Differential protein expression in rat trigeminal ganglia during inflammation.
A proteomics approach was used to investigate global protein changes in rat sensory ganglia exposed to pro-inflammatory stimuli. Inflammation was provoked in vivo by injecting selected facial areas with Freunds Complete Adjuvant (FCA), or by stimulating freshly isolated trigeminal ganglia ex vivo with pro-inflammatory mediators (interleukin-1 beta, tumor necrosis factor-alpha, interferon-gamma). Following two-dimensional gel electrophoresis, silver staining and mass spectrometry, a protein reference map and database was generated for rat trigeminal ganglia, which to our knowledge is the first to be reported. Sixty-seven out of 85 selected protein spots were successfully identified using matrix-assisted laser desorption/ionization mass spectrometry. This reference map was used to monitor changes in the ganglia proteome induced during inflammation in vivo and ex vivo. In vivo we found that FCA treatment specifically induced differential protein expression of two unidentified protein spots and, to a lower extent, of beta-tubulin. Image analysis of ganglia treated ex vivo with the cocktail of cytokines indicated that some of the changes in the protein population were also observed in vivo after FCA treatment. If the cytokine stimulation was performed in the presence of acetaminophen (paracetamol), the drug seemed to reverse the effects of cytokine treatment for at least some protein spots, restoring the same protein pattern observed in control samples.
Friso
G
G
Departments of Molecular Sciences and Bioscience, AstraZeneca RD Södertälje, Sweden. gf32@cornell.edu
Kaiser
L
L
Raud
J
J
Wikström
L
L
D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D066298:In Vitro Techniques; D018836:Inflammation Mediators / Q000494:pharmacology; D008297:Male; D013058:Mass Spectrometry; D009419:Nerve Tissue Proteins / Q000302:isolation & purification / Q000378:metabolism*; D009443:Neuritis / Q000139:chemically induced / Q000378:metabolism*; D010449:Peptide Mapping; D020543:Proteome; D051381:Rats; D011919:Rats, Inbred Strains; D012668:Trigeminal Ganglion / Q000187:drug effects / Q000378:metabolism*
D018836:Inflammation Mediators; D009419:Nerve Tissue Proteins; D020543:Proteome