Generation of GCaMP6s-Expressing Zebrafish to Monitor Spatiotemporal Dynamics of Calcium Signaling Elicited by Heat Stress.
Generation of GCaMP6s-Expressing Zebrafish to Monitor Spatiotemporal Dynamics of Calcium Signaling Elicited by Heat Stress.
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生成表达 GCaMP6s 的斑马鱼以监测热应激引起的钙信号传导的时空动态
DOI：
10.3390/ijms22115551
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发表时间：
2021-05-24
影响因子：
5.6
通讯作者：
Cui Z
中科院分区：
文献类型：
--
作者：
Li F;Long Y;Xie J;Ren J;Zhou T;Song G;Li Q;Cui Z
生物体快速感知和传递环境压力信号的能力对它们的生存至关重要。Ca~(2+)是一种多功能的细胞内信使，参与感受各种压力并调节随后的细胞反应。到目前为止，我们对钙信号的了解主要来自体外组织和培养的细胞系，而脊椎动物体内应激触发的钙信号的时空动力学仍有待研究。在这里，我们描述了一个普遍表达GCaMP6s的转基因斑马鱼品系的产生和特征，GCaMP6s是一种遗传编码的钙指示剂(Geci)。我们开发了一种方法来研究热应激诱导的钙事件的时空模式。在发育中的斑马鱼中，暴露在热应激下会立即和瞬间产生钙信号。头部和躯干被层中广泛分布的细胞是第一批反应细胞，不同的细胞群对热应激表现出不同的反应模式。热应激诱导的钙信号在30 S时达到峰值，暴露后迅速下降至接近基础水平。LaCl3和Capsazepine抑制热诱导的钙信号转导，CaMKII(钙调素依赖的蛋白激酶II)和HSF1(热休克因子1)抑制剂处理均显著抑制增强的热休克反应(HSR)。总之，我们描述了热诱导钙信号的时空动态，并证实了钙-CaMKII-HSF1通路在调节斑马鱼HSR中的功能。
The ability of organisms to quickly sense and transduce signals of environmental stresses is critical for their survival. Ca2+ is a versatile intracellular messenger involved in sensing a wide variety of stresses and regulating the subsequent cellular responses. So far, our understanding for calcium signaling was mostly obtained from ex vivo tissues and cultured cell lines, and the in vivo spatiotemporal dynamics of stress-triggered calcium signaling in a vertebrate remains to be characterized. Here, we describe the generation and characterization of a transgenic zebrafish line with ubiquitous expression of GCaMP6s, a genetically encoded calcium indicator (GECI). We developed a method to investigate the spatiotemporal patterns of Ca2+ events induced by heat stress. Exposure to heat stress elicited immediate and transient calcium signaling in developing zebrafish. Cells extensively distributed in the integument of the head and body trunk were the first batch of responders and different cell populations demonstrated distinct response patterns upon heat stress. Activity of the heat stress-induced calcium signaling peaked at 30 s and swiftly decreased to near the basal level at 120 s after the beginning of exposure. Inhibition of the heat-induced calcium signaling by LaCl3 and capsazepine and treatment with the inhibitors for CaMKII (Ca²2/calmodulin-dependent protein kinase II) and HSF1 (Heat shock factor 1) all significantly depressed the enhanced heat shock response (HSR). Together, we delineated the spatiotemporal dynamics of heat-induced calcium signaling and confirmed functions of the Ca2+-CaMKII-HSF1 pathway in regulating the HSR in zebrafish.
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影响因子：
7.2
作者：
Gilbert, Guillaume;Demydenko, Kateryna;Roderick, H. Llewelyn
通讯作者：
Roderick, H. Llewelyn
影响因子：
8.3
作者：
Christou, Maria;Fraser, Thomas W. K.;Kamstra, Jorke H.
通讯作者：
Kamstra, Jorke H.
脊椎动物心脏传导系统的遗传和生理解剖。
影响因子：
9.8
作者：
Chi NC;Shaw RM;Jungblut B;Huisken J;Ferrer T;Arnaout R;Scott I;Beis D;Xiao T;Baier H;Jan LY;Tristani-Firouzi M;Stainier DY
通讯作者：
Stainier DY
影响因子：
3.5
作者：
Kim CK;Miri A;Leung LC;Berndt A;Mourrain P;Tank DW;Burdine RD
通讯作者：
Burdine RD
DOI：
10.1038/nrm.2017.73
发表时间：
2018-01
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
Gomez-Pastor R;Burchfiel ET;Thiele DJ
通讯作者：
Thiele DJ