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非洲爪蟾中与神经元细胞周期蛋白依赖性激酶(cdk5)同源的蛋白在原肠胚形成期的胚胎中表达。

The Xenopus laevis homologue to the neuronal cyclin-dependent kinase (cdk5) is expressed in embryos by gastrulation.

作者信息

Gervasi C, Szaro B G

机构信息

Department of Biological Sciences, State University of New York, Albany 12222, USA.

出版信息

Brain Res Mol Brain Res. 1995 Nov;33(2):192-200. doi: 10.1016/0169-328x(95)00109-6.

Abstract

Phosphorylation of the neuronal cytoskeletal proteins NF-H, NF-M and tau is important for normal axonal development, and is involved in axonal injury and neurodegenerative diseases. In mammalian neurons, one kinase that phosphorylates these axonal cytoskeletal proteins is cyclin-dependent kinase 5 (cdk5). Cdk5 is a member of the family of cyclin-dependent kinases (cdks), whose other family members regulate mitosis. Unlike the other cdks, cdk5 is abundant in differentiated neurons. Embryos of the clawed frog Xenopus laevis have proved useful for studying other cyclin-dependent kinases, neurofilament proteins and tau during development. As a first step in studying the role of cdk5 and its effects on neurofilaments during Xenopus neural development, four cDNA clones were isolated by screening a frog brain cDNA library at lowered stringency with a cDNA probe to rat cdk5. The frog cdk5 clones encoded a protein of 292 amino acids that was 97% identical to rat cdk5. In situ hybridization demonstrated that the Xenopus cdk5 transcript, like that of mammals, was expressed in differentiated post-mitotic neurons. The high degree of sequence homology and shared neuronal expression suggests that the role of cdk5 in neurons is highly conserved between mammals and amphibians. Northern blot analysis indicated that during Xenopus development, cdk5 mRNA was first expressed between the midblastula transition and gastrulation, which both occur long before neuronal differentiation. These stages mark the transition from synchronous to asynchronous cell division and are the earliest stages of zygotic gene expression. This early expression of Xenopus cdk5 mRNA implies a role for cdk5 during embryogenesis that is separate from its role as an axonal cytoskeletal protein kinase. These observations provide the foundation for exploiting X. laevis embryos to study the role of cdk5 both in the early stages of axonal differentiation and also in early embryogenesis.

摘要

神经元细胞骨架蛋白NF-H、NF-M和tau的磷酸化对于正常轴突发育很重要,并且与轴突损伤和神经退行性疾病有关。在哺乳动物神经元中,一种使这些轴突细胞骨架蛋白磷酸化的激酶是细胞周期蛋白依赖性激酶5(cdk5)。Cdk5是细胞周期蛋白依赖性激酶(cdks)家族的成员,该家族的其他成员调节有丝分裂。与其他cdks不同,cdk5在分化的神经元中含量丰富。爪蟾非洲爪蟾的胚胎已被证明有助于研究发育过程中的其他细胞周期蛋白依赖性激酶、神经丝蛋白和tau。作为研究cdk5在非洲爪蟾神经发育过程中的作用及其对神经丝影响的第一步,通过用大鼠cdk5的cDNA探针在较低严谨度下筛选蛙脑cDNA文库,分离出了四个cDNA克隆。蛙cdk5克隆编码一种292个氨基酸的蛋白质,与大鼠cdk5的同源性为97%。原位杂交表明,非洲爪蟾cdk5转录本与哺乳动物的一样,在有丝分裂后分化的神经元中表达。高度的序列同源性和共同的神经元表达表明,cdk5在神经元中的作用在哺乳动物和两栖动物之间高度保守。Northern印迹分析表明,在非洲爪蟾发育过程中,cdk5 mRNA最早在囊胚中期转变和原肠胚形成之间表达,这两个阶段都发生在神经元分化之前很久。这些阶段标志着从同步细胞分裂到异步细胞分裂的转变,是合子基因表达的最早阶段。非洲爪蟾cdk5 mRNA的这种早期表达意味着cdk5在胚胎发生过程中具有与其作为轴突细胞骨架蛋白激酶的作用不同的作用。这些观察结果为利用非洲爪蟾胚胎研究cdk5在轴突分化早期阶段以及早期胚胎发生中的作用奠定了基础。

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