Expression patterns of Src-family tyrosine kinases during Xenopus laevis development.

Zoltán Ferjentsik, Radek Sindelka, J. Jonák
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引用次数: 4

Abstract

Src family tyrosine kinases (SFKs) play important roles in cell morphology, differentiation, motility and proliferation. Elevated expression and/or specific activity of Src kinases are characteristic for several types of human cancer. However, little information is available about the role and spatio-temporal expression of SFKs in early embryonic development. In this study we characterized, in Xenopus laevis, the expression patterns of five SFK genes src, fyn, yes, lyn and laloo as well as of the csk gene, a negative regulator of SFKs, using RT-qPCR and in situ hybridisation. We found that transcripts of all SFKs and csk were already detectable in one-cell embryos and their levels similarly oscillated during subsequent development. First, after stage 8, the levels of SFK and csk mRNAs began to decrease, reached a minimum between stages 10 and 28 and increased again. In the later stages (33-45), the levels of fyn, yes and csk mRNAs returned to approximately maternal ones, whereas the src, laloo and lyn mRNA transcripts exceeded, up to about 3.5-6-fold, their maternal levels. In situ hybridisation analysis located the SFK and csk transcripts in the animal hemisphere of Xenopus embryos. Subsequent gastrula stages showed signals in ectodermal cells, mid-neurula stage embryos at neural folds, and the tailbud stages showed strong signals in the brain and neural tube. RT-qPCR concentration profiling along the animal-vegetal axis proved in blastula and gastrula the preferential localisation of yes, src, lyn and csk transcripts towards the animal pole in a gradient-like manner. In contrast, laloo and fyn displayed a vegetal pole preference.
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非洲爪蟾发育过程中src家族酪氨酸激酶的表达模式
Src家族酪氨酸激酶(SFKs)在细胞形态、分化、运动和增殖中起重要作用。Src激酶的表达和/或特异性活性升高是几种人类癌症的特征。然而,关于sfk在早期胚胎发育中的作用和时空表达的信息很少。本研究利用RT-qPCR和原位杂交技术,分析了非洲爪蟾SFK的5个基因src、fyn、yes、lyn和laloo以及SFK的负调控基因csk的表达模式。我们发现所有sfk和csk的转录本在单细胞胚胎中已经可以检测到,并且它们的水平在随后的发育过程中类似地振荡。首先,在第8阶段之后,SFK和csk mrna的水平开始下降,在第10和28阶段之间达到最低水平,然后再次上升。在后期(33-45),fyn、yes和csk mRNA水平恢复到母体水平,而src、laloo和lyn mRNA转录本超过母体水平,最高可达3.5-6倍。原位杂交分析在爪蟾胚胎的动物半球中找到了SFK和csk转录本。随后的原胚阶段在外胚层细胞中显示信号,神经鞘中期胚胎在神经褶皱处显示信号,尾芽阶段在脑和神经管中显示强烈信号。沿着动物-植物轴的RT-qPCR浓度分析证明,在囊胚和原胚中,yes、src、lyn和csk转录本以类似梯度的方式优先定位于动物极。相比之下,laloo和fyn表现出植物极偏好。
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