Expression and localization of the synapse specific proteins in central nervous system of the postnatal rat.

1.突触后密度特异性蛋白（Postsynaptic density specific protein，82 K）是从大鼠脑突触体中提取的突触连接复合物（synaptic junctional complex，SJC）的一种组分，以其为抗原制备单克隆抗体，用于检测SJC的组成蛋白。获得抗体SJ-8，其在免疫印迹分析中识别SJC制剂中分子量为82，000道尔顿的蛋白质（82 K蛋白质）。用免疫组织化学方法研究了82 K蛋白在大鼠小脑中的定位。SJ-8抗体标记浦肯野细胞和颗粒细胞的外周区。在分子层中也用SJ-8染色了小的点状区域。用大鼠脑突触体检测蛋白质的细胞内定位。免疫电镜观察发现SJ-8可强烈标记突触后致密物，并可标记突触后致密物上的纤维网，但不标记突触前膜和突触后膜.突触素表达 ...更多信息 在大鼠小脑中研究了突触素在围产期大鼠小脑皮质中的表达和细胞内定位。出生后1-7天的大鼠通过经心脏灌注4%多聚甲醛固定。将小脑皮质解剖出来，用振动切片机切成40 μ m厚的切片。用1%牛血清白蛋白封闭后，将切片与抗突触素的抗体一起孵育。然后，使切片与FITC缀合的抗小鼠IgG反应，并用配备有落射荧光的光学显微镜检查。围产期大鼠的标本未显示任何阳性反应。生后3天，浦肯野细胞和颗粒细胞周围出现数个阳性斑点。在分子层中也观察到反应的小斑点。这些阳性反应在出生后7天显著增加。2周后小脑皮质的反应模式与成年大鼠相似。阳性反应的发生可能与小脑皮质的突触发生有关。突触体蛋白可能在突触发生前通过轴突从核周体转运到神经末梢。免疫细胞化学检查蛋白质的转运系统。少

1. Postsynaptic density specific protein, 82K proteinA fraction of synaptic junctional complex (SJC) was prepared from rat brain synaptosomes and served as an antigen material to produce monoclonal antibodies for examining the component proteins of the SJC. An antibody, SJ-8, was obtained, which recognized a protein with a molecular weight of 82,000 dalton (82K protein) in the SJC preparation in immunoblotting analysis. The immunohistochemical localization of the 82K protein was studied with the rat cerebellum. The antibody SJ-8 labeled the peripheral area of the Purkinje and granule cells. Small punctate areas were also stained in the molecular layer with SJ-8. Intracellular localization of the protein was examined with the rat brain synaptosomes. Immunoelectron microscopy demonstrated that SJ-8 strongly labeled the postsynaptic density and also a fibrous network spreading out of it. However, the antibody did not label the pre- and postsynaptic membrane.2. Expression of synaptophysin … More in the rat cerebellumExpression and intracellular localization of synaptophysin was examined in cerebellar cortex of the perinatal rat. Rats, 1-7 days after birth, were fixed by perfusion via the heart with 4% paraformaldehyde. The cerebellar cortex was dissected out and cut into 40 mum thick slices with a vibratome. After blocking with 1% bovine serum albumin, the slices were incubated with the antibody against synaptophysin. Then, slices were reacted with FITC- conjugated anti-mouse IgG and examined with a light microscopy equipped with epifluorescence. Specimens from perinatal rats did not show any positive reactions. Several positive spots appeared around the Purkinje cells and granule cells three days after birth. Small punctates of reaction were also observed in the molecular layer. These positive reactions significantly increased 7 days after birth. The reaction pattern in the cerebellar cortex after 2 weeks was similar to that of adult rats. The development of positive reaction may involve the synaptogenesis in the cerebellar cortex. Synaptophysin might be transported from the perikaryon to the nerve ending via the axon before the synaptogenesis. The transporting system of the protein is immunocytochemically examined. Less