TY - JOUR
T1 - Characterization of the Drosophila adenosine receptor
T2 - The effect of adenosine analogs on cAMP signaling in Drosophila cells and their utility for in vivo experiments
AU - Kucerova, Lucie
AU - Broz, Vaclav
AU - Fleischmannova, Jana
AU - Santruckova, Eva
AU - Sidorov, Roman
AU - Dolezal, Vladimir
AU - Zurovec, Michal
PY - 2012/5
Y1 - 2012/5
N2 - Adenosine receptors (AR) belonging to the G protein-coupled receptor family influence a wide range of physiological processes. Recent elucidation of the structure of human A2AR revealed the conserved amino acids necessary for contact with the Ado moiety. However, the selectivity of Ado analogs for AR subtypes is still not well understood. We have shown previously that the Drosophila adenosine receptor (DmAdoR) evokes an increase in cAMP and calcium concentration in heterologous cells. In this study, we have characterized the second-messenger stimulation by endogenous DmAdoR in a Drosophila neuroblast cell line and examined a number of Ado analogs for their ability to interact with DmAdoR. We show that Ado can stimulate cAMP but not calcium levels in Drosophila cells. We found one full and four partial DmAdoR agonists, as well as four antagonists. The employment of the full agonist, 2-chloroadenosine, in flies mimicked in vivo the phenotype of DmAdoR over-expression, whereas the antagonist, SCH58261, rescued the flies from the lethality caused by DmAdoR over-expression. Differences in pharmacological effect of the tested analogs between DmAdoR and human A2AR can be partially explained by the dissimilarity of specific key amino acid residues disclosed by the alignment of these receptors.
AB - Adenosine receptors (AR) belonging to the G protein-coupled receptor family influence a wide range of physiological processes. Recent elucidation of the structure of human A2AR revealed the conserved amino acids necessary for contact with the Ado moiety. However, the selectivity of Ado analogs for AR subtypes is still not well understood. We have shown previously that the Drosophila adenosine receptor (DmAdoR) evokes an increase in cAMP and calcium concentration in heterologous cells. In this study, we have characterized the second-messenger stimulation by endogenous DmAdoR in a Drosophila neuroblast cell line and examined a number of Ado analogs for their ability to interact with DmAdoR. We show that Ado can stimulate cAMP but not calcium levels in Drosophila cells. We found one full and four partial DmAdoR agonists, as well as four antagonists. The employment of the full agonist, 2-chloroadenosine, in flies mimicked in vivo the phenotype of DmAdoR over-expression, whereas the antagonist, SCH58261, rescued the flies from the lethality caused by DmAdoR over-expression. Differences in pharmacological effect of the tested analogs between DmAdoR and human A2AR can be partially explained by the dissimilarity of specific key amino acid residues disclosed by the alignment of these receptors.
KW - AdoR
KW - CG9753
KW - GPCR
KW - GloSensor
KW - calcium
KW - cyclic AMP
UR - http://www.scopus.com/inward/record.url?scp=84859625334&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84859625334&partnerID=8YFLogxK
U2 - 10.1111/j.1471-4159.2012.07701.x
DO - 10.1111/j.1471-4159.2012.07701.x
M3 - Article
C2 - 22353178
AN - SCOPUS:84859625334
SN - 0022-3042
VL - 121
SP - 383
EP - 395
JO - Journal of Neurochemistry
JF - Journal of Neurochemistry
IS - 3
ER -