Identification and three-dimensional reconstruction of oxytocin receptor expressing astrocytes in the rat and mouse brain

Ferdinand Althammer; Eric G. Krause; Anette D. de Kloet; Justin Smith; Valery Grinevich; Alexandre Charlet; Javier E. Stern

STAR Protocols (Mar 2022)

Identification and three-dimensional reconstruction of oxytocin receptor expressing astrocytes in the rat and mouse brain

Ferdinand Althammer,
Eric G. Krause,
Anette D. de Kloet,
Justin Smith,
Valery Grinevich,
Alexandre Charlet,
Javier E. Stern

Affiliations

Ferdinand Althammer: Center for Neuroinflammation and Cardiometabolic Diseases, Georgia State University, Atlanta, GA 30302-5030, USA; Corresponding author
Eric G. Krause: Department of Pharmacodynamics, College of Pharmacy, University of Florida, Gainesville, FL 32611, USA; Center of Integrative Cardiovascular and Metabolic Diseases, University of Florida, Gainesville, FL 32611, USA; McKnight Brain Institute, University of Florida, Gainesville, FL 32611, USA
Anette D. de Kloet: Department of Physiology and Functional Genomics, College of Medicine, University of Florida, Gainesville, FL 32611, USA; Center of Integrative Cardiovascular and Metabolic Diseases, University of Florida, Gainesville, FL 32611, USA; McKnight Brain Institute, University of Florida, Gainesville, FL 32611, USA
Justin Smith: Department of Pharmacodynamics, College of Pharmacy, University of Florida, Gainesville, FL 32611, USA; Center of Integrative Cardiovascular and Metabolic Diseases, University of Florida, Gainesville, FL 32611, USA; McKnight Brain Institute, University of Florida, Gainesville, FL 32611, USA
Valery Grinevich: Center for Neuroinflammation and Cardiometabolic Diseases, Georgia State University, Atlanta, GA 30302-5030, USA; Department of Neuropeptide Research for Psychiatry, Central Institute of Mental Health, University of Heidelberg, Mannheim, Germany
Alexandre Charlet: Centre National de la Recherche Scientifique, University of Strasbourg, Strasbourg, France; Institute of Cellular and Integrative Neuroscience, INCI CNRS UPR3212 8, Allée du Général Rouvillois 67000 Strasbourg, France; Corresponding author
Javier E. Stern: Center for Neuroinflammation and Cardiometabolic Diseases, Georgia State University, Atlanta, GA 30302-5030, USA; Corresponding author

Journal volume & issue: Vol. 3, no. 1
p. 101160

Abstract

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Summary: Here, we present a step-by-step protocol for three-dimensional reconstruction of astrocyte morphology, applied to the central amygdala oxytocin receptor-expressing astrocytes. This includes RNAse-free perfusion, combination of RNAscope and immunohistochemistry, and confocal imaging. This protocol provides detailed information about tissue handling and a comprehensive description of the RNAScope technique to label rat and mouse oxytocin receptor mRNA. We also describe three-dimensional reconstruction that allows the assessment of more than 70 different cellular parameters, powerful for studying astrocyte morphology and astrocyte-astrocyte interactions.For complete details on the use and execution of this protocol, please refer to Wahis et al. (2021) and Althammer et al. (2020).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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