title: Clock Genes Regulate the Circadian Expression of Piezo1, TRPV4, Connexin26, and VNUT in an Ex Vivo Mouse Bladder Mucosa.
abstract: OBJECTIVE
  ClockΔ19/Δ19 mice is an experimental model mouse for nocturia (NOC). Using the bladder mucosa obtained from ClockΔ19/Δ19 mice, we investigated the gene expression rhythms of mechanosensory cation channels such as transient receptor potential cation channel subfamily V member 4 (TRPV4) and Piezo1, and main ATP release pathways including vesicular nucleotide transporter (VNUT) and Connexin26(Cx26), in addition to clock genes.
  
  
  METHODS
  Eight- to twelve-week-old male C57BL/6 mice (WT) and age- and sex-matched C57BL/6 ClockΔ19/Δ19 mice, which were bred under 12-h light/dark conditions for 2 weeks, were used. Gene expression rhythms and transcriptional regulation mechanisms in clock genes, mechanosensor, Cx26 and VNUT were measured in the mouse bladder mucosa, collected every 4 hours from WT and ClockΔ19/Δ19 mice using quantitative RT-PCR, a Western blot analysis, and ChIP assays.
  
  
  RESULTS
  WT mice showed circadian rhythms in clock genes as well as mechanosensor, Cx26 and VNUT. Their expression was low during the sleep phase. The results of ChIP assays showed Clock protein binding to the promotor regions and the transcriptional regulation of mechanosensor, Cx26 and VNUT. In contrast, all of these circadian expressions were disrupted in ClockΔ19/Δ19 mice. The gene expression of mechanosensor, Cx26 and VNUT was maintained at a higher level in spite of the sleep phase.
  
  
  CONCLUSIONS
  Mechanosensor, Cx26 and VNUT expressed with circadian rhythm in the mouse bladder mucosa. The disruption of circadian rhythms in these genes, induced by the abnormalities in clock genes, may be factors contributing to NOC because of hypersensitivity to bladder wall extension.
authors.affiliation: Department of Urology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Ihara
authors.affiliation: Department of Urology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Takahiko
authors.initials: T
authors.lastname: Mitsui
authors.affiliation: Department of Immunology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Nakamura
authors.affiliation: Department of Urology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Satoru
authors.initials: S
authors.lastname: Kira
authors.affiliation: Department of Urology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nakagomi
authors.affiliation: Department of Urology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Norifumi
authors.initials: N
authors.lastname: Sawada
authors.affiliation: Department of Neuropharmacology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Yuri
authors.initials: Y
authors.lastname: Hirayama
authors.affiliation: Department of Neuropharmacology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Shibata
authors.affiliation: Department of Neuropharmacology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Eiji
authors.initials: E
authors.lastname: Shigetomi
authors.affiliation: Department of Neuropharmacology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Shinozaki
authors.affiliation: Department of Urology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Mitsuharu
authors.initials: M
authors.lastname: Yoshiyama
authors.affiliation: Wake Forest University, Institute for Regenerative Medicine, Winston-Salem, North Carolina, United State of America.
authors.forename: Karl-Erik
authors.initials: KE
authors.lastname: Andersson
authors.affiliation: Department of Immunology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Atsuhito
authors.initials: A
authors.lastname: Nakao
authors.affiliation: Department of Urology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Takeda
authors.affiliation: Department of Neuropharmacology, Interdisciplinary Graduate School of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Schuichi
authors.identifier: http://orcid.org/0000-0001-6184-3106
authors.initials: S
authors.lastname: Koizumi
mesh_terms: D000818:Animals; D002940:Circadian Rhythm; D000072259:Connexin 26; D017630:Connexins / Q000235:genetics*; D004195:Disease Models, Animal; D005786:Gene Expression Regulation; D007473:Ion Channels / Q000235:genetics*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D053158:Nocturia / Q000235:genetics / Q000378:metabolism; D033704:Nucleotide Transport Proteins / Q000235:genetics*; D050916:TRPV Cation Channels / Q000235:genetics*; D001743:Urinary Bladder / Q000378:metabolism*
chemical_list: D017630:Connexins; C000607043:Gjb2 protein, mouse; D007473:Ion Channels; D033704:Nucleotide Transport Proteins; C552920:Piezo1 protein, mouse; C578984:Slc17a9 protein, mouse; D050916:TRPV Cation Channels; C486576:Trpv4 protein, mouse; D000072259:Connexin 26