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Glucocorticoids plus opioids up-regulate genes that influence neuronal function

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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 μ-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 (β1-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 μ-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.

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Acknowledgments

The authors thank Jenelle Jones, M.S. for assisting in the culturing of the SH-SY5Y human neuroblastoma cells and Western immunoblotting analysis. This work was supported by National Institute on Drug Abuse grants U24-DA12385 and R01-DA13770 (K.E.V.), K05-DA00182 (A.C.H.) and the National Center on Minority Health and Health Disparities P21-MD00175.

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  1. Gregg R. Ward

    Present address: Department of Biology, Division of Natural Sciences & Mathematics, Saint Augustine’s College, Raleigh, NC, 27610, USA

  2. Krystal T. Dempsey

    Present address: Biogen Idec, Research Triangle Park, NC, 27709, USA

Authors and Affiliations

  1. Neuroscience of Drug Abuse Research Program, Julius L. Chambers Biomedical/Biotechnology Research Institute, North Carolina Central University, Durham, NC, 27707, USA

    Gregg R. Ward, Steven O. Franklin, Tonya M. Gerald, Krystal T. Dempsey, Darrel E. Clodfelter Jr. & Allyn C. Howlett

  2. Department of Pharmacology, Pennsylvania State University College of Medicine/Milton S. Hershey Medical Center, Hershey, PA, 17033, USA

    Dan J. Krissinger, Kruti M. Patel & Kent E. Vrana

  3. Department of Physiology & Pharmacology, Wake Forest University Health Sciences, Medical Center Blvd., Winston-Salem, NC, 27157, USA

    Allyn C. Howlett

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  1. Gregg R. Ward
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Ward, G.R., Franklin, S.O., Gerald, T.M. et al. Glucocorticoids plus opioids up-regulate genes that influence neuronal function. Cell Mol Neurobiol 27, 651–660 (2007). https://doi.org/10.1007/s10571-007-9151-3

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