AAV-compatible optogenetic tools for activating endogenous calcium channels in vivo

Molecular Brain

Methodology

Hyoin Lee¹  Sangkyu Lee¹  Yeon Hee Kook²  Jinsu Lee³  Yeonji Jeong³  Won Do Heo⁴  Jaerang Rho⁵ 

[1] Center for Cognition and Sociality, Institute for Basic Science (IBS), 34126, Daejeon, Republic of Korea;Center for Cognition and Sociality, Institute for Basic Science (IBS), 34126, Daejeon, Republic of Korea;Department of Bioscience and Biotechnology, Graduate School, Chungnam National University, 34134, Daejeon, Korea;Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), 34141, Daejeon, Republic of Korea;Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), 34141, Daejeon, Republic of Korea;KAIST Institute for the BioCentury, Korea Advanced Institute of Science and Technology (KAIST), 34141, Daejeon, Republic of Korea;Department of Bioscience and Biotechnology, Graduate School, Chungnam National University, 34134, Daejeon, Korea;

关键词: Calcium ion;    Optogenetics;    Adeno-associated virus;    Neurons;    Glial cells;   

DOI  :  10.1186/s13041-023-01061-7

 received in 2023-08-30, accepted in 2023-09-28,  发布年份 2023

来源: Springer

【 摘 要 】

Calcium ions (Ca2+) play pivotal roles in regulating diverse brain functions, including cognition, emotion, locomotion, and learning and memory. These functions are intricately regulated by a variety of Ca2+-dependent cellular processes, encompassing synaptic plasticity, neuro/gliotransmitter release, and gene expression. In our previous work, we developed ‘monster OptoSTIM1’ (monSTIM1), an improved OptoSTIM1 that selectively activates Ca2+-release–activated Ca2+ (CRAC) channels in the plasma membrane through blue light, allowing precise control over intracellular Ca2+ signaling and specific brain functions. However, the large size of the coding sequence of monSTIM1 poses a limitation for its widespread use, as it exceeds the packaging capacity of adeno-associated virus (AAV). To address this constraint, we have introduced monSTIM1 variants with reduced coding sequence sizes and established AAV-based systems for expressing them in neurons and glial cells in the mouse brain. Upon expression by AAVs, these monSTIM1 variants significantly increased the expression levels of cFos in neurons and astrocytes in the hippocampal CA1 region following non-invasive light illumination. The use of monSTIM1 variants offers a promising avenue for investigating the spatiotemporal roles of Ca2+-mediated cellular activities in various brain functions. Furthermore, this toolkit holds potential as a therapeutic strategy for addressing brain disorders associated with aberrant Ca2+ signaling.

【 授权许可】

CC BY   
© Min Zhuo, Bong-Kiun Kaang and BioMed central Ltd. 2023

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