omega-agatoxin-iva and enadoline

The modulation of depolarization (4-aminopyridine, 2 mM)-evoked endogenous glutamate release by kappa-opioid receptor activation and blockade of voltage-dependent Ca2+ -channels has been investigated in synaptosomes prepared from rat and marmoset striatum. 4-Aminopyridine (4-AP)-stimulated, Ca2+ -dependent glutamate release was inhibited by enadoline, a selective kappa-opioid receptor agonist, in a concentration-dependent and norbinaltorphimine (nor-BNI, selective kappa-opioid receptor antagonist)-sensitive manner in rat (IC50 = 4.4+/-0.4 microM) and marmoset (IC50 = 2.9+/-0.7 microM) striatal synaptosomes. However, in the marmoset, there was a significant (approximately 23%) nor-BNI-insensitive component. In rat striatal synaptosomes, the Ca2+ -channel antagonists omega-agatoxin-IVA (P/Q-type blocker), omega-conotoxin-MVIIC (N/P/Q-type blocker) and omega-conotoxin-GVIA (N-type blocker) reduced 4-AP-stimulated, Ca2+ -dependent glutamate release in a concentration-dependent manner with IC50 values of 6.5+/-0.9 nM, 75.5+5.9 nM and 106.5+/-8.7 nM, respectively. In marmoset striatal synaptosomes, 4-AP-stimulated, Ca2+ -dependent glutamate release was significantly inhibited by omega-agatoxin-IVA (30 nM, 57.6+/-2.3%, inhibition), omega-conotoxin-MVIIC (300 nM, 57.8+/-3.1%) and omega-conotoxin-GVIA (1 microM, 56.7+/-2%). Studies utilizing combinations of Ca2+ -channel antagonists suggests that in the rat striatum, two relatively distinct pools of glutamate, released by activation of either P or Q-type Ca2+ -channels, exist. In contrast, in the primate there is much overlap between the glutamate released by P and Q-type Ca2+ -channel activation. Studies using combinations of enadoline and the Ca2+ -channel antagonists suggest that enadoline-induced inhibition of glutamate release occurs primarily via reduction of Ca2+ -influx through P-type Ca2+ -channels in the rat but via N-type Ca2+ -channels in the marmoset. In conclusion, the results presented suggest that there are species differences in the control of glutamate release by kappa-opioid receptors and Ca2+ -channels.

Topics: 4-Aminopyridine; Animals; Benzofurans; Calcium Channel Blockers; Calcium Channels; Callithrix; Glutamic Acid; In Vitro Techniques; Male; Naltrexone; Narcotic Antagonists; Neostriatum; omega-Agatoxin IVA; omega-Conotoxin GVIA; Peptides; Pyrrolidines; Rats; Rats, Sprague-Dawley; Receptors, Opioid, kappa; Spider Venoms; Synaptosomes; Tetrodotoxin