RT Journal Article SR Electronic T1 Universal method for the gentle isolation of intact microvessels from frozen tissue: a multiomic investigation into the neurovasculature JF bioRxiv FD Cold Spring Harbor Laboratory SP 2023.05.10.540076 DO 10.1101/2023.05.10.540076 A1 Wakid, Marina A1 Almeida, Daniel A1 Aouabed, Zahia A1 Rahimian, Reza A1 Davoli, Maria Antonietta A1 Yerko, Volodymyr A1 Leonova-Erko, Elena A1 Richard, Vincent A1 Zahedi, René A1 Borchers, Christoph A1 Turecki, Gustavo A1 Mechawar, Naguib YR 2023 UL http://biorxiv.org/content/early/2023/05/11/2023.05.10.540076.1.abstract AB The neurovascular unit (NVU), comprised of endothelial cells, pericytes, smooth muscle cells, astrocytic endfeet and microglia together with neurons, is paramount for the proper function of the central nervous system. The NVU gatekeeps blood-brain barrier (BBB) properties which, as a system, experiences impairment in several neurological and psychiatric diseases, and contributes to pathogenesis. To better understand function and dysfunction at the NVU, isolation and characterization of the NVU is needed. Here, we describe a singular, standardized protocol to enrich and isolate microvessels from archived snap-frozen human and frozen mouse cerebral cortex using mechanical homogenization and centrifugation-separation that preserves the structural integrity and multicellular composition of microvessel fragments. For the first time, microvessels are isolated from postmortem vmPFC tissue and are comprehensively investigated using both RNA sequencing and Liquid Chromatography with tandem mass spectrometry (LC-MS-MS). Both the transcriptome and proteome are elucidated and compared, demonstrating that the isolated brain microvessel is a robust model for the NVU and can be used to generate highly informative datasets in both physiological and disease contexts.Competing Interest StatementThe authors have declared no competing interest.