Expression, binding, and signaling properties of CRF2(a) receptors endogenously expressed in human retinoblastoma Y79 cells: passage-dependent regulation of functional receptors.

Eric Gutknecht, J Van Der Linden, Georges Vauquelin, R L Hauger, F M Dautzenberg

Onderzoeksoutput: Articlepeer review

16 Citaten (Scopus)

Samenvatting

Endogenous expression of the corticotropin-releasing factor type 2a receptor (CRF2(a)) but not CRF2(b) and CRF2(c) was observed in higher passage cultures of Y79 retinoblastoma cells. Functional studies further demonstrated an increase in CRF2(a) mRNA and protein levels with higher passage numbers (bigger than 25 passages). Although the CRF1 receptor was expressed at higher levels than the CRF2(a) receptor, both receptors were easily distinguishable from one another by selective receptor ligands. CRF1-preferring or non-selective agonists such as CRF, urocortin 1 and sauvagine stimulated cyclic AMP (cAMP) production in Y79 to maximal responses of 110 pmoles/ 105 cells, whereas the exclusive CRF2 receptor-selective agonists urocortin 2 and urocortin 3 stimulated cAMP production to maximal responses of 25-30 pmoles/105 cells. Urocortin 2 and urocortin 3-mediated cAMP stimulation was potently blocked by the 300-fold selective CRF2 antagonist antisauvagine (IC50 = 6.5plusmin1.6 nM), whereas the CRF1-selective antagonist NBI 27914 only blocked cAMP responses at concentrations >10 muM. When the CRF1-preferring agonist ovine CRF was used to activate cAMP signaling, NBI 27914 (IC50 = 38.4plusmin3.6 nM) was a more potent inhibitor than antisauvagine (IC50 = 2.04plusmin0.2 muM). Finally, urocortin 2 and 3 treatment potently and rapidly desensitized the CRF2 receptor responses in Y79 cells. These data demonstrate that Y79 cells express functional CRF1 and CRF2(a) receptors, and that the CRF2(a) receptor protein is upregulated during prolonged culture.
Originele taal-2English
Pagina's (van-tot)926-936
Aantal pagina's11
TijdschriftJournal of Neurochemistry
Volume104
StatusPublished - 2008

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