Sarthy P V, Bacon W
Dev Biol. 1985 Dec;112(2):284-91. doi: 10.1016/0012-1606(85)90399-9.
In order to examine the appearance of synaptic vesicles and to correlate it with the formation of the synaptic layers, we have determined the staining pattern of a murine monoclonal antibody (SV 48) to a synaptic vesicle-associated protein in developing rat retina. The antigen was detected by the indirect immunofluorescence technique using cryostat sections of paraformaldehyde-fixed retinas. In the adult retina, the antibody stained both the outer plexiform (OPL) and the inner plexiform layers (IPL). The nuclear layers and the nerve fiber layer (NFL) were devoid of any staining. In prenatal and early postnatal (P) retinas, the antibody stained two bands which corresponded to the respective locations of the NFL and IPL. Staining in the NFL increased until P-4 and began to decline subsequently, and by P-8 little staining was left in this layer. In contrast, in the IPL, the intensity of staining increased gradually and leveled off by P-10. In the outer retina, a band of fluorescence corresponding to the OPL was first observed at P-5 and increased in intensity up to P-10. Immunoblotting studies showed that the major immunoreactive material from adult and embryonic retinas had a Mr approximately 65,000-67,000. As expected from its developmental pattern, all bands appeared initially in the central retina and subsequently in the peripheral retina. Our results show that the synaptic vesicle-protein is present in the nerve fiber layer before synaptogenesis in the central nervous system. Subsequently, the protein is lost from the NFL, possibly as a consequence of synapse formation.
为了研究突触小泡的外观并将其与突触层的形成相关联,我们确定了一种针对发育中大鼠视网膜突触小泡相关蛋白的鼠单克隆抗体(SV 48)的染色模式。使用多聚甲醛固定视网膜的低温切片,通过间接免疫荧光技术检测抗原。在成年视网膜中,该抗体对外部神经丛层(OPL)和内部神经丛层(IPL)均有染色。核层和神经纤维层(NFL)没有任何染色。在产前和产后早期(P)视网膜中,该抗体染出两条带,分别对应于NFL和IPL的各自位置。NFL中的染色在P - 4之前增加,随后开始下降,到P - 8时该层几乎没有染色。相反,在IPL中,染色强度逐渐增加,到P - 10时趋于平稳。在视网膜外层,对应于OPL的一条荧光带在P - 5时首次观察到,强度在P - 10之前增加。免疫印迹研究表明,来自成年和胚胎视网膜的主要免疫反应性物质的Mr约为65,000 - 67,000。正如从其发育模式所预期的那样,所有条带最初出现在中央视网膜,随后出现在周边视网膜。我们的结果表明,突触小泡蛋白在中枢神经系统突触形成之前存在于神经纤维层中。随后,该蛋白从NFL中消失,可能是突触形成的结果。