Center for Cognition and Sociality, Institute for Basic Science (IBS), Daejeon, 34126, Republic of Korea.
Department of Bioscience and Biotechnology, Graduate School, Chungnam National University, Daejeon, 34134, Korea.
Mol Brain. 2023 Oct 17;16(1):73. doi: 10.1186/s13041-023-01061-7.
Calcium ions (Ca) 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 Ca-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 Ca-release-activated Ca (CRAC) channels in the plasma membrane through blue light, allowing precise control over intracellular Ca 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 Ca-mediated cellular activities in various brain functions. Furthermore, this toolkit holds potential as a therapeutic strategy for addressing brain disorders associated with aberrant Ca signaling.
钙离子 (Ca) 在调节多种大脑功能中起着关键作用，包括认知、情绪、运动、学习和记忆。这些功能是通过各种依赖 Ca 的细胞过程来精细调节的，包括突触可塑性、神经递质/胶质递质释放和基因表达。在我们之前的工作中，我们开发了“怪物 OptoSTIM1”（monSTIM1），这是一种经过改良的 OptoSTIM1，它可以通过蓝光选择性地激活质膜中的 Ca 释放激活的 Ca（CRAC）通道，从而可以精确控制细胞内 Ca 信号和特定的大脑功能。然而，monSTIM1 的编码序列较大，限制了其广泛应用，因为它超过了腺相关病毒（AAV）的包装能力。为了解决这个限制，我们引入了编码序列较小的 monSTIM1 变体，并建立了基于 AAV 的系统，用于在小鼠大脑中的神经元和神经胶质细胞中表达它们。通过 AAV 表达，这些 monSTIM1 变体在非侵入性光照射后显著增加了海马 CA1 区神经元和星形胶质细胞中 cFos 的表达水平。monSTIM1 变体的使用为研究 Ca 介导的细胞活动在各种大脑功能中的时空作用提供了一个有前途的途径。此外，该工具包作为一种治疗策略，具有解决与异常 Ca 信号相关的大脑疾病的潜力。
