Immunofluorescence Protocol to Label Astrocytes from Diverse Model and Non-Model Mammals -- Retro-Orbital Delivery of AAVs for CNS-Wide Astrocyte Targeting -- An Empirical-Theoretical Approach to Determine Astroglial Potassium upon Ischemic Stress -- Quantification of Astrocytic Sodium Signals Using Fluorescence Lifetime Imaging (FLIM) -- pH Sensor-Based Engulfment Assays to Measure Phagocytic Activity of Astrocytes -- An Astrocyte Transplantation Method to Investigate Astrocyte Development and Diversity in the CNS -- Primary CNS Cultures with Mixed Neural Cell Types to Study Correlative Effects of High Glucocorticoids on Astrocytes, Oligodendrocytes, and Myelination Markers -- Postnatal Development of Perivascular Astrocytic Processes and Detection of Local mRNA and Translation -- Imaging Techniques for the Analysis of the Blood-Brain Barrier and Gliovascular Unit Dysfunction -- Assessment of Blood-Brain Barrier Integrity Using Dyes and Tracers -- Investigating the Pathology-Related Astroglial Plasticity in the Human Cerebral Cortex -- Super-Resolution Analysis of Astrocyte Morphology Using Expansion Microscopy -- Astrocyte Morphological Adaptations in a Mouse Model of Social Anxiety Disorder -- An In Vitro Assay to Measure Astrocyte-Dependent Synaptic Phagocytosis in Health and Major Depressive Disorder -- Cortical Organotypic Brain Slice Cultures to Examine Sex- and Age-Dependent Astrocyte-Mediated Synaptic Phagocytosis -- Development and Validation of Stress Paradigms to Study Astrocytes in Rodent Models of Stress Disorders -- Cellular Investigation of Astrocytic Neuroendocrine Responses to Stress -- Morphological Investigation of Astrocytic Responses to Stress -- Functional Evaluation of Astrocytic Responses to Stress -- Unveiling Ketamine's Influence on Astrocytic Kir4.1 Channels through Multi-Modal Analysis: Confocal Microscopy, Immunocytochemistry, Fluorescence Analysis, and Electrophysiology -- Revealing and Characterizing Astrocytic Atrophy in an Animal Model of Depression.

This detailed volume gathers methods essential for investigating the multifaceted roles of astrocytes in both physiology and pathological conditions. The book covers several techniques for the preparation of astrocytes from animal models, the examination of astrocyte morphology and functions in vitro and ex vivo, as well as the investigation of astrocyte pathologies in the central nervous system. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Astrocytes: Methods and Protocols, Second Edition serves as an ideal guide for both experienced and beginner scientists working toward unravelling the roles of these highly versatile cells.