title: Corticotrophin-releasing factor augments the I(H) in rat hypothalamic paraventricular nucleus parvocellular neurons in vitro.
abstract: The goal of this study was to characterize the effects of corticotrophin-releasing factor (CRF) on rat paraventricular nucleus (PVN) putative parvocellular neurons using whole cell patch-clamp recordings and single-cell reverse transcription-multiplex polymerase chain reaction (single-cell RT-mPCR) techniques. Under current clamp, CRF (10-600 nM) increased the neuronal basal firing rate and depolarized neurons in a dose-dependent manner. CRF-induced depolarization was unaffected by co-perfusion with TTX, 6-cyano-7-nitroquinoxaline-2 3-dione (CNQX), and bicuculline but was completely inhibited by ZD7288. Under voltage clamp, 300 nM CRF significantly increased the hyperpolarization-activated cation current (I(H)) in a voltage-dependent manner, shifted the I(H) conductance-voltage relationship (V 1/2) toward depolarization by approximately 7.8 mV, and enhanced the I(H) kinetics without changing the slope constant (k). Extracellular application of ZD7288 completely blocked I(H) and the CRF-induced increase in I(H). Furthermore, CRF-induced effects were completely blocked by extracellular application of 1 microM alpha-helical CRF-(9-14) (alpha-hCRF), a nonselective CRF receptor antagonist, but were not affected by extracellular application of antisauvagine-30, a selective CRF-receptor 2 antagonist. Single-cell RT-mPCR analysis showed that these neurons co-expressed CRF receptor 1 mRNA and CRF receptor 2 mRNA. Furthermore, CRF-sensitive neurons co-expressed HCN1 channel mRNA, HCN2 channel mRNA, and HCN3 channel mRNA, but not HCN4 channel mRNA. These results suggest that CRF modulates the subpopulation of PVN parvocellular neuronal function by CRF-receptor 1-mediated potentiation of HCN ion channel activity.
authors.affiliation: Department of Physiology, Miyazaki Medical College, University of Miyazaki, 5200 Kihara, Kiyotake-cho, Miyazaki-gun, Miyazaki 889-1692, Japan.
authors.forename: De-Lai
authors.initials: DL
authors.lastname: Qiu
authors.forename: Chun-Ping
authors.initials: CP
authors.lastname: Chu
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Shirasaka
authors.forename: Hiromasa
authors.initials: H
authors.lastname: Tsukino
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Nakao
authors.forename: Kazuo
authors.initials: K
authors.lastname: Kato
authors.forename: Takato
authors.initials: T
authors.lastname: Kunitake
authors.forename: Takahiko
authors.initials: T
authors.lastname: Katoh
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kannan
mesh_terms: D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D000831:Animals, Newborn; D015152:Blotting, Northern / Q000379:methods; D003346:Corticotropin-Releasing Hormone / Q000494:pharmacology*; D054815:Cyclic Nucleotide-Gated Cation Channels; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D006727:Hormone Antagonists / Q000494:pharmacology; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; D066298:In Vitro Techniques; D007473:Ion Channels / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D008297:Male; D009474:Neurons / Q000187:drug effects*; D010286:Paraventricular Hypothalamic Nucleus / Q000166:cytology*; D018408:Patch-Clamp Techniques / Q000379:methods; D010446:Peptide Fragments / Q000494:pharmacology; D015221:Potassium Channels; D011743:Pyrimidines / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar; D018019:Receptors, Corticotropin-Releasing Hormone / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
chemical_list: D054815:Cyclic Nucleotide-Gated Cation Channels; C578667:Hcn1 protein, rat; D006727:Hormone Antagonists; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; D007473:Ion Channels; D010446:Peptide Fragments; D015221:Potassium Channels; D011743:Pyrimidines; D012333:RNA, Messenger; D018019:Receptors, Corticotropin-Releasing Hormone; C082246:ICI D2788; D003346:Corticotropin-Releasing Hormone; C045089:corticotropin releasing hormone (9-41)