Proteomic analysis of phosphotyrosyl proteins in the rat brain: Effect of butorphanol dependence

Abstract

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.

Genes and Pathways Co-associated with the Exposure to Multiple Drugs of Abuse, Including Alcohol, Amphetamine/Methamphetamine, Cocaine, Marijuana, Morphine, and/or Nicotine: a Review of Proteomics Analyses

“…Most of the proteins identified from proteomics analyses fall into several major functional categories, such as protein modification, energy metabolism, cell cytoskeleton, signal transduction, and vesicular trafficking; many of these proteins are involved in biological processes such as synaptic plasticity, learning, and memory. Similar patterns are also observed in analyses focused on phosphotyrosyl proteins in the frontal cortex of morphine-or butorphanoldependent rats [51,64].…”

Effects of chronic tramadol exposure on the zebrafish brain: A proteomic study

“…Energy metabolism Aldolase C Nicotine [52,53]; Alcohol [23,56]; Amphetamine [57]; Butorphanol [24]; Cocaine [17]; Heroin [21]; Morphine [11,58,62]; ATP synthase beta-chain Nicotine [52]; Amphetamine [57]; Alcohol [23,55]; Cocaine [59]; Heroin [21]; Morphine [11,13,14,58]; Creatine kinase B Alcohol [23,[54][55][56]60]; Methamphetamine [61]; Cocaine [17];…”

“…Signal transduction 14-3-3 proteins Nicotine [52]; Butorphanol [24]; Alcohol [23,55]; Cocaine [17,59]; Morphine [13,24,58]; Heroin [16];…”

“…Recently, 2D-PAGE, in combination with mass spectrometry has been widely used in the identification of specific biochemical markers for diagnostic, preventive and therapeutic intervention in drug addiction, including morphine [11][12][13][14][15], cocaine [16][17][18], nicotine [19,20], heroin [21], alcohol [22,23] and butorphanol [24]. The neurotoxic effects of drug abuse are often associated with oxidative stress, mitochondrial dysfunction, apoptosis and inhibition of neurogenesis, among other mechanisms [25].…”

Neuroproteomics of the Synapse and Drug Addiction

“…The effects of morphine and butorphanol (a mixed opioid agonist-antagonist) on phosphotyrosyl proteins in the rat frontal cortex have been analyzed by 2-DE followed by MALDI-TOF MS (Kim et al, 2004b(Kim et al, , 2005. Rats were given intracerebroventricular infusions of morphine or butorphanol for 72 h and sacrificed 6 h later.…”