Functional corticotropin-releasing factor receptors in human neuroblastoma cells.
The present study examined the presence of functional corticotropin-releasing factor (CRF) receptors in IMR-32 neuroblastoma cells. [125I]Tyro-ovine CRF binding was linear with increasing protein concentrations, saturable, reversible and of high affinity. Scatchard analysis indicated a Kd of approximately 0.8 nM and a Bmax of approximately 32 fmol/mg protein. Competition studies with CRF and related peptides revealed a pharmacological profile characteristic of the CRF1 receptor subtype. CRF stimulated cAMP production in a dose-dependent manner with an apparent EC50 of approximately 4 nM. In addition, the putative CRF receptor antagonist alpha-helical CRF9-41 dose-dependently inhibited CRF stimulated (10 nM) cAMP production with an IC50 of approximately 60 nM. CRF treatment down regulated its own receptor while treatment with the protein kinase C activator, phorbol 12-myristate 13-acetate (PMA), increased CRF binding in neuroblastoma cells. Taken together, these data demonstrate the utility of the human neuroblastoma cell line for functional studies on CRF receptors and suggest that CRF may play a regulatory role in the pathophysiology of human neuroblastoma.
Dieterich
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Neurocrine Biosciences, Inc., San Diego, CA 92037, USA.
DeSouza
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EB
D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D001665:Binding Sites / Q000502:physiology; D001667:Binding, Competitive / Q000502:physiology; D002417:Cattle; D003346:Corticotropin-Releasing Hormone / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects; D006801:Humans; D007457:Iodine Radioisotopes; D008565:Membrane Proteins / Q000378:metabolism; D009447:Neuroblastoma; D010455:Peptides / Q000378:metabolism / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism; D051381:Rats; D018019:Receptors, Corticotropin-Releasing Hormone / Q000032:analysis / Q000378:metabolism*; D012680:Sensitivity and Specificity; D012756:Sheep; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000201:enzymology
D007457:Iodine Radioisotopes; D008565:Membrane Proteins; D010455:Peptides; D018019:Receptors, Corticotropin-Releasing Hormone; D003346:Corticotropin-Releasing Hormone; D000242:Cyclic AMP; D011493:Protein Kinase C; D000262:Adenylyl Cyclases