An Improved Method for Physical Separation of Cerebral Vasculature and Parenchyma Enables Detection of Blood-Brain-Barrier Dysfunction

The neurovascular niche is crucial for constant blood supply and blood-brain barrier (BBB) function altered in a number of different neurological conditions, making this an intensely active field research. Brain vasculature unique its tight association endothelial cells with astrocytic endfeet processes. Separation the vascular compartment by centrifugation-based methods confirmed enrichment processes, it possible to study entire such methods. Several separation protocols are found literature; however, some constraints which limit their applicability scope studies. Here, we describe validate protocol physically separating from parenchyma, optimized smaller tissue quantities. Using endothelial, neuronal, astrocyte markers, show that quantitative Western blot-based target detection can be performed both vessel-enriched parenchymal fractions using as little single mouse brain hemisphere. Validation our rodent stroke models detecting changes junction protein expression, serum albumin signals activation, i.e., increased glial fibrillary acidic between ipsilateral lesion-free contralateral hemisphere demonstrates new way BBB breakdown astrogliosis, respectively.