title: TGF-beta1 inhibits Cx43 expression and formation of functional syncytia in cultured smooth muscle cells from human detrusor.
abstract: BACKGROUND
  Human detrusor smooth muscle cells (hBSMCs) are coupled by connexin 43 (Cx43)-positive gap junctions to form functional syncytia. Gap junctional communication likely is necessary for synchronised detrusor contractions and is supposed to be altered in voiding disturbances. Other authors have shown that the pleiotropic cytokine TGF-beta1 upregulates Cx43 expression in human aortic smooth muscle cells.
  
  
  OBJECTIVE
  In this study, we examined the TGF-beta1 effects on Cx43 expression in cultured hBSMCs.
  
  
  METHODS
  hBSMC cultures, established from patients undergoing cystectomy, were treated with recombinant human TGF-beta1.
  
  
  METHODS
  Cx43 expression was then examined by Western blotting, real-time PCR, and immunocytochemistry. Dye-injection experiments were used to study the size of functional syncytia.
  
  
  CONCLUSIONS
  Dye-coupling experiments revealed stable formation of functional syncytia in passaged cell cultures (P1-P4). Stimulation with TGF-beta1 led to significant reduction of Cx43 immunoreactivity and coupling. Cx43 protein expression was significantly downregulated and Cx43 mRNA was only 30% of the control level. Interestingly, low phosphorylation species of Cx43 were particularly affected.
  
  
  CONCLUSIONS
  Our experiments demonstrated a significant down regulation of connexin 43 by TGF-beta1 in cultured hBSMCs. These findings support the view that TGF-beta1 is involved in the pathophysiology of urinary bladder dysfunction.
authors.affiliation: Department of Urology, University of Leipzig, Leipzig, Germany. jochen.neuhaus@medizin.uni-leipzig.de
authors.forename: Jochen
authors.initials: J
authors.lastname: Neuhaus
authors.forename: Marco
authors.initials: M
authors.lastname: Heinrich
authors.forename: Thilo
authors.initials: T
authors.lastname: Schwalenberg
authors.forename: Jens-Uwe
authors.initials: JU
authors.lastname: Stolzenburg
mesh_terms: D000368:Aged; D001011:Aorta / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D018031:Connexin 43 / Q000187:drug effects / Q000235:genetics*; D015536:Down-Regulation / Q000187:drug effects; D005260:Female; D017629:Gap Junctions / Q000187:drug effects / Q000502:physiology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007249:Inflammation / Q000235:genetics; D008297:Male; D009130:Muscle, Smooth / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D012334:RNA, Neoplasm / Q000235:genetics; D014158:Transcription, Genetic; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*; D001743:Urinary Bladder / Q000187:drug effects / Q000502:physiology / Q000503:physiopathology; D001749:Urinary Bladder Neoplasms / Q000235:genetics / Q000473:pathology* / Q000601:surgery*
chemical_list: D018031:Connexin 43; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D053773:Transforming Growth Factor beta1