Marchisio P C, Cirillo D, Naldini L, Calissano P
J Neurocytol. 1981 Feb;10(1):45-55. doi: 10.1007/BF01181744.
The distribution of nerve growth factor (NGF) in chick embryo sympathetic neurons has been followed by two distinct procedures, that is, indirect immunofluorescence microscopy employing purified NGF antibodies and autoradiography after exposure of cells to [125I]NGF. This study shows that NGF bound to sympathetic neurons is not uniformly distributed but appears in spots over the surface of perikarya and along processes. If the same cells, after incubation with NGF and fixation, are treated with methanol-acetone to allow permeation of immunoglobulins across the plasma membrane, NGF antibody immunoreactive material is also found within the cytoplasm and notably in the paranuclear area only in sympathetic neurons. Analogous findings are obtained when sympathetic neurons are incubated in the presence of [125I]NGF, fixed, sectioned and processed for autoradiography. Also with this technique NGF appears to be localized in the cytoplasmic compartment and is found around the nucleus. These studies are discussed in connection with the results of similar experiments performed on a clonal line of NGF target cells known as PC12.
通过两种不同的方法追踪了鸡胚交感神经元中神经生长因子(NGF)的分布,即使用纯化的NGF抗体进行间接免疫荧光显微镜检查,以及在细胞暴露于[125I]NGF后进行放射自显影。这项研究表明,与交感神经元结合的NGF并非均匀分布,而是出现在核周体表面和沿着突起的斑点中。如果将与NGF孵育并固定后的相同细胞用甲醇 - 丙酮处理,以使免疫球蛋白透过质膜,那么在细胞质内,特别是仅在交感神经元的核旁区域也能发现NGF抗体免疫反应性物质。当交感神经元在[125I]NGF存在下孵育、固定、切片并进行放射自显影处理时,也能得到类似的结果。使用这种技术,NGF似乎也定位在细胞质区室中,并在细胞核周围被发现。结合对一种称为PC12的NGF靶细胞克隆系进行的类似实验结果,对这些研究进行了讨论。