Marusich M F, Furneaux H M, Henion P D, Weston J A
Institute of Neuroscience, University of Oregon, Eugene 97403.
J Neurobiol. 1994 Feb;25(2):143-55. doi: 10.1002/neu.480250206.
We have utilized immunochemical techniques to investigate the developmental expression of the Hu proteins, a neuron-specific family of RNA binding proteins in vertebrates. Previous work suggests that these proteins may play an important role in neuronal development and maintenance. For the present study, we developed a monoclonal antibody (MAb 16A11) that binds specifically to an epitope present in gene products of all known Hu genes, including HuD, HuC, and Hel-N1. Using brief pulses (1-2 h) of the DNA precursor analog bromodeoxyuridine (BrdU) in conjunction with MAb 16A11, we observed Hu+/BrdU+ cells in nascent sensory and sympathetic ganglia in vivo, and in populations of cultured neural crest cells. In addition, a few Hu+ cells were ambiguously BrdU+ in the neural tube. We conclude that Hu+ cells first appear in avian neurogenic populations immediately before neuronal birthdays in the peripheral nervous system, and at the time of withdrawal from the mitotic cycle in the central nervous system. Consistent with these conclusions, we have also observed neural crest-derived cells that are both Hu+ and in metaphase of the cell cycle. We suggest that Hu proteins function early in neurogenic differentiation.
我们利用免疫化学技术研究了Hu蛋白的发育表达情况,Hu蛋白是脊椎动物中一类神经元特异性的RNA结合蛋白家族。先前的研究表明,这些蛋白可能在神经元的发育和维持过程中发挥重要作用。在本研究中,我们制备了一种单克隆抗体(MAb 16A11),它能特异性结合所有已知Hu基因(包括HuD、HuC和Hel-N1)产物中存在的一个表位。通过将DNA前体类似物溴脱氧尿苷(BrdU)短暂脉冲处理(1 - 2小时)并结合MAb 16A11,我们在新生的感觉和交感神经节体内以及培养的神经嵴细胞群体中观察到了Hu⁺/BrdU⁺细胞。此外,神经管中有少数Hu⁺细胞呈模糊的BrdU⁺状态。我们得出结论,Hu⁺细胞最早出现在鸟类神经源性群体中,在外周神经系统中紧接神经元诞生之前,在中枢神经系统中则是在退出有丝分裂周期之时。与这些结论一致,我们还观察到了既是Hu⁺又是处于细胞周期中期的神经嵴衍生细胞。我们认为Hu蛋白在神经源性分化早期发挥作用。