PT  - JOURNAL ARTICLE
AU  - Wakid, Marina
AU  - Almeida, Daniel
AU  - Aouabed, Zahia
AU  - Rahimian, Reza
AU  - Davoli, Maria Antonietta
AU  - Yerko, Volodymyr
AU  - Leonova-Erko, Elena
AU  - Richard, Vincent
AU  - Zahedi, René
AU  - Borchers, Christoph
AU  - Turecki, Gustavo
AU  - Mechawar, Naguib
TI  - Universal method for the gentle isolation of intact microvessels from frozen tissue: a multiomic investigation into the neurovasculature
AID  - 10.1101/2023.05.10.540076
DP  - 2023 Jan 01
TA  - bioRxiv
PG  - 2023.05.10.540076
4099  - http://biorxiv.org/content/early/2023/05/11/2023.05.10.540076.1.short
4100  - http://biorxiv.org/content/early/2023/05/11/2023.05.10.540076.1.full
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.