Chemogenetic Silencing of Neonatal Spontaneous Activity of Projection Neurons in the Dorsal Striatum of Mice.

IF 1 Q3 BIOLOGY Bio-protocol Pub Date : 2024-10-20 DOI:10.21769/BioProtoc.5088
Bojana Kokinovic, Maria Ryazantseva, Svetlana Molchanova
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引用次数: 0

Abstract

Neuroscience incorporates manipulating neuronal circuitry to enhance the understanding of intricate brain functions. An effective strategy to attain this objective entails utilizing viral vectors to induce varied gene expression by delivering transgenes into brain cells. Here, we combine the use of transgenic mice, neonatal transduction with adeno-associated viral constructs harboring inhibitory designer receptor exclusively activated by designer drug (DREADD) gene, and the DREADD agonist clozapine N-oxide (CNO). In this way, a chemogenetic approach is employed to suppress neuronal activity in the region of interest during a critical developmental window, with subsequent investigation into its effects on the neuronal circuitry in adulthood. Key features • Comprehensive protocol for newborn viral transduction in the dorsal striatum of mice • Uses a viral construct encoding inhibitory DREADD under the control of Cre recombinase to attenuate the activity of specific cell types in the brain.

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化学基因沉默小鼠背侧纹状体投射神经元的新生儿自发活动
神经科学通过操纵神经元回路来加深对复杂大脑功能的理解。实现这一目标的有效策略是利用病毒载体将转基因送入脑细胞,诱导不同的基因表达。在这里,我们结合使用转基因小鼠、新生儿转导腺相关病毒构建体(携带抑制性设计药物受体(DREADD)基因)和 DREADD 激动剂氯氮平 N-氧化物(CNO)。通过这种方法,可以利用化学遗传学方法在关键的发育窗口期抑制相关区域的神经元活动,并随后研究其对成年期神经元回路的影响。主要特点 - 在小鼠背侧纹状体中进行新生儿病毒转导的综合方案 - 在 Cre 重组酶的控制下使用编码抑制性 DREADD 的病毒构建体来削弱大脑中特定细胞类型的活性。
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