Identification of HCN4-expressing neurons using HCN4-luciferase knock-in mouse

• Project/Area Number: 26670292
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Pain science
• Research Institution: Kurume University
• Principal Investigator: TAKANO Makoto 久留米大学, 医学部, 教授 (30236252)
• Co-Investigator(Renkei-kenkyūsha): NAKASHIMA Noriyuki 久留米大学, 医学部, 助教 (80625468)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: HCN4 / ニューロン / 脊髄後角 / 大脳皮質 / 神経障害性疼痛 / 過分極誘発陽イオンチャネル / 脊髄 / 痛覚

Outline of Final Research Achievements

We generated transgenic mouse, in which cDNA of firefly luciferase-GFP fusion protein was knocked in at HCN4 locus (HCN4+/Luc), and visualized HCN4-expressing neurons in central nervous system. HCN4-expressing neurons were found in dorsal horn of spinal cord, and in III layer of sensory cortex.

Research Products

• [Journal Article] The hyperpolarization-activated cyclic nucleotide-gated (HCN) channels contain multiple S-palmitoylation sites (2016)
  • Author(s): Itoh, M., Ishihara, K., Nakashima, N., Takano, M.
  • Journal Title: J Physiol Sci Volume: 66 Issue: 3 Pages: 241-8
  • DOI: 10.1007/s12576-015-0420-5
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Clinical Concentrations of Local Anesthetics Bupivacaine and Lidocaine Differentially Inhibit Human Kir2.x Inward Rectifier K+ Channels (2016)
  • Author(s): Nakahira, K., Oshita, K., Itoh, M., Takano, M., Sakaguchi, Y., Ishihara, K.
  • Journal Title: Anesth Analg Volume: 122 Issue: 4 Pages: 1038-47
  • DOI: 10.1213/ane.0000000000001137
  • Peer Reviewed
• [Journal Article] Neuronal and glial expression of inward rectifier potassium channel subunits Kir2.x in rat dorsal root ganglion and spinal cord (2016)
  • Author(s): Murata Y, Yasaka T, Takano M, Ishihara K
  • Journal Title: Neuroscience Letters Volume: 617 Pages: 59-65
  • DOI: 10.1016/j.neulet.2016.02.007
  • Peer Reviewed
• [Journal Article] Renal Nerve-Mediated Erythropoietin Release Confers Cardioprotection During Remote Ischemic Preconditioning (2015)
  • Author(s): Oba T, Yasukawa H, Nagata T, Kyogoku S, Minami T, Nishihara M, Ohshima H, Mawatari K, Nohara S, Takahashi J, Sugi S, Igata S, Iwamoto Y, Kai H, Matsuoka H, Takano M, Aoki H, Fukumoto Y, Imaizumi I.
  • Journal Title: Circulation Journal Volume: 79 Issue: 7 Pages: 1557-1567
  • DOI: 10.1253/circj.CJ-14-1171
  • NAID: 130005083944
  • ISSN: 1346-9843, 1347-4820
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Genotype-dependent differences in age of manifestation and arrhythmia complications in short QT syndrome. (2015)
  • Author(s): Harrell DT, Ashihara T, Ishikawa T, et al.
  • Journal Title: International Journal of Cardiology. Volume: 190 Pages: 393-402
  • DOI: 10.1016/j.ijcard.2015.04.090
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] ペースメーカーチャネル(HCN)の病態生理. (2015)
  • Author(s): 鷹野 誠
  • Journal Title: 臨と研 Volume: 92 Pages: 134-137
• [Journal Article] Disturbance of cardiac gene expression and cardiomyocyte structure predisposes Mecp2-null mice to arrhythmias. (2015)
  • Author(s): Hara M, Takahashi T, Mistumasu C, Igata S, Takano M, Minami T, Yasukawa H, Okayama S, Nakamura K, Okabe Y, Tanaka E, Takemura G, Kosai K, Yamashita Y, Matsuishi T.
  • Journal Title: Sci Rep Volume: 5 Issue: 1 Pages: 11204-11204
  • DOI: 10.1038/srep11204
  • Peer Reviewed / Open Access
• [Journal Article] Enhancement of Spontaneous Activity by HCN4 overexpression in Mouse Embryonic Stem Cell-derived Cardiomyocytes - a Possible Biological Pacemaker. (2015)
  • Author(s): SaitoY, Nakamura K, Yoshida M, Sugiyama H, Ohe T, Kurokawa J, Furukawa T, Takano M, Nagase S, Morita H, Kusano KF, Ito H.
  • Journal Title: PLoS ONE Volume: 10 Issue: 9 Pages: e0138193-e0138193
  • DOI: 10.1371/journal.pone.0138193
  • Peer Reviewed / Open Access
• [Journal Article] Ectopic automaticity induced in ventricular myocytes by transgenic overexpression of HCN2. (2015)
  • Author(s): Oshita K, Itoh M, Hirashima S, Kuwabara Y, Ishihara K, Kuwahara K, Nakao K, Kimura T, Nakamura K, Ushijima K, Takano M.
  • Journal Title: Journal of Molecular and Cellular Cardiology Volume: 80 Pages: 81-89
  • DOI: 10.1016/j.yjmcc.2014.12.019
  • NAID: 120007100131
  • Peer Reviewed / Open Access
• [Journal Article] Mechanism of the complete block of the Kir2.1 inward rectifier K+ channel under the external K+-free condition. (2015)
  • Author(s): Yanagi-Ishihara K, Itoh M, Takano M
  • Journal Title: The Journal of Physiological Sciences Volume: 65
• [Journal Article] Zdhhc3/7, the members of protein-palmitoylation enzymes, inhibit the current amplitude of HCN2 channel. (2015)
  • Author(s): Itoh M, Ishihara K, Takano M
  • Journal Title: The Journal of Physiological Sciences Volume: 65
• [Journal Article] Luminescence imaging of HCN4 expression and phenotypic analysis of HCN4 TET-off mouse. (2015)
  • Author(s): Kozasa Y, Ohshita K, Nakashima N, Ushijima K, Takano M.
  • Journal Title: The Journal of Physiological Sciences Volume: 65
• [Journal Article] Epithelium-dependent periodical excitation in response to stretch of guinea pig seminal vesicle. (2015)
  • Author(s): Takeya M, Hayashi T, Nakamura K, Takano M
  • Journal Title: The Journal of Physiological Sciences Volume: 65
• [Journal Article] 過分極誘発陽イオンチャネル（HCN2）の潜在的な催不整脈性. (2014)
  • Author(s): 大下健輔, 伊藤政之, 井形幸代, 桒原佳宏, 桑原宏一郎, 牛島一男, 鷹野 誠
  • Journal Title: 日本生理学雑誌 Volume: 76 Pages: 83-83
• [Journal Article] QT短縮症候群に同定されたK チャネル変異の電気生理学的特性と臨床像. (2014)
  • Author(s): ダニエル・トシオ・ハーレル, 芦原貴司, 冨永伊知子, 阿部圭祐, 石川泰輔, 住友直方, 鵜野起久也, 鷹野 誠, 蒔田直昌
  • Journal Title: 日本生理学雑誌 Volume: 76 Pages: 79-79
• [Presentation] HCN4過剰発現・ノックアウトマウスにおける洞房結節機能の変化. (2016)
  • Author(s): 鷹野 誠, 小佐々優子, 武谷三恵, 中島則行, 石原圭子
  • Organizer: 第55回日本生体医工学会大会
  • Place of Presentation: 富山国際会議場(富山市）
  • Year and Date: 2016-04-26
• [Presentation] Role of mucosa in generating spontaneous activity in the guinea pig seminal vesicle. (2016)
  • Author(s): Takeya M, Hashitani H, Hayashi T, Nakamura K, Takano M
  • Organizer: The 93rd Annual Meeting of the Physiological Society of Japan
  • Place of Presentation: Sapporo Convention Center(札幌）
  • Year and Date: 2016-03-22
• [Presentation] Revisiting K+ dependence of conductance and gating of strong inward rectifier K+ channels. (2016)
  • Author(s): Ishihara K, Itoh M, Igata S, Takano M.
  • Organizer: The 93rd Annual Meeting of the Physiological Society of Japan
  • Place of Presentation: Sapporo Convention Center(札幌）
  • Year and Date: 2016-03-22
• [Presentation] The role of pacemaker channel HCN4 against bradycardic response. (2016)
  • Author(s): Kozasa Y, Takeya M, Nakashima N, Ushijima K, Takano M.
  • Organizer: The 93rd Annual Meeting of the Physiological Society of Japan
  • Place of Presentation: Sapporo Convention Center(札幌）
  • Year and Date: 2016-03-22
• [Presentation] Visualization of HCN4-expressing neurons with GFP using Tet-expression system. (2016)
  • Author(s): Nakashima N, Takano M
  • Organizer: The 93rd Annual Meeting of the Physiological Society of Japan
  • Place of Presentation: Sapporo Convention Center(札幌）
  • Year and Date: 2016-03-22
• [Presentation] Arrhythmogenic properties of rat pulmonary vein cardiomyocytes. (2016)
  • Author(s): Okamoto Y, Takano M, Ono K.
  • Organizer: The 93rd Annual Meeting of the Physiological Society of Japan
  • Place of Presentation: Sapporo Convention Center(札幌）
  • Year and Date: 2016-03-22
• [Presentation] ペースメーカーチャネルHCN4の陰性変時作用における新たな機能. (2015)
  • Author(s): 小佐々優子, 武谷三恵, 中島則行, 牛島一男, 鷹野 誠
  • Organizer: 第66回西日本生理学会
  • Place of Presentation: 久留米大学筑水会館(久留米市）
  • Year and Date: 2015-10-09
• [Presentation] ノックインマウスを使ったHCN4 発現ニューロンの可視化. (2015)
  • Author(s): 中島則行, 鷹野 誠
  • Organizer: 第66回西日本生理学会
  • Place of Presentation: 久留米大学筑水会館(久留米市）
  • Year and Date: 2015-10-09
• [Presentation] 内向き整流性カリウムチャネルのポアの開口には細胞外カリウムイオンの存在を必要としない. (2015)
  • Author(s): 柳（石原）圭子, 井形幸代, 伊藤政之, 鷹野 誠
  • Organizer: 平成27年度生理学研究所研究会「膜システムの機能的・構造的統合」
  • Place of Presentation: 生理学研究所(岡崎）
  • Year and Date: 2015-09-01
• [Presentation] モルモット精嚢平滑筋自動性における粘膜の役割. (2015)
  • Author(s): 武谷三恵, 橋谷 光, 林 篤正, 中村桂一郎, 鷹野 誠
  • Organizer: 第57回日本平滑筋学会総会
  • Place of Presentation: 山口大学小串キャンパス（医学部）総合研究棟(山口）
  • Year and Date: 2015-08-25
• [Presentation] Controversy still goes on; lessons from HCN4 knock-out mice. (2015)
  • Author(s): Takano M, Kozasa Y, Takeya M, Ohshita K, Ito M, Ishihara K, Nakashima N.
  • Organizer: The Joint Meeting of the 30th Annual Meeting of the Japanese Heart Rhythm Society and the 32nd Annual Scientific Meeting of the Japanese Society of Electrocardiology
  • Place of Presentation: Kyoto International Conference Center(京都）
  • Year and Date: 2015-07-28
• [Presentation] Atrial fibrillation and arrhythmogenic activities of pulmonary vein cardiomyocytes. (2015)
  • Author(s): Ono K, Okamoto Y, Umehara S, Adachi T, Ohba T, Amano A, Takano M
  • Organizer: The Joint Meeting of the 30th Annual Meeting of the Japanese Heart Rhythm Society and the 32nd Annual Scientific Meeting of the Japanese Society of Electrocardiology
  • Place of Presentation: Kyoto International Conference Center(京都）
  • Year and Date: 2015-07-28
• [Presentation] Luminescence imaging of HCN4 expression and phenotypic analysis of HCN4 TET-off mouse. (2015)
  • Author(s): Kozasa Y, Ohshita K, Nakashima N, Ushijima K, Takano M.
  • Organizer: The 92nd Annual Meeting of the Physiological Society of Japan
  • Place of Presentation: 神戸コンベンションセンター（兵庫県神戸市）
  • Year and Date: 2015-03-22
• [Presentation] Epithelium-dependent periodical excitation in response to stretch of guinea pig seminal vesicle. (2015)
  • Author(s): Takeya M, Hayashi T, Nakamura K, Takano M.
  • Organizer: The 92nd Annual Meeting of the Physiological Society of Japan
  • Place of Presentation: 神戸コンベンションセンター（兵庫県神戸市）
  • Year and Date: 2015-03-22
• [Presentation] Mechanism of the complete block of the Kir2.1 inward rectifier K+ channel under the external K+-free condition. (2015)
  • Author(s): Yanagi-Ishihara K, Itoh M, Takano M.
  • Organizer: The 92nd Annual Meeting of the Physiological Society of Japan
  • Place of Presentation: 神戸コンベンションセンター（兵庫県神戸市）
  • Year and Date: 2015-03-21
• [Presentation] Zdhhc3/7, the members of protein-palmitoylation enzymes, inhibit the current amplitude of HCN2 channel. (2015)
  • Author(s): Itoh M, Ishihara K, Takano M.
  • Organizer: The 92nd Annual Meeting of the Physiological Society of Japan
  • Place of Presentation: 神戸コンベンションセンター（兵庫県神戸市）
  • Year and Date: 2015-03-21
• [Presentation] 化学発光によるHCN4発現部位の可視化 (2014)
  • Author(s): 小佐々優子
  • Organizer: 第65回西日本生理学会
  • Place of Presentation: 琉球大学(沖縄県中頭郡西原町）
  • Year and Date: 2014-10-24
• [Presentation] Differential expression of Kir2.x inward rectifier K+ channels in neurons and glial cells in rat dorsal root ganglion and spinal cord. (2014)
  • Author(s): Murata Y, Honda Y, Yasaka T, Takano M, Masuko S, Ishihara K.
  • Organizer: The 37th Annual Meeting of the Japan Neuroscience Society
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-09-11
• [Presentation] Analysis of cardiac arrhythmias and gene expression in MeCP2-null Mouse. (2014)
  • Author(s): Hara M, Takahashi T, Mitsumasu C, Igata S, Takano M, Okabe Y, Tanaka E, Matsuishi T.
  • Organizer: 13th Rett Syndrome Symposium
  • Place of Presentation: Chantilliy, VA, USA
  • Year and Date: 2014-06-24 – 2014-06-26
• [Remarks] 久留米大学研究者紹介
  • URL: http://research.kurume-u.ac.jp/data.php?scode=14905632890988
• [Remarks] 久留米大学研究者紹介
  • URL: http://research.kurume-u.ac.jp/data.php?scode=76735632890988
• [Remarks] 久留米大学医学部生理学講座統合自律機能部門研究業績
  • URL: http://www.med.kurume-u.ac.jp/med/physiol2/paper.html