Endothelium-derived lactate is required for pericyte function and blood-brain barrier maintenance (2022)
Figure 1

Figure 1. Loss of GLUT1 in EC decreases pericyte coverage on the vasculature and alters BBB permeability.

A. Retinal whole mount immunostaining for Isolectin-B4 (red) and Desmin (green) in WT (top panel) and Glut1iECKO (bottom panel) mice showing pericyte coverage in the vasculature. Left and right panels show representative images with 20x or 63x magnification, respectively.

B. Quantification of the pericytes coverage in the retinal vasculature of WT (blue) and Glut1iECKO (orange) mice. (n=9 from 3 independent experiments. Error bars indicate the standard error of the mean (SEM) from unpaired Student's t test.)

C. Immunostaining for CD31 (green), CD13 (white; pericyte marker) and αSMA (cyan) using paraffin section from WT (top panel) and Glut1iECKO (bottom panel) mice.

D. Quantification of the pericytes coverage using CD13 immunostaining in the brain vasculature of WT (blue) and Glut1iECKO (orange) mice. (n=7 from 3 independent experiments. Error bars indicate the standard error of the mean (SEM) from unpaired Student's t test.)

E. Quantification of the smooth muscle cell coverage using αSMA immunostaining in the brain vasculature of WT (blue) and Glut1iECKO (orange) mice. (n=13 from 3 independent experiments. Error bars indicate the standard error of the mean (SEM) from unpaired Student's t test.)

F. Immunostaining for Cldn5 (green) and CD31 (red) using a brain section of WT (top panel) and Glut1iECKO (bottom panel) mice.

G. Immunostaining for caspas-3 (green), CD13 (red) and CD31 (cyan) using a brain section of WT (top panel) and Glut1iECKO (bottom panel) mice.

H. Permeability assay using vibratome section showing the leakage of retro-orbital injected cadaverine dye (red) in the brain of WT (top panel) and Glut1iECKO (bottom panel) mice. DAPI staining (white) shows the boundary of brain section.

I. Permeability assay using vibratome section showing the leakage of retro-orbital injected cadaverine dye (red) in the brain of WT (top panel) and Glut1iECKO (bottom panel) mice. Note that cadaverine is retained in the intravascular area of WT mice (upper panel), but extravasated and accumulated in the brain parenchyma (white arrowheads) of Glut1iECKO (bottom panel) mice.

J. Representative confocal images of fibrin (green), CD31 (red) and DAPI (blue) immunostaining in the brain vasculature of WT (top panel) and Glut1iECKO (bottom panel) mice

K. Quantification of the number of cadaverine 555 positive brain parenchyma cells in WT (blue) and Glut1iECKO (orange) mice. (n=13 from 3 independent experiments. Error bars indicate the standard error of the mean (SEM) from unpaired Student's t test.)

L. Quantification of extravascular fibrin deposits in WT (blue) and Glut1iECKO (orange) mice. (n=9 from 3 independent experiments. Error bars indicate the standard error of the mean (SEM) from unpaired Student's t test.)

M. Quantification of lactate in the cerebrospinal fluid of WT (blue) and Glut1iECKO (orange) mice. (n=3 from 3 independent experiments. Error bars indicate the standard error of the mean (SEM) from unpaired Student's t test.)