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Summary: The present investigation was carried out to determine the extent to which muscarinic acetylcholine receptors (mAChRs) in vascular and perivascular struc tures were colocalized with glial fibrillary acidic protein (GFAP)-positive structures. To this aim, an immunocy tochemical approach on free-floating cryosections and isolated microvessels obtained from rat brain was per formed to study the possible colocalization of immuno staining with the anti-mAChR protein antibody (M35) and an anti-GF AP antibody. Double-labeling experiments were carried out by fluorescent techniques. Confocal mi croscopic observations of GF AP and M35 immunoreac tivities on free-floating sections showed a high degree of colocalization on astrocyte processes associated with large vessels or capillaries. This pattern suggests that muscarinic receptors are associated with astrocytic end feet. Confocal microscopic observations of immunoreac tivity from isolated cerebral microvessels strengthen this conclusion since double-labeling of M35 and GFAP showed that perivascular astrocytic structures remained

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