title: rKv1.2 overexpression in the central medial thalamic area decreases caffeine-induced arousal.
abstract: The voltage-gated potassium channel Kv1.2 belongs to the shaker-related family and has recently been implicated in the control of sleep profile on the basis of clinical and experimental evidence in rodents. To further investigate whether increasing Kv1.2 activity would promote sleep occurrence in rats, we developed an adeno-associated viral vector that induces overexpression of rat Kv1.2 protein. The viral vector was first evaluated in vitro for its ability to overexpress rat Kv1.2 protein and to produce functional currents in infected U2OS cells. Next, the adeno-associated Kv1.2 vector was injected stereotaxically into the central medial thalamic area of rats and overexpression of Kv1.2 was showed by in situ hybridization, ex vivo electrophysiology and immunohistochemistry. Finally, the functional effect of Kv1.2 overexpression on sleep facilitation was investigated using telemetry system under normal conditions and following administration of the arousing agent caffeine, during the light phase. While no differences in sleep profile were observed between the control and the treated animals under normal conditions, a decrease in the pro-arousal effect of caffeine was seen only in the animals injected with the adeno-associated virus-Kv1.2 vector. Overall, our data further support a role of the Kv1.2 channel in the control of sleep profile, particularly under conditions of sleep disturbance.
authors.affiliation: Neurosciences Centre of Excellence for Drug Discovery, GlaxoSmithKline, Medicines Research Centre, Verona, Italy. cazzin-c@libero.it
authors.forename: C
authors.initials: C
authors.lastname: Cazzin
authors.forename: L
authors.initials: L
authors.lastname: Piccoli
authors.forename: M
authors.initials: M
authors.lastname: Massagrande
authors.forename: N
authors.initials: N
authors.lastname: Garbati
authors.forename: F
authors.initials: F
authors.lastname: Michielin
authors.forename: H-G
authors.initials: HG
authors.lastname: Knaus
authors.forename: C J A
authors.initials: CJ
authors.lastname: Ring
authors.forename: A D
authors.initials: AD
authors.lastname: Morrison
authors.forename: E
authors.initials: E
authors.lastname: Merlo-Pich
authors.forename: Z
authors.initials: Z
authors.lastname: Rovo
authors.forename: S
authors.initials: S
authors.lastname: Astori
authors.forename: A
authors.initials: A
authors.lastname: Lüthi
authors.forename: C
authors.initials: C
authors.lastname: Corti
authors.forename: M
authors.initials: M
authors.lastname: Corsi
mesh_terms: D000818:Animals; D001143:Arousal / Q000187:drug effects* / Q000235:genetics*; D001522:Behavior, Animal / Q000502:physiology; D002110:Caffeine / Q000494:pharmacology*; D002478:Cells, Cultured; D000697:Central Nervous System Stimulants / Q000494:pharmacology*; D000229:Dependovirus / Q000235:genetics; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D020646:Intralaminar Thalamic Nuclei / Q000378:metabolism*; D051663:Kv1.2 Potassium Channel / Q000235:genetics*; D008297:Male; D018408:Patch-Clamp Techniques; D051381:Rats; D012890:Sleep / Q000235:genetics / Q000502:physiology; D013686:Telemetry
chemical_list: D000697:Central Nervous System Stimulants; D051663:Kv1.2 Potassium Channel; D049452:Green Fluorescent Proteins; D002110:Caffeine