Vanhems E, Delbos M, Geffard M, Viellemaringe J
Laboratoire de Neuroendocrinologie, URA CNRS 1138, Université Bordeaux I, Talence, France.
Neuroscience. 1994 Feb;58(3):649-55. doi: 10.1016/0306-4522(94)90088-4.
Organotypic cultures established from the third thoracic ganglion of locust embryo have been used to investigate dopamine receptors. In this in vitro system, neurites emerge directly from the explants and form a dense network around the explants, presenting cell surface freely exposed for experimental labelling. Polyclonal anti-idiotypic antibodies raised in rabbits to antibodies against dopamine conjugate, and previously found to bind to dopamine receptors, have been used to investigate putative dopamine receptors in these neurites. Immunocytochemical detection by light microscopy employing immunofluorescence labelling, was correlated with electron microscopy, using peroxidase staining. In addition to a location for dopamine receptors on the neurite surface, intracellular binding sites were also found in neurites. This internal labelling might represent an intracellular pool of dopamine receptor precursors. The labelling was specific in that it was not present when the anti-idiotypic dopamine antibodies were replaced with non-immune serum or when preincubation with conjugated dopamine preceded incubation with anti-idiotypic dopamine antibodies.
从蝗虫胚胎第三胸神经节建立的器官型培养物已被用于研究多巴胺受体。在这个体外系统中，神经突直接从外植体中长出，并在外植体周围形成密集网络，使细胞表面可自由暴露用于实验标记。用兔产生的针对多巴胺偶联物抗体的多克隆抗独特型抗体，先前发现其可与多巴胺受体结合，已被用于研究这些神经突中假定的多巴胺受体。通过免疫荧光标记的光学显微镜进行免疫细胞化学检测，并与使用过氧化物酶染色的电子显微镜相关联。除了在神经突表面存在多巴胺受体外，在神经突中还发现了细胞内结合位点。这种内部标记可能代表多巴胺受体前体的细胞内池。这种标记是特异性的，因为当用非免疫血清替代抗独特型多巴胺抗体时，或者在用抗独特型多巴胺抗体孵育之前先用偶联多巴胺预孵育时，标记不存在。
