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改造和优化 GCaMP8f 使其可用于秀丽隐杆线虫。

Adapting and optimizing GCaMP8f for use in Caenorhabditis elegans.

机构信息

Max Planck Research Group Neural Information Flow, Max Planck Institute for Neurobiology of Behavior-caesar, Bonn 53175, Germany.

International Max Planck Research School for Brain and Behavior, Bonn 53175, Germany.

出版信息

Genetics. 2024 Oct 7;228(2). doi: 10.1093/genetics/iyae125.

DOI:10.1093/genetics/iyae125
PMID:39074213
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11457936/
Abstract

Improved genetically encoded calcium indicators (GECIs) are essential for capturing intracellular dynamics of both muscle and neurons. A novel set of GECIs with ultrafast kinetics and high sensitivity was recently reported by Zhang et al. (2023). While these indicators, called jGCaMP8, were demonstrated to work in Drosophila and mice, data for Caenorhabditis elegans were not reported. Here, we present an optimized construct for C. elegans and use this to generate several strains expressing GCaMP8f (fast variant of the indicator). Utilizing the myo-2 promoter, we compare pharyngeal muscle activity measured with GCaMP7f and GCaMP8f and find that GCaMP8f is brighter upon binding to calcium, shows faster kinetics, and is not disruptive to the intrinsic contraction dynamics of the pharynx. Additionally, we validate its application for detecting neuronal activity in touch receptor neurons which reveals robust calcium transients even at small stimulus amplitudes. As such, we establish GCaMP8f as a potent tool for C. elegans research which is capable of extracting fast calcium dynamics at very low magnifications across multiple cell types.

摘要

新型的超快动力学和高灵敏度的基因编码钙指示剂(GECIs)最近由 Zhang 等人报道。虽然这些被称为 jGCaMP8 的指示剂已被证明可在果蝇和小鼠中发挥作用,但尚未报告其在秀丽隐杆线虫中的数据。在这里,我们为秀丽隐杆线虫提供了一个经过优化的构建体,并利用它生成了几种表达 GCaMP8f(该指示剂的快速变体)的菌株。利用肌球蛋白-2 启动子,我们比较了用 GCaMP7f 和 GCaMP8f 测量的咽肌活性,发现 GCaMP8f 与钙结合时更亮,动力学更快,并且不会破坏咽的内在收缩动力学。此外,我们验证了其在检测触须感觉神经元中的神经元活性的应用,即使在小刺激幅度下也能产生强烈的钙瞬变。因此,我们确立了 GCaMP8f 作为秀丽隐杆线虫研究的有力工具,它能够在多个细胞类型中以非常低的放大倍数提取快速钙动力学。

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Adapting and optimizing GCaMP8f for use in Caenorhabditis elegans.改造和优化 GCaMP8f 使其可用于秀丽隐杆线虫。
Genetics. 2024 Oct 7;228(2). doi: 10.1093/genetics/iyae125.
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Brain-wide representations of behavior spanning multiple timescales and states in C. elegans.秀丽隐杆线虫中跨越多个时间尺度和状态的行为的全脑表达。
Cell. 2023 Sep 14;186(19):4134-4151.e31. doi: 10.1016/j.cell.2023.07.035. Epub 2023 Aug 21.
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Principles for coding associative memories in a compact neural network.在紧凑神经网络中对联想记忆进行编码的原则。
Elife. 2023 May 4;12:e74434. doi: 10.7554/eLife.74434.
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Fast and sensitive GCaMP calcium indicators for imaging neural populations.快速灵敏的 GCaMP 钙指示剂用于神经群体成像。
Nature. 2023 Mar;615(7954):884-891. doi: 10.1038/s41586-023-05828-9. Epub 2023 Mar 15.
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Automatically tracking feeding behavior in populations of foraging .自动跟踪觅食群体的进食行为。
Elife. 2022 Sep 9;11:e77252. doi: 10.7554/eLife.77252.
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Emerging imaging methods to study whole-brain function in rodent models.新兴的成像方法研究啮齿动物模型全脑功能。
Transl Psychiatry. 2021 Sep 4;11(1):457. doi: 10.1038/s41398-021-01575-5.
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Decoding locomotion from population neural activity in moving .从运动中的群体神经活动中解码运动。
Elife. 2021 Jul 29;10:e66135. doi: 10.7554/eLife.66135.
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Using Genetically Encoded Calcium Indicators to Study Astrocyte Physiology: A Field Guide.使用基因编码钙指示剂研究星形胶质细胞生理学:实用指南。
Front Cell Neurosci. 2021 Jun 11;15:690147. doi: 10.3389/fncel.2021.690147. eCollection 2021.
8
NeuroPAL: A Multicolor Atlas for Whole-Brain Neuronal Identification in C. elegans.NeuroPAL:秀丽隐杆线虫全脑神经元识别的多色图谱
Cell. 2021 Jan 7;184(1):272-288.e11. doi: 10.1016/j.cell.2020.12.012. Epub 2020 Dec 29.
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GABAergic motor neurons bias locomotor decision-making in C. elegans.GABA 能运动神经元影响秀丽隐杆线虫的运动决策。
Nat Commun. 2020 Oct 8;11(1):5076. doi: 10.1038/s41467-020-18893-9.
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Mapping Calcium Dynamics in the Heart of Zebrafish Embryos with Ratiometric Genetically Encoded Calcium Indicators.利用比率型基因编码钙指示剂绘制斑马鱼胚胎心脏中的钙动力学图谱。
Int J Mol Sci. 2020 Sep 10;21(18):6610. doi: 10.3390/ijms21186610.