Revealing and Characterizing Astrocytic Atrophy in an Animal Model of Depression

Astrocyte atrophy is the main histopathological hallmark of major depressive disorder (MDD) in humans and in animal models of depression. In particular, depression and depressive-like behaviors are associated with a substantial decrease in size and complexity of astrocytes in the prefrontal cortex. This results in the reduced homeostatic support of synaptic transmission, which arguably translates into abnormal activity of neuronal networks and pathological changes in mood and behavior. Treatment of experimental animals with antidepressants as well as with acupuncture in the specific acupoints alleviates depressive-like behaviors and rescues astrocytic atrophy. Here, we describe the methodology for inducing and monitoring the depressive-like behaviors in mice using chronic unpredictable mild stress (CUMS), with subsequent in-depth analysis of astrocytic morphology using confocal microscopy in conjunction with astrocyte-specific labeling by virally transfected genetically encoded fluorescent probe mCherry and intracellular injection of Lucifer yellow. We also describe immunocytochemical visualization of astrocyte-specific cytoskeletal linker ezrin, which is involved in controlling perisynaptic astrocytic leaflets.