Differential protein expression in brain capillary endothelial cells induced by hypoxia and posthypoxic reoxygenation.
Cerebral ischemia causes functional alteration of the blood-brain barrier, formed by brain capillary endothelial cells (BCEC). Changes in protein expression and activity of selected differentially expressed enzymes were investigated in BCEC subjected to hypoxia (24 h) alone or followed by a 24-h reoxygenation. BCEC proteins were isolated, separated by 2-DE, and identified by MALDI-MS. Computer-based 2-D gel analysis identified 21 up-regulated proteins and 4 down-regulated proteins after hypoxia alone and 9 proteins that were further up-regulated after posthypoxic reoxygenation. The expression of the majority of hypoxia-induced proteins was reduced toward control levels during reoxygenation. The most prominent changes were identified for glycolytic enzymes (e.g., phosphoglycerate kinase), proteins of the ER (e.g., calreticulin), and cytoskeletal (e.g., vimentin) proteins. The results indicate that BCEC respond to hypoxia/reoxygenation by adaptive up-regulation of proteins involved in the glycolysis, protein synthesis, and stress response.
Haseloff
Reiner F
RF
Institute of Molecular Pharmacology, Berlin, Germany. haseloff@fmp-berlin.de
Krause
Eberhard
E
Bigl
Marina
M
Mikoteit
Kerstin
K
Stanimirovic
Danica
D
Blasig
Ingolf E
IE
D000818:Animals; D001921:Brain / Q000098:blood supply*; D002196:Capillaries / Q000166:cytology*; D015687:Cell Hypoxia / Q000502:physiology*; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000032:analysis; D013058:Mass Spectrometry; D008970:Molecular Weight; D010100:Oxygen / Q000502:physiology*; D010449:Peptide Mapping; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D011770:Pyruvate Kinase / Q000032:analysis; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000378:metabolism / Q000494:pharmacology; D015854:Up-Regulation
D011506:Proteins; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating); D011770:Pyruvate Kinase; D014357:Trypsin; D010100:Oxygen