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Mu-, kappa3-, and ORL-1-receptor mediated activation of p42 and p44 mitogen-activated protein kinases in BE -C and SH-SY5Y human neuroblastoma cells

Mu-, kappa3-, and ORL-1-receptor mediated activation of p42 and p44 mitogen-activated protein kinases in BE -C and SH-SY5Y human neuroblastoma cells

Society for Neuroscience Abstracts 26(1-2): Abstract No -434 8

mu (DAMGO), kappa3 (naloxone benzoylhydrazone; NalBzoH) and ORL-1 (orphaninFQ/nociceptin; OFQ/N) receptors induce rapid, short-term and dose-dependent activation of p42/p44 mitogen-activated protein kinases (MAPK), in BE(2)-C and SH-SY5Y human neuroblastoma cell lines that endogenously express mu, delta, kappa3 and ORL-1 receptors and display the morphological, neurochemical and electrophysiological characteristics of sympathetic neurons. DAMGO- and NalBzoH-mediated increases in MAPK activity were blocked by CTAP (100 nM) and Mr2266 (100 nM), respectively. The dose-dependent activation of MAPK by DAMGO, but not by NalBzoH, in CHO-MOR cells confirmed the kappa3 receptor-mediated action of NalBzoH. All agonist-mediated increases in MAPK activity were completely abolished upon pretreatment with pertussis toxin (100 ng/ml) and were also blocked by the phosphotidylinositol 3-kinase inhibitor, wortmannin (10 muM). DAMGO- and OFQ/N-mediated MAPK activation was also attenuated by the protein kinase C inhibitor, chelerythrine (1 muM), while NalBzoH-mediated MAPK activition was blocked by the protein kinase A inhibitor, H-9 (2 muM). Chronic activation (1-3 days) of mu and kappa3, but not ORL-1 receptors, led to an upregulation of tyrosine hydroxylase (an enzyme known to be upregulated as an intracellular adaptation upon chronic morphine exposure). These responses were attenuated upon pretreatment with the MEK-1 inhibitor, PD98059 (50 muM), suggesting a role for MAPK in the development of tolerance and dependence upon chronic activation of these receptors.

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