Mfsd2a (Major Facilitator Superfamily Domain Containing 2a) Attenuates Intracerebral Hemorrhage-Induced Blood-Brain Barrier Disruption by Inhibiting Vesicular Transcytosis

J Am Heart Assoc. 2017 Jul 19;6(7):e005811. doi: 10.1161/JAHA.117.005811.

Abstract

Background: Blood-brain barrier (BBB) disruption aggravates brain injury induced by intracerebral hemorrhage (ICH); however, the mechanisms of BBB damage caused by ICH remain elusive. Mfsd2a (major facilitator superfamily domain containing 2a) has been known to play an essential role in BBB formation and function. In this study, we investigated the role and underlying mechanisms of Mfsd2a in BBB permeability regulation after ICH.

Methods and results: Using ICH models, we found that Mfsd2a protein expression in perihematomal brain tissues was significantly decreased after ICH. Knockdown and knockout of Mfsd2a in mice markedly increased BBB permeability, neurological deficit score, and brain water contents after ICH, and these were rescued by overexpressing Mfsd2a in perihematomas. Moreover, we found that Mfsd2a regulation of BBB permeability after ICH correlated with changes in vesicle number. Expression profiling of tight junction proteins showed no differences in Mfsd2a knockdown, Mfsd2a knockout, and Mfsd2a overexpression mice. However, using electron microscopy following ICH, we observed a significant increase in pinocytotic vesicle number in Mfsd2a knockout mice and decreased the number of pinocytotic vesicles in mouse brains with Mfsd2a overexpression. Finally, using multiple reaction monitoring, we screened out 3 vesicle trafficking-related proteins (Srgap2, Stx7, and Sec22b) from 31 vesicle trafficking-related proteins that were markedly upregulated in Mfsd2a knockout mice compared with controls after ICH.

Conclusions: In summary, our results suggest that Mfsd2a may protect against BBB injury by inhibiting vesicular transcytosis following ICH.

Keywords: Mfsd2a; blood–brain barrier; intracerebral hemorrhage; vesicular transcytosis.

MeSH terms

  • Animals
  • Behavior, Animal
  • Blood-Brain Barrier / metabolism*
  • Blood-Brain Barrier / ultrastructure
  • Capillary Permeability*
  • Carrier Proteins / metabolism
  • Cerebral Hemorrhage / genetics
  • Cerebral Hemorrhage / metabolism*
  • Cerebral Hemorrhage / pathology
  • Cerebral Hemorrhage / prevention & control
  • Disease Models, Animal
  • Endothelial Cells / metabolism*
  • Endothelial Cells / ultrastructure
  • GTPase-Activating Proteins
  • Genetic Predisposition to Disease
  • Male
  • Membrane Transport Proteins / deficiency
  • Membrane Transport Proteins / genetics
  • Membrane Transport Proteins / metabolism*
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Phenotype
  • Qa-SNARE Proteins / metabolism
  • R-SNARE Proteins / metabolism
  • RNA Interference
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Signal Transduction
  • Symporters
  • Tight Junctions / metabolism
  • Tight Junctions / ultrastructure
  • Time Factors
  • Transcytosis*
  • Transport Vesicles / metabolism*
  • Transport Vesicles / ultrastructure

Substances

  • Carrier Proteins
  • GTPase-Activating Proteins
  • Membrane Transport Proteins
  • Mfsd2a protein, mouse
  • Qa-SNARE Proteins
  • R-SNARE Proteins
  • RNA, Small Interfering
  • Sec22b protein, mouse
  • Srgap2 protein, mouse
  • Symporters