Naves F J, Huerta J J, Garcia-Suarez O, Urdangaray N, Esteban I, Del Valle M E, Vega J A
Departamento de Morfología y Biología Celular, Universidad de Oviedo, Spain.
Anat Rec. 1996 Feb;244(2):246-56. doi: 10.1002/(SICI)1097-0185(199602)244:2<246::AID-AR12>3.0.CO;2-P.
The cytoskeleton of mature neurons consists of three main types of filamentous structures: microtubules (or neurotubules) neurofilaments and microfilaments, and of the so-called associated proteins. Neurotubules are formed by alpha- and beta-tubulin; neurofilaments are comprised of three protein subunits (68, 160, and 200 kDa of molecular weight), referred to here as neurofilament proteins (NFPs). The microtubule-associated proteins (MAPs) and tau-proteins form cross bridges between microtubules and other cytoskeletal constituents, as well as cellular organelles. This study analyzes the distribution of several cytoskeletal proteins in adult human dorsal root ganglia (DRG).
Sections of formaldehyde-fixed, paraffin-embedded adult human DRG were processed for PAP immunohistochemistry. Mouse monoclonal antibodies against specific epitopes of alpha- and beta-tubulin, MAP-1, MAP-2, MAP-5, tau-protein, and NFPs (68, 160, and 200 kDa) were used. Furthermore, a quantitative image analysis (optic microdensitometry) was performed to establish the relationship between neuronal size and intensity of immunostaining.
Most of DRG neuron cell bodies displayed immunoreactivity for all assessed antibodies, with the exception of MAP2, which was absent. Nevertheless, the neuronal perikarya showed an heterogeneous pattern of immunoreactivity, which was not related to neuronal profile size. Positive immunolabelling was also observed in satellite cells and Schwann cells for microtubule and MAP1 proteins, and for tau-protein in Schwann cells.
Adult human primary sensory neurons in DRG express immunoreactivity for neurotubule and neurofilament proteins, as well as for some microtubule-associated proteins. However, since large heterogeneity was observed in the expression of those proteins, we conclude that the expression of cytoskeletal proteins is not a criterion to establish DRG neuronal subtypes.
成熟神经元的细胞骨架由三种主要类型的丝状结构组成:微管(或神经微管)、神经丝和微丝,以及所谓的相关蛋白。神经微管由α-和β-微管蛋白形成;神经丝由三个蛋白质亚基(分子量分别为68、160和200 kDa)组成,在此称为神经丝蛋白(NFPs)。微管相关蛋白(MAPs)和tau蛋白在微管与其他细胞骨架成分以及细胞器之间形成交叉桥。本研究分析了几种细胞骨架蛋白在成人背根神经节(DRG)中的分布。
对甲醛固定、石蜡包埋的成人DRG切片进行PAP免疫组织化学处理。使用针对α-和β-微管蛋白、MAP-1、MAP-2、MAP-5、tau蛋白和NFPs(分子量分别为68、160和200 kDa)特定表位的小鼠单克隆抗体。此外,进行了定量图像分析(光学显微密度测定)以确定神经元大小与免疫染色强度之间的关系。
除了不存在的MAP2外,大多数DRG神经元细胞体对所有评估抗体均显示免疫反应性。然而,神经元胞体显示出异质性的免疫反应模式,这与神经元轮廓大小无关。在卫星细胞和施万细胞中也观察到微管和MAP1蛋白以及施万细胞中tau蛋白的阳性免疫标记。
DRG中的成人初级感觉神经元对神经微管和神经丝蛋白以及一些微管相关蛋白表达免疫反应性。然而,由于在这些蛋白的表达中观察到很大的异质性,我们得出结论,细胞骨架蛋白的表达不是建立DRG神经元亚型的标准。