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Proteomic analysis of chronic morphine treated SH‐SY5Y human neuroblastoma cells (803.10)
Author(s) -
Randell Kesa,
Taka Equar,
Goodman Carl
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.803.10
Subject(s) - sh sy5y , morphine , opioid , pharmacology , chemistry , (+) naloxone , creb , proteomics , neuroblastoma , receptor , medicine , biology , biochemistry , cell culture , genetics , gene , transcription factor
The development of opioid tolerance is a barrier to its clinical use. Continuous exposure to opioids, like morphine, induces several molecular and cellular changes at the receptor and signal transduction pathway. Chronic opioid treatment of differentiated SH‐SY5Y human neuroblastoma cells has been shown to be an excellent in vitro model for studying cellular changes observed after opioid exposure. The present study was conducted to gain a better understanding of alterations in protein expression induced after opioid treatment, using mass spectrometry based proteomics, which in recent years has had a major impact on investigating the molecular and cellular mechanism of protein‐protein interactions. Cells were treated with morphine (10 μM), naloxone (10 μM), or 1 hr naloxone (10 μM) pre‐treatment followed by morphine (10 μM) for 24 hours, harvested, and separated by 2‐D gel electrophoresis. Analysis of these gels revealed alterations in protein expression induced by morphine treatment. Proteins with the most significant changes following drug treatment were identified using MALDI‐TOF. Calmodulin, alpha‐enolase, stathmin, nucleoside diphosphate kinase A, and nucleophosmin were identified. These results form the basis for the analysis of differential protein expression in SH‐SY5Y cells. The proteins detected will further be investigated to determine their role in morphine tolerance. Grant Funding Source : NIH‐NIMHD G12MD007582