Moody S A, Miller V, Spanos A, Frankfurter A
Department of Anatomy and Neuroscience Program, George Washington University Medical Center, Washington, D.C. 20037, USA.
J Comp Neurol. 1996 Jan 8;364(2):219-30. doi: 10.1002/(SICI)1096-9861(19960108)364:2<219::AID-CNE3>3.0.CO;2-8.
In mammals, there are seven classes of beta-tubulin genes, one of which, class III, is neuron specific. Using class-specific monoclonal antibodies, class III beta-tubulin protein could not be detected in frog embryos or in adults with either Western blotting or immunohistochemical techniques. In contrast, the class II beta-tubulin protein, which is predominant in mammalian brain but is also expressed in other tissues, is expressed only in neurons in frog embryos. Protein was detected only in neurons from late stages of neural tube closure through premetamorphic stages. At stages 21-28, the pioneering axons of Rohon-Beard, commissural, primary motor, and trigeminal ganglion neurons were distinctly stained in the axon scaffolds that they formed in the embryonic brain and the peripheral mesenchyme. Nonneuronal cells, both outside the nervous system and within it (e.g., radial glia, Müller glia, roof plate, and floor plate cells) were not immunoreactive. Throughout swimming and premetamorphic stages, neuronal cells in all brain regions became immunoreactive as they differentiated and extended axons. Whereas many embryonic neurons became postmitotic during gastrulation stages, neurons expressed detectable levels of class II beta-tubulin protein only beginning at the onset of overt axon outgrowth. These observations demonstrate that the neuron-specific beta-tubulin in frog is a different gene from that in mammals, and its protein product is detectable at the time of axonogenesis rather than neurogenesis.
在哺乳动物中,存在七类β-微管蛋白基因,其中III类是神经元特异性的。使用类特异性单克隆抗体,无论是通过蛋白质印迹法还是免疫组织化学技术,在青蛙胚胎或成体中均检测不到III类β-微管蛋白。相比之下,II类β-微管蛋白在哺乳动物脑中占主导地位,但也在其他组织中表达,在青蛙胚胎中仅在神经元中表达。仅在神经管闭合后期至变态前阶段的神经元中检测到该蛋白。在第21 - 28阶段,罗霍恩 - 比尔神经元、连合神经元、初级运动神经元和三叉神经节神经元的先驱轴突在它们在胚胎脑和外周间充质中形成的轴突支架中被明显染色。神经系统外部和内部的非神经元细胞(例如,放射状胶质细胞、米勒胶质细胞、顶板和底板细胞)没有免疫反应性。在整个游泳和变态前阶段,所有脑区的神经元细胞在分化并延伸轴突时都变得具有免疫反应性。虽然许多胚胎神经元在原肠胚形成阶段进入有丝分裂后期,但神经元仅在明显的轴突生长开始时才表达可检测水平的II类β-微管蛋白。这些观察结果表明,青蛙中神经元特异性的β-微管蛋白与哺乳动物中的是不同的基因,其蛋白质产物在轴突发生时而不是神经发生时可被检测到。