Barnett M W, Old R W, Jones E A
Department of Biological Sciences, University of Warwick, Coventry, UK.
Dev Growth Differ. 1998 Feb;40(1):47-57. doi: 10.1046/j.1440-169x.1998.t01-5-00006.x.
Two natural neural inducing sources have been used, the notochord and the somites together with the growth factor bFGF, to investigate the anterior/posterior patterning of neural tissue in an animal cap explant model in Xenopus laevis. Notochord and somite tissue from stages 12.5/13 and 16, respectively, were manually isolated, and combined heterochronically with responding animal cap ectoderm aged to gastrula stages. Somite recombinants were also constructed with animal caps injected with noggin mRNA. The responses of the ectoderm were analyzed by reverse transcription polymerase chain reaction (RT-PCR) detection of marker gene expression, and in some cases by in situ hybridization. The requirement for FGF receptor function was analyzed using the dominant negative FGF receptor (XFD). The experiments showed that bFGF is capable of direct neural induction in caps aged to stage 10.5. It was also shown that notochords are capable of inducing anterior neural tissue in gastrula stage animal cap ectoderm, and this induction is sensitive to XFD in the responding tissue. Injection of noggin mRNA results in the induction of anterior neural differentiation, and it was demonstrated that this induction was insensitive to the expression of XFD in the responding tissue. It was also shown that somite tissue recombined with gastrula stage animal cap ectoderm, can induce both anterior and posterior nervous tissue and can also posteriorize noggin-induced anterior neural tissue when combined with noggin-injected animal cap ectoderm. This response is partially sensitive to XFD expression. The results shed light on the role of competence of animal cap ectoderm and the signals from postgastrulation axial and paraxial mesoderm in the patterning of the amphibian nervous system.
在非洲爪蟾的动物帽外植体模型中,已使用两种天然神经诱导源,即脊索和体节以及生长因子碱性成纤维细胞生长因子(bFGF),来研究神经组织的前后模式形成。分别从12.5/13期和16期手动分离出脊索和体节组织,并与已发育到原肠胚期的反应性动物帽外胚层进行异时组合。还构建了体节重组体,即将动物帽注射noggin信使核糖核酸(mRNA)。通过逆转录聚合酶链反应(RT-PCR)检测标记基因表达来分析外胚层的反应,在某些情况下还通过原位杂交进行分析。使用显性负性成纤维细胞生长因子受体(XFD)分析对成纤维细胞生长因子受体功能的需求。实验表明,bFGF能够在发育到10.5期的帽中直接诱导神经。还表明,脊索能够在原肠胚期动物帽外胚层中诱导前神经组织,并且这种诱导在反应组织中对XFD敏感。注射noggin mRNA会导致前神经分化的诱导,并且已证明这种诱导对反应组织中XFD的表达不敏感。还表明,与原肠胚期动物帽外胚层重组的体节组织,既能诱导前神经组织又能诱导后神经组织,并且当与注射noggin的动物帽外胚层组合时,还能使noggin诱导的前神经组织向后分化。这种反应对XFD表达部分敏感。这些结果揭示了动物帽外胚层的感受态作用以及原肠胚后期轴向和旁轴中胚层发出的信号在两栖动物神经系统模式形成中的作用。