Protein profiling the effects of in vitro hyperoxic exposure on fetal rabbit lung.
The aim of this study was to test the hypothesis that acute in vitro exposure of prematurely delivered fetal rabbit lungs to hyperoxic conditions will induce the expression of an adaptive cassette of proteins that mediates antioxidant and inflammatory processes. To test this hypothesis, ex situ fetal rabbit lung explants were prepared from New Zealand white rabbits delivered by cesarean section on day 29 of gestation and incubated under air (21% O2; 5% CO2) or hyperoxic (95% O2; 5% CO2) atmospheres. Total tissue protein was extracted following incubation and subjected to 2-DE. Using this technique, 1500-2000 protein spots were resolved per gel. Treatment-dependent, differentially expressed proteins were identified by image analysis (Melanie II) and MALDI-TOF MS and MALDI-MS/MS. The analysis identified 12 protein spots that were differentially expressed by 1.5-fold or more (p<0.05) by exposure to hyperoxic conditions. Six of these differentially expressed proteins were identified as vimentin, annexin I, inorganic pyrophosphatase, prohibitin, an N-terminal fragment of ATP synthase and heat shock protein 27. The data obtained are consistent with the roles of these proteins in mediating cellular response to oxidative stress and in regulating cell proliferation.
Henschke
Phillip
P
Department of Neonatology, The Royal Women's Hospital, Melbourne, Australia.
Vorum
Henrik
H
Honoré
Bent
B
Rice
Gregory Edward
GE
D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D066298:In Vitro Techniques; D008168:Lung / Q000196:embryology / Q000378:metabolism*; D013058:Mass Spectrometry; D018384:Oxidative Stress; D010100:Oxygen / Q000502:physiology*; D010449:Peptide Mapping; D040081:Protein Array Analysis / Q000379:methods*; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D011817:Rabbits; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D011506:Proteins; D010100:Oxygen