Adapting and optimizing GCaMP8f for use in Caenorhabditis elegans.

IF 3.3 3区生物学 Q2 GENETICS & HEREDITY

Genetics Pub Date : 2024-10-07 DOI:10.1093/genetics/iyae125

Jun Liu, Elsa Bonnard, Monika Scholz

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引用次数: 0

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.

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调整和优化 GCaMP8f，使其用于秀丽隐杆线虫。

改进的基因编码钙离子指示剂（GECIs）对于捕捉肌肉和神经元的细胞内动态至关重要。Zhang 等人最近报道了一组具有超快动力学和高灵敏度的新型 GECIs（《自然》，2023 年）。这些被称为 jGCaMP8 的指标已在果蝇和小鼠中证实有效，但在秀丽隐杆线虫中的数据却未见报道。在这里，我们提出了一种针对秀丽隐杆线虫的优化构建物，并利用它生成了几种表达 GCaMP8f（指示剂的快速变体）的品系。利用 myo-2 启动子，我们比较了用 GCaMP7f 和 GCaMP8f 测得的咽部肌肉活动，发现 GCaMP8f 在与钙结合时更亮，显示出更快的动力学，而且不会破坏咽部的内在收缩动力学。此外，我们还验证了 GCaMP8f 在检测触觉受体神经元活动中的应用，即使在刺激幅度较小的情况下也能显示出强大的钙瞬态。因此，我们将 GCaMP8f 确立为 elegans 研究的有效工具，它能够以极低的放大倍数提取多种细胞类型的快速钙动态。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Genetics GENETICS & HEREDITY-

CiteScore

6.90

自引率

6.10%

发文量

177

审稿时长

1.5 months

期刊介绍： GENETICS is published by the Genetics Society of America, a scholarly society that seeks to deepen our understanding of the living world by advancing our understanding of genetics. Since 1916, GENETICS has published high-quality, original research presenting novel findings bearing on genetics and genomics. The journal publishes empirical studies of organisms ranging from microbes to humans, as well as theoretical work. While it has an illustrious history, GENETICS has changed along with the communities it serves: it is not your mentor''s journal. The editors make decisions quickly – in around 30 days – without sacrificing the excellence and scholarship for which the journal has long been known. GENETICS is a peer reviewed, peer-edited journal, with an international reach and increasing visibility and impact. All editorial decisions are made through collaboration of at least two editors who are practicing scientists. GENETICS is constantly innovating: expanded types of content include Reviews, Commentary (current issues of interest to geneticists), Perspectives (historical), Primers (to introduce primary literature into the classroom), Toolbox Reviews, plus YeastBook, FlyBook, and WormBook (coming spring 2016). For particularly time-sensitive results, we publish Communications. As part of our mission to serve our communities, we''ve published thematic collections, including Genomic Selection, Multiparental Populations, Mouse Collaborative Cross, and the Genetics of Sex.