Affiliation:
1. Queen Mary School Nanchang University Nanchang China
2. Laboratory of Fear and Anxiety Disorders, Institutes of Life Science Nanchang University Nanchang China
3. Department of Biological Science, School of Life Science Nanchang University Nanchang China
4. Center for Medical Experiments Nanchang University Nanchang China
Abstract
AbstractOne of the most sought‐after topics in neuroscience is to understand how the environment regulates the activity and function of neural circuitry and subsequently influences relevant behaviors. In response to alterations in the environment, the neural circuits undergo adaptive changes ranging from gene expression changes to altered cellular function. Performing sequencing of the transcriptome involved in these behavior‐related circuits will provide clues to accurately dissect the detailed mechanisms of related behavior. Here, we describe methods for marking and collecting the ventral hippocampus–projecting basolateral amygdala neurons, which have been repeatedly implicated in regulation of anxiety‐like behavior, and subsequently constructing a library ready for sequencing. Specifically, the reported approaches include adeno‐associated virus injection, acute brain slice isolation, cell suspension preparation, cell extraction, and cDNA library construction. By utilizing the techniques described here, researchers can comprehensively investigate the transcriptional levels of neural clusters embedded in particular circuits and discover potential pathogenic and therapeutic targets for behavior‐relevant disorders. © 2023 Wiley Periodicals LLC.Basic Protocol 1: Tagging of behavior‐related neural circuitsBasic Protocol 2: Isolation and capture of fluorescent‐positive cellsBasic Protocol 3: Foundation of sequencing library
Subject
Medical Laboratory Technology,Health Informatics,General Pharmacology, Toxicology and Pharmaceutics,General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Neuroscience