Evidence against adenosine analogues being agonists at the growth hormone secretagogue receptor.
Adenosine and adenosine analogues have been reported to act as agonists or partial agonists at the growth hormone secretagogue receptor 1a (GHSR1a). We have re-examined this question. A concentration-dependent increase in intracellular calcium concentration ([Ca(2+)](i)) was observed in GHSR1a transfected HEK 293-EBNA cells stimulated with adenosine (EC50: 0.2 microM) or 2-chloroadenosine (EC50: 1.1 microM) but also in untransfected HEK 293-EBNA cells stimulated with 2-chloroadenosine (EC50: 0.67 microM) or 5'-N-ethylcarboxamidoadenosine (NECA) (EC50: 0.045 microM). These findings support endogenous expression of adenosine receptors, presumably A(2B) receptors in HEK 293-EBNA cells. In GHSR1a transfected CHO cells, lacking adenosine receptors, the GHSR1a agonist hGhrelin (EC50: 2.4 nM) increased [Ca(2+)](i), but no effects of adenosine, 2-chloroadenosine or NECA were detected. An inverse agonist of GHSR1a, [d-Arg-1, d-Phe-5, d-Trp-7, 9, Leu-11] substance P, reduced hGhrelin effects but adenosine, 2-chloroadenosine or 1,3-dipropyl-8-cyclopentylxanthine (DPCPX) did not. NECA increased the [Ca(2+)](i) in co-transfected (GHSR1a and A(2B) receptor) CHO cells (EC50: 0.053 microM), but no additive or synergistic effects on [Ca(2+)](i) or cAMP formation were observed after stimulation with NECA in the absence or in the presence of hGhrelin. In binding studies on GHSR1a transfected CHO cell membranes, [(125)I]-hGhrelin binding could be displaced by the GHSR1a agonist MK-0677 (IC50: 0.34 nM), hGhrelin (IC50: 1.5 nM), and the substance P analogue (IC50: 0.64 microM) but not by adenosine or 2-chloroadenosine. We conclude that adenosine and analogues do not act as agonists or partial agonists at the GHSR1a and that cross-talk between the GHSR1a and A(2B) receptors is limited.
Department of Physiology and Pharmacology, Section of Molecular Pharmacology, Karolinska Institutet, S-171 77 Stockholm, Sweden. stina.johansson@fyfa.ki.se
Stina
S
Johansson
Bertil B
BB
Fredholm
Charlotta
C
Hjort
Torbjörn
T
Morein
Björn
B
Kull
Ping-Sheng
PS
Hu
D000241:Adenosine / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D016466:CHO Cells; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism; D054439:Ghrelin; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D036361:Peptide Hormones / Q000378:metabolism; D011869:Radioligand Assay; D043562:Receptors, G-Protein-Coupled / Q000819:agonists*; D054440:Receptors, Ghrelin; D015398:Signal Transduction
D054439:Ghrelin; D036361:Peptide Hormones; D043562:Receptors, G-Protein-Coupled; D054440:Receptors, Ghrelin; D000242:Cyclic AMP; D005978:Glutathione; D000241:Adenosine; D002118:Calcium