human brain capillary endothelial cells  (Cedarlane)

Journal: Nature Communications

Article Title: A circadian clock regulates efflux by the blood-brain barrier in mice and human cells

doi: 10.1038/s41467-020-20795-9

Figure Lengend Snippet: RHB was intravenously injected via jugular vein into mice. Mice were allowed 90 min to recover and brains and sera were collected. Fluorescence was read at ex540/em590nm using a plate reader. a Clock ablation in endothelial cells disrupts the brain permeability rhythm to RHB. RHB was injected into control ( n = 18; 4 independent experiments) or endothelial-specific Bmal1 -deficient ( n = 31; 4 independent experiments) mice at indicated time points. b Global circadian disruption ablates brain permeability rhythm to RHB in mice. RHB was injected into Cry1/2 het ( n = 24; 3 independent experiments) and Cry1/2 DKO ( n = 23; 4 independent experiments) mice at indicated time points. Individual mice are shown with triangle markers, and lines represent the mean. pCycle values were calculated by JTKCycle analysis.

Article Snippet: Immortalized human brain capillary endothelial cells (Millipore SCC066) were cultured in EndoGRO-MV Complete media kit (SCME004) as recommended by the manufacturer.

Techniques: Injection, Fluorescence, Permeability, Control, Disruption

Journal: Nature Communications

Article Title: A circadian clock regulates efflux by the blood-brain barrier in mice and human cells

doi: 10.1038/s41467-020-20795-9

Figure Lengend Snippet: Control or endothelial-cell specific Bmal1-deficient mice were collected at ZT2, ZT6, ZT10, ZT14, ZT18, and ZT22 ( n = 12; 6 time points, 2 independent experiments); brains were dissected and endothelial cells were isolated by FACS using CD31 antibody. RNA was extracted and sequenced with HiSeq. a Few transcripts cycle in BBB endothelial cells. Heatmap of cycling transcripts of control animals (Meta2d, q < 0.6) is organized by peak and trough with corresponding transcripts of Bmal1 -deficient endothelial cells. b Expression of core circadian genes shows robust rhythms in mouse BBB. Transcripts of controls (black) are rhythmic (pCycle <0.001) while Bmal1 -deficient endothelial cells (red) are not. pCycle values were calculated by Meta2d analysis adjusted for multiple comparisons. c Trpm7 is highly expressed in BBB endothelium. Expression of known magnesium transporters in controls ( n = 12; 2 independent experiments) are shown as means ± SEM. d Trpm7 shows rhythmic expression in brain endothelial cells. qPCR of sorted brain endothelial cells ( n = 24; 6 time points, 4 independent experiments) were probed for Nr1d1 and Trpm7 expression across the circadian day normalized to GAPDH . pCycle values were calculated by JTKCycle analysis.

Article Snippet: Immortalized human brain capillary endothelial cells (Millipore SCC066) were cultured in EndoGRO-MV Complete media kit (SCME004) as recommended by the manufacturer.

Techniques: Control, Isolation, Expressing