Frotscher M, Zimmer J
J Comp Neurol. 1987 May 8;259(2):266-76. doi: 10.1002/cne.902590207.
Glutamate decarboxylase (GAD) immunocytochemistry was used to study GABAergic neurons and synapses in intracerebral allografts of the rat hippocampus and fascia dentata. Tissue blocks of regio inferior of Ammon's horn (hippocampal field CA3) or of the fascia dentata were taken from newborn rats and transplanted to the hippocampal region of young adult rats. After 6 1/2 months' survival the recipient brains were fixed by perfusion and serially sectioned on a Vibratome. Sections containing the transplant and/or the host hippocampal region were immunostained for GAD and flat-embedded in Araldite for a correlated light and electron microscopic analysis. Immunostained neurons and terminals in the transplants were compared to immunoreactive elements in the hippocampus and fascia dentata of the hosts and other, normal rats. As in the hippocampal formation in situ, GAD-immunoreactive neurons and terminals in the transplants were observed in all layers. In dentate transplants a preponderance of immunostained cells was found just beneath the granule cell layer. In both hippocampal and dentate transplants, immunoreactive terminals were most abundant in the cell layers where they formed characteristic pericellular baskets around the pyramidal and granule cell bodies. In the electron microscope, the transplant GAD-immunoreactive neurons exhibited numerous cytoplasmic organelles, deeply infolded nuclei, and nuclear rods. Immunoreactive terminals formed symmetric synaptic contacts on the cell bodies, dendritic shafts, and spines of transplant pyramidal cells, granule cells, and hilar neurons. These are normal characteristics of GAD-immunoreactive neurons and terminals as also observed in the hippocampus of the host rats and the normal controls. Our results demonstrate that GABAergic neurons survive transplantation and develop a cell-specific morphology that includes the axonal projections.
采用谷氨酸脱羧酶(GAD)免疫细胞化学方法,研究大鼠海马和齿状回脑内同种异体移植组织中的γ-氨基丁酸(GABA)能神经元和突触。取自新生大鼠的海马角下区(海马CA3区)或齿状回组织块,移植到成年幼鼠的海马区。存活6个半月后,通过灌注固定受体脑,并在振动切片机上连续切片。对含有移植组织和/或宿主海马区的切片进行GAD免疫染色,并平嵌入环氧树脂中,进行相关的光镜和电镜分析。将移植组织中免疫染色的神经元和终末与宿主以及其他正常大鼠海马和齿状回中的免疫反应性成分进行比较。与原位海马结构一样,在移植组织的所有层中均观察到GAD免疫反应性神经元和终末。在齿状回移植组织中,在颗粒细胞层下方发现大量免疫染色细胞。在海马和齿状回移植组织中,免疫反应性终末在细胞层中最为丰富,它们在锥体细胞和颗粒细胞体周围形成特征性的细胞周篮状结构。在电子显微镜下,移植组织中GAD免疫反应性神经元表现出众多的细胞质细胞器、深陷的细胞核和核棒。免疫反应性终末在移植的锥体细胞、颗粒细胞和门区神经元的细胞体、树突干和棘上形成对称的突触联系。这些都是GAD免疫反应性神经元和终末的正常特征,在宿主大鼠海马和正常对照中也观察到。我们的结果表明,GABA能神经元在移植后存活,并发育出包括轴突投射在内的细胞特异性形态。