Cholecystokinin-GABA interactions in rodent cortex: analyses of cholecystokinin effects on K(+)- and L-glutamate-induced release of [3H]GABA from rat cortical slices and cultured mouse cortical neurones.
Neurones of the cerebral cortex immunoreactive for the neuropeptide, cholecystokinin (CCK), also invariably contain GABA. Hence CCK is believed to modulate some aspect of GABAergic synaptic activity. The present study therefore investigated the effects of CCK on basal, K(+)- and L-glutamate-induced release of [3H]GABA from slices of rat neocortex and cultured murine neocortical neurones. Rat neocortical prisms loaded with [3H]GABA (10 nM) were superfused with Krebs-Henseleit buffer and stimulated twice (S1 and S2, 2 min) with K+ (30 mM). Release associated with each stimulus was measured and expressed relative to basal release (R1 and R2). The effects of non-selective and CCKB selective agonists, CCK-8S and CCK-4, respectively, on basal and K(+)-induced release of [3H]GABA were subsequently assessed by alternately including the peptides in S2 and comparing R2/R1 and S2/S1 ratios to control experiments. Contrary to previous findings, CCK-8S (30 nM-1 microM) and CCK-4 (0.3 nM-1 microM) failed to influence basal or K(+)-induced release. In similar experiments, murine cortical neurones superfused with HEPES balanced salt buffer, released exogenous [3H]GABA upon stimulation (1 min) with either K+ (55 mM) or L-glutamate (30 microM). However, CCK-8S, CCK-4 (both 300 nM-1 microM) and the CCKB selective antagonist, L365,260 (1 microM), failed to influence basal, K(+)- or L-glutamate-induced release of [3H]GABA from these neurones when included in S2. These data therefore do not support the postulate that CCK acting via CCKA or CCKB receptors modulates release of GABA under the present experimental conditions. GABA-CCK interactions were not specifically studied because only L-glutamate (30 microM) significantly elevated release of CCK-like immunoreactivity (115% above basal) in murine cortical neurones: basal release of CCK was estimated to be 7 and 11 pM from neurones and slices, respectively. Further studies employing more rigorous stimulation and perhaps examining endogenous GABA release are necessary to fully investigate the co-release of CCK and GABA.
Hickling
Y M
YM
Department of Pharmacology, Monash University, Clayton, Victoria, Australia.
Cheung
N S
NS
Larm
J A
JA
Cowen
M S
MS
Shulkes
A
A
Beart
P M
PM
D000818:Animals; D001481:Basal Metabolism; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002766:Cholecystokinin / Q000378:metabolism / Q000494:pharmacology*; D018698:Glutamic Acid / Q000494:pharmacology*; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D011188:Potassium / Q000494:pharmacology*; D011869:Radioligand Assay; D051381:Rats; D017207:Rats, Sprague-Dawley; D011949:Receptors, Cholecystokinin / Q000187:drug effects; D012844:Sincalide / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D013758:Tetragastrin / Q000378:metabolism / Q000494:pharmacology; D014316:Tritium; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
C090019:8-sulfocholecystokinin octapeptide; D011949:Receptors, Cholecystokinin; D013758:Tetragastrin; D014316:Tritium; D018698:Glutamic Acid; D005680:gamma-Aminobutyric Acid; D002766:Cholecystokinin; D012844:Sincalide; D011188:Potassium