单层细胞,但电子显微镜显示其由扁平上皮样细胞组成,这些细胞通过细胞质舌状突起大量重叠。应用冷冻蚀刻技术,证明这些细胞的相邻膜偶尔但持续表达缝隙连接和紧密连接。这种连接在原位兔穆勒细胞膜上从未观察到。与原位穆勒细胞膜相比,膜内颗粒的密度显著降低。未检测到原位穆勒细胞特有的颗粒正交排列。我们的结果表明,在同质细胞培养中,穆勒细胞形成了某种上皮样的特殊细胞间连接。这种情况类似于密切相关的神经胶质细胞类型,它们在原位形成同质层,例如表达紧密连接的视网膜色素上皮细胞,以及通过广泛的缝隙连接相连的边缘星形细胞。