Acute ethanol-induced adenosine diphosphate ribosylation regulates the functional activity of rat striatal pertussis toxin-sensitive g proteins.
BACKGROUND
We demonstrated previously that striatal adenosine modulates ethanol-induced motor incoordination (EIMI) via adenosine A1 receptors coupled to pertussis toxin (PT)-sensitive G protein and adenylyl cyclase-cyclic adenosine monophosphate (cAMP). Additionally, intrastriatal (IST) PT antagonizes EIMI and its potentiation by the adenosine A1 agonist N-cyclohexyladenosine; it also inhibits cAMP concentration.


METHODS
Guide cannulas were stereotaxically implanted for IST pretreatment with PT followed 5 days later by IST of N-cyclohexyladenosine and intraperitoneal ethanol. The adenosine diphosphate (ADP) ribosylation reaction involved PT-catalyzed [P]nicotinamide adenine dinucleotide (NAD) labeling of rat striatal membranes. Antagonism of EIMI (Rotorod method) after IST microinfusion of PT was investigated to determine whether it was due to a decrease in the functional activity of G proteins due to ADP ribosylation of the Gialpha subunit caused it.


RESULTS
Striatal membranes from IST PT (0.5 microg)-treated animals exhibited significantly attenuated (up to 90%) in vitro ADP ribosylation with [P]NAD. Striatal membranes from animals injected with ethanol (1.5 g/kg intraperitoneally) exhibited statistically significant increase (11%) in in vitro ADP ribosylation. Similarly, ethanol (50 mM) added to striatal membranes from untreated animals produced significant stimulation of in vitro ADP ribosylation. The decrease in the functional activity of G proteins due to ADP ribosylation of the Gialpha subunit after IST PT was functionally correlated with marked attenuation in EIMI, as observed previously. This finding suggests a blockade of functional activity of PT-sensitive striatal Gi/Go proteins (i.e., fewer available sites for labeled NAD incorporation). The in vivo ethanol results indicate that it must have caused an increase in the ribosylation capacity of Gialpha in vivo (i.e., increased Gi activity). Increased ADP ribosylation by in vitro ethanol increases Gi/Go activity, consistent with EIMI, as previously reported.


CONCLUSIONS
The results provide biochemical evidence of an ethanol-induced increase in ADP ribosylation of Gialpha causing a decrease in the functional activity of G proteins coupled via Gi/Go to adenylyl cyclase-cAMP. These results confirm the previously observed antagonism of EIMI by PT (IST).
Dar
M Saeed
MS
Department of Pharmacology, Brody School of Medicine, East Carolina University, Greenville, North Carolina 27858, USA. darm@mail.ecu.edu
Meng
Z-H
ZH
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D000818:Animals; D003342:Corpus Striatum / Q000187:drug effects* / Q000378:metabolism; D000431:Ethanol / Q000008:administration & dosage*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D008297:Male; D037342:Pertussis Toxin / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D000246:Adenosine Diphosphate Ribose; D000431:Ethanol; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins