Calcium signaling mediates proliferation of the precursor cells that give rise to the ciliated left-right organizer in the zebrafish embryo.
Calcium signaling mediates proliferation of the precursor cells that give rise to the ciliated left-right organizer in the zebrafish embryo.
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DOI：
10.3389/fmolb.2023.1292076
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发表时间：
2023
影响因子：
5
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文献类型：
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我们的几个内部器官，包括心脏、肺、胃和脾，沿着左右（LR）身体轴沿着发育。在早期胚胎发育过程中建立LR不对称性或内脏偏侧性的错误可能导致出生缺陷。在一些脊椎动物中，包括人类、老鼠、青蛙和鱼类，纤毛在建立器官的偏侧性中起着核心作用。在称为“左右组织者”（LRO）的短暂胚胎结构中的运动纤毛产生定向流体流动，该定向流体流动已被提议由机械感觉纤毛检测以触发定向LR轴的不对称信号传导途径。然而，控制纤毛LRO的形式和功能的机制仍然知之甚少。在斑马鱼胚胎中，称为背侧前体细胞（DFC）的前体细胞发育成称为Kupffer囊泡（KV）的瞬时纤毛结构，其功能为LRO。DFC可以在胚胎中可视化和跟踪，从而为研究控制LRO发育的机制提供了机会。以前的工作表明，通过有丝分裂的DFC的增殖是开发功能性KV的关键步骤。在这里，我们进行了有针对性的药理学筛选，以确定控制DFC增殖的机制。肌质/内质网Ca ~（2+）-ATP酶（SERCA）的小分子抑制剂被发现减少DFC有丝分裂。SERCA泵参与调节细胞内钙离子（Ca 2+）浓度。为了使活胚胎中的Ca 2+可视化，我们使用荧光Ca 2+生物传感器GCaMP 6 f产生转基因斑马鱼。实时成像确定了明确发生在DFC中的动态细胞质Ca 2+瞬变（“通量”）。此外，我们报告的Ca 2+通量事件发生在DFCs的核。细胞核Ca ~（2+）流发生在即将进行有丝分裂的DFC中。我们发现，在DFC增殖阶段的SERCA抑制剂治疗改变Ca 2+动力学，减少KV纤毛细胞的数量，并改变胚胎偏侧性。从机制上讲，SERCA抑制剂处理消除了细胞质和细胞核的Ca 2+通量事件，并减少了DFC通过细胞周期S/G2期的进展。这些结果确定了SERCA介导的Ca 2+信号作为产生纤毛LRO的前体细胞的有丝分裂调节因子。
Several of our internal organs, including heart, lungs, stomach, and spleen, develop asymmetrically along the left-right (LR) body axis. Errors in establishing LR asymmetry, or laterality, of internal organs during early embryonic development can result in birth defects. In several vertebrates—including humans, mice, frogs, and fish—cilia play a central role in establishing organ laterality. Motile cilia in a transient embryonic structure called the “left-right organizer” (LRO) generate a directional fluid flow that has been proposed to be detected by mechanosensory cilia to trigger asymmetric signaling pathways that orient the LR axis. However, the mechanisms that control the form and function of the ciliated LRO remain poorly understood. In the zebrafish embryo, precursor cells called dorsal forerunner cells (DFCs) develop into a transient ciliated structure called Kupffer’s vesicle (KV) that functions as the LRO. DFCs can be visualized and tracked in the embryo, thereby providing an opportunity to investigate mechanisms that control LRO development. Previous work revealed that proliferation of DFCs via mitosis is a critical step for developing a functional KV. Here, we conducted a targeted pharmacological screen to identify mechanisms that control DFC proliferation. Small molecule inhibitors of the sarcoplasmic/endoplasmic reticulum Ca2+-ATPase (SERCA) were found to reduce DFC mitosis. The SERCA pump is involved in regulating intracellular calcium ion (Ca2+) concentration. To visualize Ca2+ in living embryos, we generated transgenic zebrafish using the fluorescent Ca2+ biosensor GCaMP6f. Live imaging identified dynamic cytoplasmic Ca2+ transients (“flux”) that occur unambiguously in DFCs. In addition, we report Ca2+ flux events that occur in the nucleus of DFCs. Nuclear Ca2+ flux occurred in DFCs that were about to undergo mitosis. We find that SERCA inhibitor treatments during DFC proliferation stages alters Ca2+ dynamics, reduces the number of ciliated cells in KV, and alters embryo laterality. Mechanistically, SERCA inhibitor treatments eliminated both cytoplasmic and nuclear Ca2+ flux events, and reduced progression of DFCs through the S/G2 phases of the cell cycle. These results identify SERCA-mediated Ca2+ signaling as a mitotic regulator of the precursor cells that give rise to the ciliated LRO.
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影响因子：
64.8
作者：
Essner, JJ;Vogan, KJ;Brueckner, M
通讯作者：
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影响因子：
9.2
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作者：
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