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Functional, endogenously expressed corticotropin‐releasing factor receptor type 1 (CRF 1 ) and CRF 1 receptor mRNA expression in human neuroblastoma SH‐SY5Y cells
Author(s) -
Schoeffter Philippe,
Feuerbach Dominik,
Bobimac Ionel,
Gazi Lucien,
Longato Rémy
Publication year - 1999
Publication title -
fundamental and clinical pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.655
H-Index - 73
eISSN - 1472-8206
pISSN - 0767-3981
DOI - 10.1111/j.1472-8206.1999.tb00007.x
Subject(s) - urocortin , receptor , medicine , endocrinology , sh sy5y , biology , corticotropin releasing hormone , peptide , messenger rna , neuroblastoma , cell culture , biochemistry , gene , genetics
— Corticotropin‐releasing factor (CRF) is a hypothalamic 41‐amino acid peptide which stimulates corticotropin (ACTH) release from the anterior pituitary and is also involved in the body response to stress. CRF, receptors represent a potential target for novel antidepressant/anxiolytic drugs. The aim of the present study was to search for a human cell line expressing native, functional CRF, receptors as a starting material for screening purposes. We identified CRF, receptors functionally coupled to cAMP formation in human neuroblastoma SH‐SY5Y cells. CRF induced concentration‐dependent increases in cAMP accumulation in SH‐SY5Y cells (maximal increase 6.9 ± 0.9 fold over basal values, n = 14). This effect was mimicked by related peptides with similar potencies: (mean pEC 50 value) human/rat CRF (8.63), rat urocortin (9.32), sauvagine (8.97), urotensin I (8.93), ovine CRF (8.81). The efficacies of these agonists were nearly the same, with the exception of ovine CRF which was slightly less efficacious (75% the E max of CRF). The responses to CRF were competitively antagonised by the following peptide fragments (mean pK B value): α‐helical‐CRF (9–41) (7.54), [D‐Phe 12 , Nle 21,38 , CαMeLeu 37 ]CRF (12–41) (8.36) and [D‐Tyr 12 ]astressin (9.49) and by the selective, non‐peptidic CRF, receptor antagonists, CP‐154,526 (7.76) and antalarmin (9.19). Estimation of receptor density by [ 125 I]Tyr 0 ‐ovine CRF saturation binding yielded a modest number of binding sites (B max 12 fmol/mg protein, K D 0.2 nM). Analysis of mRNA by reverse transcription‐polymerase chain reaction clearly revealed the presence of mRNA for CRF 1 receptors in SH‐SY5Y cells. A slight signal for CRF 2 receptor mRNA was also observed. We conclude that neuroblastoma SH‐SY5Y cells are endowed with native CRF 1 receptors positively coupled to cAMP formation. They therefore constitute a useful functional model for the search of CRF 1 selective compounds with potential anxiolytic/antidepressant activity.