Reducing astrocyte calcium signaling in vivo alters striatal microcircuits and causes repetitive behavior II. Reducing astrocyte calcium signaling in vivo alters striatal microcircuits and causes repetitive behavior II

Astrocytes tile the central nervous system, but their functions in neural microcircuits in vivo and their roles in mammalian behavior remain incompletely defined. We used 2-photon laser scanning microscopy (2PLSM), electrophysiology, MINIscopes, RNA-seq and a new genetic approach to characterize the effects of reduced striatal astrocyte Ca2+ signaling in vivo. In wild type mice, reducing striatal astrocyte Ca2+-dependent signaling increased repetitive self-grooming behaviors by altering medium spiny neuron (MSN) activity. The mechanism involved astrocyte-mediated neuromodulation mediated by ambient GABA and was corrected by blocking astrocyte GABA transporter 3 (GAT-3). Furthermore, in a mouse model of Huntington’s disease, dysregulation of GABA and astrocyte Ca2+ signaling accompanied excessive self-grooming, which was relieved by blocking GAT-3. Assessments with RNA-seq revealed astrocyte genes and pathways regulated by Ca2+ signaling in a cell autonomous and non-cell autonomous manner, including Rab11a, a regulator of GAT-3 functional expression. Thus, striatal astrocytes contribute to neuromodulation controlling obsessive-compulsive-like behavior in mice. Overall design: Total 16 samples were used in this data set. Whole striata were extraced from 8 adult Aldh1l1-Cre/ERT2 x Ribotag mice: 4 mice received AAV encoding a control protein and 4 mice received AAV encoding a protein that suppresses calcium signals named hPMCA2w/b. After homogenization, RNA was purified from (i) cleared lysate as the input and control, and (ii) astrocyte-specific ribosome-associated RNA precipitated via a hemagglutinin (HA) tag.

星形胶质细胞（Astrocytes）铺展覆盖中枢神经系统，但它们在体内神经微环路中的功能，以及在哺乳动物行为中的作用仍未被完全阐明。我们采用双光子激光扫描显微镜（2-photon laser scanning microscopy，2PLSM）、电生理学技术、MINIscopes微型钙成像系统、RNA测序（RNA-seq）以及一种新型遗传学方法，在活体状态下表征了纹状体星形胶质细胞钙信号减弱所产生的生物学效应。在野生型小鼠中，抑制纹状体星形胶质细胞的钙依赖信号通路，会通过改变中型多棘神经元（medium spiny neuron，MSN）的活动，增加重复性自我理毛行为。该机制涉及星形胶质细胞通过胞外γ-氨基丁酸（GABA）介导的神经调节，且可通过阻断星形胶质细胞γ-氨基丁酸转运体3（GABA transporter 3，GAT-3）得以纠正。此外，在亨廷顿病小鼠模型中，γ-氨基丁酸失衡与星形胶质细胞钙信号异常共同伴随出现过度自我理毛行为，而阻断GAT-3可缓解该症状。RNA测序分析显示，星形胶质细胞的基因与通路可通过钙信号以细胞自主性及非细胞自主性的方式进行调控，其中包括调控GAT-3功能性表达的Rab11a。综上，纹状体星形胶质细胞参与了调控小鼠类强迫样行为的神经调节过程。整体实验设计：本数据集共使用16个样本。从8只成年Aldh1l1-Cre/ERT2 x Ribotag小鼠中提取全纹状体组织：其中4只小鼠注射编码对照蛋白的腺相关病毒（AAV），另外4只小鼠注射编码可抑制钙信号的hPMCA2w/b蛋白的腺相关病毒。组织匀浆后，分别从（i）澄清裂解液中纯化RNA作为输入样本与对照，以及（ii）通过血凝素（hemagglutinin，HA）标签沉淀得到的星形胶质细胞特异性核糖体结合RNA。