Proteomic analysis of phosphotyrosyl proteins in morphine-dependent rat brains

Seong Youl Kim, Nuannoi Chudapongse, Sang Lee, Michael C. Levin, Jae Taek Oh, Hae Joon Park, Ing K. Ho

Research output: Contribution to journalArticle

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Abstract

Morphine has been used as a potent analgesic, having a high propensity to induce tolerance and physical dependence following their repeated administration. Although the mechanisms that underlie the development of dependence on morphine remain unclear, previous studies suggested that phosphorylations of diverse types of cellular proteins are crucial determinants of the neuroadaptive mechanisms associated with morphine dependence. Thus, understanding global phosphorylation events induced by chronic morphine administration is essential for understanding the complex signaling mechanisms of morphine dependence. This study characterized the alteration of tyrosine phosphorylation of frontal cortical proteins in morphine-dependent rat brains using a proteomic approach. Dependence was produced by continuous intracerebroventricular (i.c.v.) infusion of morphine (26 nmol/μl/h) for 72 h via osmotic minipumps in rats. Phosphotyrosyl (p-Tyr) protein spots in brain frontal cortical regions were detected by two-dimensional electrophoresis (2-DE) and immunoblotting with anti-p-Tyr-specific antibodies. The protein spots showing significant changes in tyrosine phosphorylation were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Similar patterns of protein expression were detected by 2-DE gels in morphine-dependent and saline-treated control rat brains. However, phosphotyrosine 2-DE images of the frontal cortical proteins from saline-treated control and morphine-dependent rat brains were apparently different. The densities of most matched p-Tyr protein spots were increased in morphine-dependent rat brains compared with that of control samples. Additional p-Tyr protein spots were detected in 2-DE image of morphine-dependent rat brains. Fifty of p-Tyr protein spots, corresponding to 40 different proteins, were identified from 2-DE gels of morphine-dependent rat brains. The identified proteins include enzymes, cytoskeletal proteins, cell signaling molecules, and other proteins. In conclusion, the first available phosphotyrosine proteomic resources of morphine dependence were established using an animal model. The findings illustrate the potential of proteomics as an effective technique for studying phosphorylation events of morphine dependence in brains.

Original languageEnglish (US)
Pages (from-to)58-70
Number of pages13
JournalMolecular Brain Research
Volume133
Issue number1
DOIs
StatePublished - Jan 5 2005

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Proteomics
Morphine
Morphine Dependence
Brain
Proteins
Phosphorylation
Phosphotyrosine
Tyrosine
Gels
Intraventricular Infusions
Cytoskeletal Proteins
Immunoblotting
Analgesics
Electrophoresis
Mass Spectrometry
Lasers
Animal Models
Antibodies

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cellular and Molecular Neuroscience

Cite this

Proteomic analysis of phosphotyrosyl proteins in morphine-dependent rat brains. / Kim, Seong Youl; Chudapongse, Nuannoi; Lee, Sang; Levin, Michael C.; Oh, Jae Taek; Park, Hae Joon; Ho, Ing K.

In: Molecular Brain Research, Vol. 133, No. 1, 05.01.2005, p. 58-70.

Research output: Contribution to journalArticle

Kim, Seong Youl ; Chudapongse, Nuannoi ; Lee, Sang ; Levin, Michael C. ; Oh, Jae Taek ; Park, Hae Joon ; Ho, Ing K. / Proteomic analysis of phosphotyrosyl proteins in morphine-dependent rat brains. In: Molecular Brain Research. 2005 ; Vol. 133, No. 1. pp. 58-70.
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