title: Changes of HCN gene expression and I(f) currents in Nkx2.5-positive cardiomyocytes derived from murine embryonic stem cells during differentiation.
abstract: Changes in the expression of hyperpolarization-activated cyclic nucleotide (HCN)-gated channels and I(f) currents during the differentiation of embryonic stem cells into cardiac cells remain unknown. We examined changes of HCN genes in expression and function during the differentiation of Nkx2.5-positive cardiac precursor cells derived from mouse ES cells using cell sorting, RTPCR, immunofluorescence and whole cell patch-clamp techniques. Cs(+)-induced inhibition of automaticity and transcription of HCN genes increased during differentiation. Expressions of Nkx2.5, a marker of cardiac progenitor cell, and Flk1, a marker of hemangioblast, were mutually exclusive. Messenger RNA and proteins encoded by HCN1 and 4 genes were predominantly observed in Nkx2.5-positive cells on day 15, although Flk1-positive cells did not express genes of the HCN family on that day. Cs(+)-induced prolongation of the cycle of spontaneous action potentials and I(f) currents were predominantly observed on day 15. These results suggested that a fraction of Nkx2.5-positive cardiac precursor cells was committed to pacemaking cells expressing I(f) channels predominantly encoded by HCN 1 and 4 genes.
authors: Department of Cardiovascular Medicine, Tottori University Faculty of Medicine, Japan.
authors: Shuichi
authors: S
authors: Yano
authors: Junichiro
authors: J
authors: Miake
authors: Einosuke
authors: E
authors: Mizuta
authors: Kasumi
authors: K
authors: Manabe
authors: Udin
authors: U
authors: Bahrudin
authors: Kumi
authors: K
authors: Morikawa
authors: Keita
authors: K
authors: Arakawa
authors: Norihito
authors: N
authors: Sasaki
authors: Osamu
authors: O
authors: Igawa
authors: Chiaki
authors: C
authors: Shigemasa
authors: Yasutaka
authors: Y
authors: Yamamoto
authors: Takayuki
authors: T
authors: Morisaki
authors: Kyoko
authors: K
authors: Hidaka
authors: Yasutaka
authors: Y
authors: Kurata
authors: Akio
authors: A
authors: Yoshida
authors: Goshi
authors: G
authors: Shiota
authors: Katsumi
authors: K
authors: Higaki
authors: Haruaki
authors: H
authors: Ninomiya
authors: Jong-Kook
authors: JK
authors: Lee
authors: Yasuaki
authors: Y
authors: Shirayoshi
authors: Ichiro
authors: I
authors: Hisatome
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D002469:Cell Separation; D002586:Cesium / Q000378:metabolism; D054815:Cyclic Nucleotide-Gated Cation Channels / Q000235:genetics / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology / Q000502:physiology*; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism
chemical_list: D054815:Cyclic Nucleotide-Gated Cation Channels; C578665:HCN1 protein, human; C578666:Hcn1 protein, mouse; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; C484469:Nkx2-5 protein, mouse; D015221:Potassium Channels; D012333:RNA, Messenger; D014157:Transcription Factors; D002586:Cesium; D040301:Vascular Endothelial Growth Factor Receptor-2