Opposing facilitatory and inhibitory modulation of glutamate release elicited by cAMP production in cerebrocortical nerve terminals (synaptosomes).
Activation of cAMP-protein kinase A (PKA) is widely reported to facilitate synaptic transmission. Here, we examined the presynaptic loci of PKA action using isolated nerve terminals (synaptosoms). The adenylyl cyclase (AC) activator, forskolin, failed to have any effect on 4-aminopyridine (4-AP)-evoked glutamate release, when added alone. However, in the presence of the alkylxanthine, IBMX, forskolin strongly facilitated glutamate release. This potentiation of release was blocked by the PKA inhibitors Rp-cAMPS and H7. Given that IBMX has dual activity, antagonizing adenosine receptors as well as inhibiting cAMP phosphodiesterase, we examined the effect of a selective adenosine A(1) receptor antagonist, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) and RO20-1724, a specific phosphodiesterase inhibitor. Both unmasked the forskolin-mediated modulation of glutamate release. Conversely, the adenosine analogue, N(6)-cyclohexyladenosine (CHA), reversed the facilitation induced by forskolin+RO20-1724. Adenosine A(1) receptor activation, therefore, appears to curtail cAMP/PKA-induced potentiation of glutamate release. Looking at the targets for cAMP/PKA-mediated potentiation of glutamate release, while synaptosomal excitability was only marginally increased, basal and 4-AP-evoked-increases in [Ca(2+)](c) were substantially enhanced by forskolin+IBMX. Moreover, glutamate release elicited by Ca(2+)-ionophore (ionomycin)-induced Ca(2+)-entry was facilitated by forskolin+IBMX. cAMP/PKA-mediated facilitation of glutamate release may therefore involve modulation of Ca(2+)-entry, as well as downstream events controlling synaptic vesicle recruitment and exocytosis.
Wang
S J
SJ
Department of Pharmacology, University College London, Medawar Building, Gower Street, UK.
Sihra
T S
TS
D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism*; D000242:Cyclic AMP / Q000096:biosynthesis*; D018698:Glutamic Acid / Q000378:metabolism*; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D009433:Neural Inhibition / Q000502:physiology; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013574:Synaptosomes / Q000187:drug effects / Q000378:metabolism*
D018698:Glutamic Acid; D000242:Cyclic AMP