Hypoxic response of synaptosomal proteins in term guinea pig fetuses.
Early events in the hypoxia-induced response trigger tyrosine phosphorylation cascades involving a large number of enzymes and substrates. The resolving power of advanced two-dimensional gel electrophoresis, followed by immunoblotting with specific antibodies to phosphotyrosine, has been used to analyze hypoxia-induced modifications in guinea pig brain synaptosomes. These procedures, in conjunction with computer-aided image analysis, are useful in the differential display of gene products, providing comparison at the level of posttranslationally modified products. Studies were performed in cerebral cortical synaptosomes from three normoxic and three hypoxic newborn guinea pigs. To filter off background noise consisting of nonreproducible migrating protein spots, only reproducible features of electrophoretic patterns were considered. Immunoreactivity patterns obtained with anti-phosphotyrosine antibodies proved to be different in normoxic and hypoxic synaptosomes: of a total of 130 immunoreactive spots, 49 were tyrosine-phosphorylated in hypoxic synaptosomes only and 20 in the normoxic ones only. Our data suggest that hypoxia extensively remodels the signaling pathway by switching off tyrosine phosphorylation of some cellular components (i.e., alpha-internexin) and switching on tyrosine phosphorylation of some other proteins (i.e., heat shock cognate 70, aconitase, 2',3'-cyclic nucleotide 3'-phosphodiesterase, and pyruvate kinase).
Buonocore
G
G
Institute of Preventive Pediatrics and Neonatology, Italy.
Liberatori
S
S
Bini
L
L
Mishra
O P
OP
Delivoria-Papadopoulos
M
M
Pallini
V
V
Bracci
R
R
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000032:analysis; D000154:Aconitate Hydratase / Q000032:analysis; D000818:Animals; D002352:Carrier Proteins / Q000032:analysis; D002540:Cerebral Cortex / Q000196:embryology* / Q000648:ultrastructure; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D005311:Fetal Hypoxia / Q000378:metabolism*; D006168:Guinea Pigs; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis; D006801:Humans; D015151:Immunoblotting; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D019000:Phosphotyrosine / Q000032:analysis; D011247:Pregnancy; D011770:Pyruvate Kinase / Q000032:analysis; D015398:Signal Transduction; D016622:Silver Staining; D013574:Synaptosomes / Q000378:metabolism*
D002352:Carrier Proteins; D018840:HSP70 Heat-Shock Proteins; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C046643:alpha-internexin; D019000:Phosphotyrosine; D011770:Pyruvate Kinase; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases; D000154:Aconitate Hydratase