Molecular and functional characterization of inositol trisphosphate receptors during early zebrafish development
Molecular and functional characterization of inositol trisphosphate receptors during early zebrafish development
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DOI：
10.1074/jbc.m700940200
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发表时间：
2007-05-11
影响因子：
4.8
通讯作者：
Patel, Sandip
中科院分区：
文献类型：
--
作者：
Ashworth, Rachel;Devogelaere, Benoit;Patel, Sandip
胞质内Ca2+的波动对包括发育的许多方面在内的各种细胞过程至关重要。通过肌醇三磷酸（IP3）的产生和随后IP3敏感的Ca2+通道的激活，细胞内Ca2+储存的动员是一种普遍存在的手段，通过各种刺激介导其细胞效应。虽然IP3受体在受精过程中已经得到了很好的研究，但关于它们在随后发育过程中可能参与的信息却很少。在本研究中，我们研究了IP3受体在斑马鱼早期发育中的作用。我们报道了斑马鱼IP3受体的首次分子分析，表明斑马鱼基因组像哺乳动物一样含有三种不同的IP3受体基因。所有同工异构体的mRNA在64细胞阶段都可以检测到不同水平，并且ip3诱导的Ca2+瞬态可以在体内裂解期间以受控的方式容易产生（通过闪光光解）。此外，我们发现早期囊胚的形成被IP3受体或磷脂酶C的药理学阻断，通过注射IRBIT （IP3受体结合蛋白与IP3释放）对前者的分子抑制，以及在第二个细胞周期完成后耗尽thapsigargin敏感的Ca2+储存而中断。然而，抑制Ca2+进入或ryanodine受体几乎没有影响。我们的工作定义了IP3受体在遗传和光学可塑性模型脊椎动物生物早期发育中的重要性。
Fluctuations in cytosolic Ca2+ are crucial for a variety of cellular processes including many aspects of development. Mobilization of intracellular Ca2+ stores via the production of inositol trisphosphate (IP3) and the consequent activation of IP3-sensitive Ca2+ channels is a ubiquitous means by which diverse stimuli mediate their cellular effects. Although IP3 receptors have been well studied at fertilization, information regarding their possible involvement during subsequent development is scant. In the present study we examined the role of IP3 receptors in early development of the zebrafish. We report the first molecular analysis of zebrafish IP3 receptors which indicates that, like mammals, the zebrafish genome contains three distinct IP3 receptor genes. mRNA for all isoforms was detectable at differing levels by the 64 cell stage, and IP3-induced Ca2+ transients could be readily generated (by flash photolysis) in a controlled fashion throughout the cleavage period in vivo. Furthermore, we show that early blastula formation was disrupted by pharmacological blockade of IP3 receptors or phospholipase C, by molecular inhibition of the former by injection of IRBIT (IP3 receptor-binding protein released with IP3) and by depletion of thapsigargin-sensitive Ca2+ stores after completion of the second cell cycle. Inhibition of Ca2+ entry or ryanodine receptors, however, had little effect. Our work defines the importance of IP3 receptors during early development of a genetically and optically tractable model vertebrate organism.