title: Glucocorticoids plus opioids up-regulate genes that influence neuronal function.
abstract: (1) This study investigated the functional genomics of glucocorticoid and opioid receptor stimulation in cellular adaptations using a cultured neuronal cell model. (2) Human SH-SY5Y neuroblastoma cells grown in hormone-depleted serum were treated for 2-days with the glucocorticoid receptor-II agonist dexamethasone (30 nM); the mu-opioid receptor agonist [D-Ala2, N-Me-Phe4, Gly5-ol]-Enkephalin acetate (DAMGO; 1 nM); or dexamethasone (30 nM) plus DAMGO (1 nM). RNA was extracted; purified, reverse transcribed, and labeled cDNA was hybridized to a 10,000-oliogonucleotide-array human gene chip. Gene expression changes that were significantly different between treatment groups and were of interest due to biological function were verified by real-time reverse transcription polymerase chain reaction (RT-PCR). Five relevant genes were identified for which the combination of dexamethasone plus DAMGO, but neither one alone, significantly up-regulated gene expression (ANOVA, P < 0.05). (3) Proteins coded by the identified genes: FRS2 (fibroblast growth factor receptor substrate-2; CTNNB1 (beta1-catenin); PRCP (prolyl-carboxypeptidase); MPHOSPH9 (M-phase phosphoprotein 9); and ZFP95 (zinc finger protein 95) serve important neuronal functions in signal transduction, synapse formation, neuronal growth and development, or transcription regulation. Neither opioid, glucocorticoid nor combined treatments significantly altered the cell growth rate determined by cell counts and protein. (4) We conclude that sustained mu-opioid receptor stimulation accompanied by glucocorticoids can synergistically regulate genes that influence neuronal function. Future studies are warranted to determine if combined influences of glucocorticoid fluctuations and opioid receptor stimulation in vivo can orchestrate exagerated neuroadaptation to reinforcing drugs under chronic mild stress conditions.
authors.affiliation: Neuroscience of Drug Abuse Research Program, Julius L. Chambers Biomedical/Biotechnology Research Institute, North Carolina Central University, Durham, NC 27707, USA.
authors.forename: Gregg R
authors.initials: GR
authors.lastname: Ward
authors.forename: Steven O
authors.initials: SO
authors.lastname: Franklin
authors.forename: Tonya M
authors.initials: TM
authors.lastname: Gerald
authors.forename: Krystal T
authors.initials: KT
authors.lastname: Dempsey
authors.forename: Darrel E
authors.initials: DE
authors.lastname: Clodfelter
authors.forename: Dan J
authors.initials: DJ
authors.lastname: Krissinger
authors.forename: Kruti M
authors.initials: KM
authors.lastname: Patel
authors.forename: Kent E
authors.initials: KE
authors.lastname: Vrana
authors.forename: Allyn C
authors.initials: AC
authors.lastname: Howlett
mesh_terms: D000701:Analgesics, Opioid / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor; D003907:Dexamethasone / Q000494:pharmacology*; D004357:Drug Synergism; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)- / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D023281:Genomics; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D009447:Neuroblastoma; D009473:Neuronal Plasticity / Q000187:drug effects; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis; D017450:Receptors, Opioid, mu / Q000819:agonists; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D000701:Analgesics, Opioid; D005938:Glucocorticoids; D017450:Receptors, Opioid, mu; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; D003907:Dexamethasone