Examining the Coverage of Blood Vessels by Astrocyte Endfeet in an Animal Model of Major Depressive Disorder
The use of immunofluorescent immunohistochemical methods has been instrumental to characterize the distribution and extents of colocalization of molecules located in various cellular compartments. Such information has been pivotal to formulate hypothesis about their roles in physiological and pathological conditions.
In the brain, astrocytes interact with endothelial cells to form the blood–brain barrier (BBB). Astrocytes regulate features of the BBB through the tips of their processes, called astrocyte endfeet, which surround and contact brain microvessels. Any disruption in astrocyte endfeet can have dramatic consequences on the integrity of the BBB, with consequent development of brain disorders.
Here we describe a method to characterize the integrity of astrocyte endfeet around blood vessels in an animal model of major depressive disorder (MDD) using confocal microscopy, Adobe Photoshop, and the AutoQuantX3 software.
Key wordsLaser scanning microscopy Astrocytes Perivascular endfeet Prefrontal cortex Major depressive disorder Blood–brain barrier
The authors wish to thank Salina Begum and Dr. Victoria Malik for the execution of some experiments used to prepare the images for the protocol presented here. This work was supported by intramural funding from the University of Regensburg, by the German Federal Ministry of Education and Research (BMBF Grant Nr 01EE1401A), and by the German Research Council (DFG GRK2174) to BDB.
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