Use of Cre-Lox Technology to Analyze Integrin Functions in Astrocytes

Astrocytes communicate with the vascular endothelium via direct cell–cell contacts as well as a variety of secreted growth factors and extracellular matrix (ECM) proteins. Integrins are heterodimeric cell surface receptors for ECM protein ligands, and many integrin subunits are expressed in astrocytes. Here, we will discuss gene deletion strategies in mice that have deciphered functions for specific integrins in astrocyte-endothelial cell adhesion and signaling. Specifically, we will detail how Cre-lox molecular genetic techniques have revealed important roles for integrin αvβ8 in regulating cerebral blood vessel development and homeostasis. First, we will detail how to generate Cre-lox mutant mouse models that our group and others have used to study αvβ8 integrin in embryonic astroglial progenitors and postnatal astrocytes. Second, we will discuss how viral-delivered Cre can be used to acutely delete integrin genes in astrocytes within defined anatomic regions of the brain. Third, detailed in vivo methods to verify Cre-mediated gene recombination in astrocytes will be presented. Lastly, we will present one experimental strategy to determine how integrin gene deletion affects astrocyte-endothelial cell coupling in the CNS. While this review focuses on the generation and characterization of mice lacking αvβ8 integrin, these experimental strategies can be expanded to analyze other cell adhesion and signaling genes important for astroglial-mediated regulation of blood vessel development and homeostasis.