Plaschke M, Nitsch R, Wenzel J, Frotscher M
Institute of Anatomy, University of Freiburg, Germany.
J Comp Neurol. 1992 May 15;319(3):319-36. doi: 10.1002/cne.903190302.
Previous immunocytochemical studies have shown that GABAergic nonpyramidal neurons of the rat hippocampus survive in intracerebral transplants. However, information is still lacking about the dendritic organization and the input synapses of these cells as well as their capacity to express the calcium-binding protein parvalbumin (PARV) under transplant conditions. In the present study, a monoclonal antibody against PARV was used to examine the dendritic morphology and the synaptic organization of parvalbumin-containing GABAergic neurons in hippocampal and dentate transplants. In addition, parvalbumin-containing nonpyramidal neurons were studied in neocortical transplants to compare the differentiation of grafted allocortical and neocortical nonpyramidal neurons. Tissue blocks of hippocampus and fascia dentata and of the parietal neocortex were taken from late embryonic rats (E 21 and E 16, respectively) and were transplanted into a cavity in the somatosensory cortex of young adult rats. After 3.5 or 7 months survival, the recipient brains were fixed by perfusion and immunostained for PARV. As in the hippocampal formation in situ, PARV-containing neurons in the hippocampal transplants were observed within and in the vicinity of the pyramidal and granule cell layer. In neocortical transplants, PARV-immunoreactive cells were distributed in all parts of the transplant with dendrites extending in various directions. In both hippocampal and neocortical transplants, immunoreactive dendrites were smooth and displayed the characteristic regular varicosities known from in situ studies of these cells. Numerous unlabeled terminals as well as a few immunoreactive boutons established synapses on the immunoreactive dendrites. PARV-positive terminals formed the typical pericellular baskets around the immunonegative cell bodies of pyramidal neurons and granule cells in the transplants. They established symmetric synapses with cell bodies and proximal dendrites. Synapses on axon initial segments were absent or rare. Our results demonstrate that allocortical as well as neocortical nonpyramidal neurons transplanted to the neocortex of adult recipients survive transplantation, express the calcium-binding protein parvalbumin, and develop a cell-specific morphology.
以往的免疫细胞化学研究表明,大鼠海马的γ-氨基丁酸能非锥体神经元能在脑内移植中存活。然而,关于这些细胞的树突组织、输入突触以及它们在移植条件下表达钙结合蛋白小白蛋白(PARV)的能力,仍然缺乏相关信息。在本研究中,使用一种抗PARV的单克隆抗体来检查海马和齿状回移植中含小白蛋白的γ-氨基丁酸能神经元的树突形态和突触组织。此外,还对新皮质移植中含小白蛋白的非锥体神经元进行了研究,以比较移植的异皮质和新皮质非锥体神经元的分化情况。海马、齿状回以及顶叶新皮质的组织块取自胚胎晚期大鼠(分别为E21和E16),并移植到成年幼鼠体感皮质的一个腔中。在存活3.5或7个月后,通过灌注固定受体脑,并对PARV进行免疫染色。与原位海马结构一样,在海马移植中,含PARV的神经元在锥体细胞层和颗粒细胞层内及附近被观察到。在新皮质移植中,PARV免疫反应性细胞分布在移植的各个部位,树突向各个方向延伸。在海马和新皮质移植中,免疫反应性树突都很光滑,并显示出原位研究中这些细胞所具有的特征性规则膨体。许多未标记的终末以及一些免疫反应性终扣在免疫反应性树突上形成突触。PARV阳性终末在移植的锥体神经元和颗粒细胞的免疫阴性细胞体周围形成典型的细胞周篮。它们与细胞体和近端树突建立对称突触。轴突起始段上的突触不存在或很少。我们的结果表明,移植到成年受体新皮质的异皮质和新皮质非锥体神经元能够在移植后存活,表达钙结合蛋白小白蛋白,并发育出细胞特异性形态。