Transcriptional Profile of Brain Endothelial Cells in Response to Perturbations of Glutamatergic Neural Acticity

Bulk RNA sequencing of Brain Endothelial Cells from brain regions that underwent 3 hours of DREADDs-mediated glutamatergic activation or silencing compared to respective littermate controls or volitional whisker-mediated barrel cortex activation compared to no whisker controls. Glutamatergic DREADDs mice and littermate controls that did not express DREADDs were injected with CNO to either activate or silence glutamatergic neural activity, then 3 hours later, cortices and hippocampi were dissected out, dissociated into a single cell suspension, live stained for CD31 and non-endothelial markers, CD31+ cells were enriched using FACS sorting, sequenced and compared to littermate controls. Gene expression profiles of 4 biological samples for each of the two DREADDs mice were sequenced and compared to their respesctive littermate controls. For the whisker experiments, wildtype mice explored a novelly enriched environment in the dark using their whiskers through voltional behavior, and then their barrel cortices were pooled together and the same brain enodthelial enrichment protocol was used. Gene expression profiles of 3 biological samples consisting of 5 pairs of barrel cortices were generated per group and compared to 3 biological samples consisting of 5 pairs of barrel cortices from wildtype mice without whiskers in an empty cage.