Proteomic analysis of phosphotyrosyl proteins in the rat brain: effect of butorphanol dependence.
Butorphanol (17-cyclobutylmethyl-3,14-dihydroxymorphinan) tartrate (Stadol) is a mixed agonist-antagonist opioid analgesic agent that is about five to seven times as potent as morphine in analgesic effects. The chronic use of butorphanol produces physical dependence in humans and animals. Phosphorylation plays a very important role in developing butorphanol dependence; however, global phosphorylation events induced by chronic butorphanol administration have not been reported. The aim of this study is to determine the alteration of tyrosine phosphorylation of brain frontal cortical proteins in butorphanol-dependent rats using a proteomic approach. Dependence was produced by continuous intracerebroventricular (i.c.v.) infusion of butorphanol (26 nmol/microl/hr) for 72 hr via osmotic minipump in rats. Similar patterns of protein expression were detected by two-dimensional electrophoresis (2-DE) in brain frontal cortex of butorphanol-dependent and saline-treated control rats. All 65 phosphotyrosyl (p-Tyr) protein spots detected in pH 3-10 phosphotyrosine 2-DE of control rat brains were detected in butorphanol-dependent rat brains. The densities of most p-Tyr protein spots were increased in butorphanol-dependent rat brains compared to saline-treated control samples. Eighteen additional p-Tyr protein spots were detected in pH 3-10 2-DE images of butorphanol-dependent rat brains. Immobilized pH strips with three different narrow pH ranges were examined to improve the resolution of p-Tyr proteins in 2-DE gels. Fifty-three p-Tyr protein spots were identified as known proteins involved in cell cytoskeleton, cell metabolism, and cell signaling. This proteomic approach can provide useful information for understanding the complex mechanism of butorphanol dependence in vivo.
Kim
Seong-Youl
SY
Department of Pharmacology and Toxicology, University of Mississippi Medical Center, Jackson, Mississippi 39216, USA.
Chudapongse
Nuannoi
N
Lee
Sang-Min
SM
Levin
Michael C
MC
Oh
Jae-Taek
JT
Park
Hae-Joon
HJ
Ho
Ing K
IK
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002077:Butorphanol / Q000008:administration & dosage*; D008297:Male; D009293:Opioid-Related Disorders / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D011506:Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley
D011506:Proteins; D019000:Phosphotyrosine; D002077:Butorphanol