A sensitive soma-localized red fluorescent calcium indicator for in vivo imaging of neuronal populations at single-cell resolution.

IF 9.8 1区 生物学 Q1 Agricultural and Biological Sciences
PLoS Biology Pub Date : 2025-04-29 eCollection Date: 2025-04-01 DOI:10.1371/journal.pbio.3003048
Shihao Zhou, Qiyu Zhu, Minho Eom, Shilin Fang, Oksana M Subach, Chen Ran, Jonnathan Singh Alvarado, Praneel S Sunkavalli, Yuanping Dong, Yangdong Wang, Jiewen Hu, Hanbin Zhang, Zhiyuan Wang, Xiaoting Sun, Tao Yang, Yu Mu, Young-Gyu Yoon, Zengcai V Guo, Fedor V Subach, Kiryl D Piatkevich
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引用次数: 0

Abstract

Recent advancements in genetically encoded calcium indicators, particularly those based on green fluorescent proteins, have optimized their performance for monitoring neuronal activities in a variety of model organisms. However, progress in developing red-shifted GECIs, despite their advantages over green indicators, has been slower, resulting in fewer options for end users. In this study, we explored topological inversion and soma-targeting strategies, which are complementary to conventional mutagenesis, to re-engineer a red genetically encoded calcium indicator, FRCaMP, for enhanced in vivo performance. The resulting sensors, FRCaMPi and soma-targeted FRCaMPi (SomaFRCaMPi), exhibit up to 2-fold higher dynamic range and peak ΔF/F0 per single AP compared to widely used jRGECO1a in neurons both in culture and in vivo. Compared to jRGECO1a and FRCaMPi, SomaFRCaMPi reduces erroneous correlation of neuronal activity in the brains of mice and zebrafish by two- to 4-fold due to diminished neuropil contamination without compromising the signal-to-noise ratio. Under wide-field imaging in primary somatosensory and visual cortices in mice with high labeling density (80-90%), SomaFRCaMPi exhibits up to 40% higher SNR and decreased artifactual correlation across neurons. Altogether, SomaFRCaMPi improves the accuracy and scale of neuronal activity imaging at single-neuron resolution in densely labeled brain tissues due to a 2-3-fold enhanced automated neuronal segmentation, 50% higher fraction of responsive cells, up to 2-fold higher SNR compared to jRGECO1a. Our findings highlight the potential of SomaFRCaMPi, comparable to the most sensitive soma-targeted GCaMP, for precise spatial recording of neuronal populations using popular imaging modalities in model organisms such as zebrafish and mice.

一种敏感的体细胞定位红色荧光钙指示剂,用于单细胞分辨率下神经元群体的体内成像。
遗传编码钙指标的最新进展,特别是那些基于绿色荧光蛋白的钙指标,已经优化了它们在各种模式生物中监测神经元活动的性能。然而,开发红移geci的进展尽管比绿色指标有优势,但速度较慢,导致最终用户的选择较少。在这项研究中,我们探索了拓扑倒置和体细胞靶向策略,这些策略是传统诱变的补充,以重新设计红色遗传编码钙指示剂FRCaMP,以提高体内性能。由此产生的传感器,FRCaMPi和SomaFRCaMPi (SomaFRCaMPi),在培养和体内的神经元中,与广泛使用的jRGECO1a相比,表现出高达2倍的动态范围和峰值ΔF/F0。与jRGECO1a和FRCaMPi相比,SomaFRCaMPi减少了小鼠和斑马鱼大脑中神经元活动的错误相关性,减少了2至4倍,因为神经污染减少了,而不影响信噪比。在高标记密度(80-90%)小鼠初级体感觉和视觉皮层的宽视场成像下,SomaFRCaMPi显示出高达40%的高信噪比,并降低了神经元之间的人工相关性。总的来说,SomaFRCaMPi提高了在密集标记的脑组织中以单神经元分辨率进行神经元活动成像的准确性和规模,因为与jRGECO1a相比,SomaFRCaMPi的自动神经元分割增强了2-3倍,反应细胞的比例提高了50%,信噪比提高了2倍。我们的研究结果强调了SomaFRCaMPi的潜力,可以与最敏感的soma靶向GCaMP相媲美,在斑马鱼和小鼠等模式生物中使用流行的成像方式对神经元种群进行精确的空间记录。
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来源期刊
PLoS Biology
PLoS Biology BIOCHEMISTRY & MOLECULAR BIOLOGY-BIOLOGY
CiteScore
15.40
自引率
2.00%
发文量
359
审稿时长
3-8 weeks
期刊介绍: PLOS Biology is the flagship journal of the Public Library of Science (PLOS) and focuses on publishing groundbreaking and relevant research in all areas of biological science. The journal features works at various scales, ranging from molecules to ecosystems, and also encourages interdisciplinary studies. PLOS Biology publishes articles that demonstrate exceptional significance, originality, and relevance, with a high standard of scientific rigor in methodology, reporting, and conclusions. The journal aims to advance science and serve the research community by transforming research communication to align with the research process. It offers evolving article types and policies that empower authors to share the complete story behind their scientific findings with a diverse global audience of researchers, educators, policymakers, patient advocacy groups, and the general public. PLOS Biology, along with other PLOS journals, is widely indexed by major services such as Crossref, Dimensions, DOAJ, Google Scholar, PubMed, PubMed Central, Scopus, and Web of Science. Additionally, PLOS Biology is indexed by various other services including AGRICOLA, Biological Abstracts, BIOSYS Previews, CABI CAB Abstracts, CABI Global Health, CAPES, CAS, CNKI, Embase, Journal Guide, MEDLINE, and Zoological Record, ensuring that the research content is easily accessible and discoverable by a wide range of audiences.
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