Extracellular metabolism of adenine nucleotides and adenosine in the innervated skeletal muscle of the frog.
The effects of coformycin, alpha,beta-methylene ADP, dipyridamole in the absence and presence of erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA), nitrobenzylthioinosine (NBTI), mioflazine and ouabain on the metabolic pathways of exogenously applied ATP and its metabolites in the frog innervated sartorius muscle were investigated. ATP catabolism yielded ADP, AMP, IMP, adenosine and inosine; the ecto-ATPase in situ was shown to be Ca(2+)- or Mg(2+)-activated with a Kmapp for ATP of 767 +/- 48 microM. AMP catabolism yielded IMP, adenosine and inosine; inosine was formed from either exogenous IMP or exogenous adenosine. Catabolism of AMP into IMP was blocked by coformycin, which enhanced adenosine and inosine formation from AMP. alpha,beta-Methylene ADP blocked adenosine formation from AMP and inosine formation from IMP; formation of IMP from AMP was enhanced by alpha,beta-methylene ADP. Complete blockade of AMP degradation was achieved with the simultaneous use of coformycin and alpha,beta-methylene ADP. Dipyridamole attenuated but did not completely block extracellular adenosine removal and inosine appearance in the bath. EHNA, applied in the presence of dipyridamole, did not cause any further attenuation of extracellular adenosine removal. Mioflazine, NBTI and ouabain did not affect adenosine disappearance from the bath. The results suggest that, in the frog innervated sartorius muscle, ATP can be sequentially catabolized into AMP which is then catabolized either into IMP or into adenosine. This extracellular degradation of AMP into IMP might then constitute a shunt-like mechanism to control the levels of adenosine formed from adenine nucleotides.
Cunha
R A
RA
Gulbenkian Institute of Science, Laboratory of Pharmacology, Oeiras Codex, Portugal.
Sebastião
A M
AM
D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000227:Adenine Nucleotides / Q000378:metabolism*; D000241:Adenosine / Q000378:metabolism*; D000243:Adenosine Deaminase / Q000378:metabolism; D058892:Adenosine Deaminase Inhibitors; D000244:Adenosine Diphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002412:Cations / Q000494:pharmacology; D003070:Coformycin / Q000494:pharmacology; D004176:Dipyridamole / Q000494:pharmacology; D066298:In Vitro Techniques; D007291:Inosine Monophosphate / Q000378:metabolism; D007700:Kinetics; D009132:Muscles / Q000294:innervation / Q000378:metabolism*; D010879:Piperazines / Q000494:pharmacology; D011895:Rana ridibunda; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors; D013868:Thioinosine / Q000031:analogs & derivatives / Q000494:pharmacology
D000227:Adenine Nucleotides; D058892:Adenosine Deaminase Inhibitors; D002412:Cations; D010879:Piperazines; C523965:alpha,beta-methyleneadenosine 5'-diphosphate; D003070:Coformycin; D007291:Inosine Monophosphate; D013868:Thioinosine; C010192:9-(2-hydroxy-3-nonyl)adenine; D000244:Adenosine Diphosphate; D004176:Dipyridamole; D000255:Adenosine Triphosphate; C041518:mioflazine; D000243:Adenosine Deaminase; D000254:Sodium-Potassium-Exchanging ATPase; C001789:4-nitrobenzylthioinosine; D000225:Adenine; D000241:Adenosine