title: Large conductance calcium-activated potassium channels: their expression and modulation of glutamate release from nerve terminals isolated from rat trigeminal caudal nucleus and cerebral cortex.
abstract: Large conductance, calcium-activated potassium channels [big potassium (BK) channel] consist of a tetramer of pore-forming α-subunit and distinct accessory β-subunits (β1-4) that modify the channel's properties. In this study, we analyzed the effects of BK channel activators and blockers on glutamate and γ-aminobutyric acid (GABA) release from synaptosomes isolated from the cerebral cortices or trigeminal caudal nuclei (TCN) of rats. Real-time polymerase chain reaction was used to characterize BK channel α and β(1-4) subunit expression in the cortex and in the trigeminal ganglia (TG), whose neurons project primary terminal afferents into the TCN. Immunocytochemistry was used to localize these subunits on cortical and TCN synaptosomes. The BK channels regulating [(3)H]D-aspartate release from primary afferent nerve terminals projecting into the TCN displayed limited sensitivity to iberiotoxin, whereas those expressed on cortical synaptosomes were highly sensitive to this toxin. BK channels did not appear to be present on GABAergic nerve terminals from the TCN since [(3)H]-γ-aminobutyric acid release in this model was unaffected by BK channel activators or blockers. Gene expression studies revealed expression levels of the α subunit in the TG that were only 31.2 ± 2.1% of those found in cortical tissues. The β4 subunit was the accessory subunit expressed most abundantly in both the cortex and TG. Levels of β1 and β2 were low in both these areas although β2 expression in the TG was higher than that found in the cortex. Immunocytochemistry experiments showed that co-localization of α and β4 subunits (the accessory subunit most abundantly expressed in both brain areas) was more common in TCN synaptosomes than in cortical synaptosomes. On the basis of these findings, it is reasonable to hypothesize that BK channels expressed on glutamatergic terminals in the TCN and cortex have distinct pharmacological profiles, which probably reflect different α and β subunit combinations. Channels in the cortex seem to be composed mainly of α subunits and to a lesser degree by α and β4 subunits, whereas in the TG the α + β4 combination seems to prevail (although α and/or α + β2 channels cannot be excluded). In light of the BK channels' selective control of excitatory transmission and their pharmacological diversity, their effects on primary glutamatergic afferents projecting to TCN represent a potential target for drug therapy of migraines and other types of orofacial pain.
authors.affiliation: Institute of Pharmacology, School of Medicine, Catholic University of the Sacred Heart, Largo F. Vito 1, 00168, Rome, Italy.
authors.forename: Irene
authors.initials: I
authors.lastname: Samengo
authors.forename: Diego
authors.initials: D
authors.lastname: Currò
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Barrese
authors.forename: Maurizio
authors.initials: M
authors.lastname: Taglialatela
authors.forename: Maria
authors.initials: M
authors.lastname: Martire
mesh_terms: D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D001562:Benzimidazoles / Q000494:pharmacology; D002540:Cerebral Cortex / Q000378:metabolism*; D018698:Glutamic Acid / Q000378:metabolism*; D007211:Indoles / Q000494:pharmacology; D051036:Large-Conductance Calcium-Activated Potassium Channels / Q000096:biosynthesis* / Q000187:drug effects / Q000502:physiology; D008297:Male; D010455:Peptides / Q000494:pharmacology; D021122:Protein Subunits / Q000378:metabolism; D017208:Rats, Wistar; D013574:Synaptosomes / Q000378:metabolism; D014275:Trigeminal Caudal Nucleus / Q000378:metabolism*; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
chemical_list: D001562:Benzimidazoles; D007211:Indoles; D051036:Large-Conductance Calcium-Activated Potassium Channels; D010455:Peptides; D021122:Protein Subunits; C086491:NS 1619; D001224:Aspartic Acid; D018698:Glutamic Acid; C048220:paxilline; D005680:gamma-Aminobutyric Acid; C064719:iberiotoxin