Yoshida Reiko, Kusakabe Takehiro, Kamatani Megumi, Daitoh Motonari, Tsuda Motoyuki
Department of Life Science, Graduate School of Science, Himeji Institute of Technology, Kamigori, Hyogo, Japan.
Zoolog Sci. 2002 Oct;19(10):1079-88. doi: 10.2108/zsj.19.1079.
Heterotrimeric G proteins play crucial roles as mediators of signaling by many extracellular stimuli. The receptors that activate G proteins constitute the largest and most diverse family of cell surface molecules involved in signal transmission of metazoan cells. To investigate G protein signaling in the central nervous system (CNS) of chordates, we isolated cDNA fragments encoding five different G protein alpha subunits (CiGalpha(x), CiGalpha(q), CiGalpha(i1a), CiGalpha(i1b), and CiGalpha(i2)) from larvae of the ascidian, a simple chordate, Ciona intestinalis. In situ hybridization analysis revealed that each isoform had distinct patterns of spatial distribution in embryos. Among them, CiGalpha(i1a) and CiG alpha(i1b) mRNAs were specifically expressed in the CNS of the larva, whereas CiGalpha(q) transcripts were expressed in small parts of the trunk epidermis and the tip of the tail, but not in the CNS. The CiGalpha(x) expression was widely observed throughout the trunk and tail of the embryos, and the signals were stronger in the epidermis, mesenchyme, and tail muscle cells. Comparison of cDNA sequences and the exon-intron organization indicate that CiGalpha(i1a) and CiGalpha(i1b) are produced by alternative splicing of transcripts from a single gene, CiGalpha(i1). In the cleavage and gastrula stages, transcripts of CiGalpha(i1) were widely distributed in embryos, and the expression then became restricted to the CNS of tailbud embryos and larvae. An exhaustive search has failed to find transducin-type alpha subunits in C. intestinalis. Since CiGalpha(i1) is expressed in the ocellus, CiGalpha(i1) may mediate signals from Ci-opsin1, a visual pigment of the ocellus photoreceptor cells.
异源三聚体G蛋白作为许多细胞外刺激信号传导的介质发挥着关键作用。激活G蛋白的受体构成了后生动物细胞信号传递中最大且最多样化的细胞表面分子家族。为了研究脊索动物中枢神经系统(CNS)中的G蛋白信号传导,我们从简单的脊索动物海鞘(Ciona intestinalis)的幼虫中分离出编码五种不同G蛋白α亚基(CiGα(x)、CiGα(q)、CiGα(i1a)、CiGα(i1b)和CiGα(i2))的cDNA片段。原位杂交分析表明,每种异构体在胚胎中具有不同的空间分布模式。其中,CiGα(i1a)和CiGα(i1b) mRNA在幼虫的中枢神经系统中特异性表达,而CiGα(q)转录本在躯干表皮的小部分和尾尖表达,但不在中枢神经系统中表达。CiGα(x)的表达在胚胎的整个躯干和尾部广泛观察到,并且在表皮、间充质和尾肌细胞中的信号更强。cDNA序列和外显子-内含子组织的比较表明,CiGα(i1a)和CiGα(i1b)是由来自单个基因CiGα(i1)的转录本的可变剪接产生的。在卵裂期和原肠胚期,CiGα(i1)的转录本广泛分布于胚胎中,然后表达局限于尾芽胚胎和幼虫的中枢神经系统。详尽的搜索未能在肠海鞘中找到转导素型α亚基。由于CiGα(i1)在眼点中表达,CiGα(i1)可能介导来自Ci-视蛋白1(眼点光感受器细胞的一种视觉色素)的信号。