title: Detrusor induction of miR-132/212 following bladder outlet obstruction: association with MeCP2 repression and cell viability.
abstract: The microRNAs (miRNAs) miR-132 and miR-212 have been found to regulate synaptic plasticity and cholinergic signaling and recent work has demonstrated roles outside of the CNS, including in smooth muscle. Here, we examined if miR-132 and miR-212 are induced in the urinary bladder following outlet obstruction and whether this correlates with effects on gene expression and cell growth. Three to seven-fold induction of miR-132/212 was found at 10 days of obstruction and this was selective for the detrusor layer. We cross-referenced putative binding sites in the miR-132/212 promoter with transcription factors that were predicted to be active in the obstruction model. This suggested involvement of Creb and Ahr in miR-132/212 induction. Creb phosphorylation (S-133) was not increased, but the number of Ahr positive nuclei increased. Moreover, we found that serum stimulation and protein kinase C activation induced miR-132/212 in human detrusor cells. To identify miR-132/212 targets, we correlated the mRNA levels of validated targets with the miRNA levels. Significant correlations between miR-132/212 and MeCP2, Ep300, Pnkd and Jarid1a were observed, and the protein levels of MeCP2, Pnkd and Ache were reduced after obstruction. Reduction of Ache however closely matched a 90% reduction of synapse density arguing that its repression was unrelated to miR-132/212 induction. Importantly, transfection of antimirs and mimics in cultured detrusor cells increased and decreased, respectively, the number of cells and led to changes in MeCP2 expression. In all, these findings show that obstruction of the urethra increases miR-132 and miR-212 in the detrusor and suggests that this influences gene expression and limits cell growth.
authors.affiliation: Department of Experimental Medical Science, Lund University, Lund, Sweden.
authors.forename: Mardjaneh Karbalaei
authors.initials: MK
authors.lastname: Sadegh
authors.affiliation: Department of Experimental Medical Science, Lund University, Lund, Sweden; Department of Biology, Lund University, Lund, Sweden.
authors.forename: Mari
authors.initials: M
authors.lastname: Ekman
authors.affiliation: Department of Experimental Medical Science, Lund University, Lund, Sweden.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Krawczyk
authors.affiliation: Department of Experimental Medical Science, Lund University, Lund, Sweden.
authors.forename: Daniel
authors.initials: D
authors.lastname: Svensson
authors.affiliation: Department of Experimental Medical Science, Lund University, Lund, Sweden.
authors.forename: Olga
authors.initials: O
authors.lastname: Göransson
authors.affiliation: Department of Experimental Medical Science, Lund University, Lund, Sweden.
authors.forename: Diana
authors.initials: D
authors.lastname: Dahan
authors.affiliation: Department of Experimental Medical Science, Lund University, Lund, Sweden.
authors.forename: Bengt-Olof
authors.initials: BO
authors.lastname: Nilsson
authors.affiliation: Department of Experimental Medical Science, Lund University, Lund, Sweden.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Albinsson
authors.affiliation: Department of Clinical Sciences, Lund University, Lund, Sweden.
authors.forename: Bengt
authors.initials: B
authors.lastname: Uvelius
authors.affiliation: Department of Experimental Medical Science, Lund University, Lund, Sweden.
authors.forename: Karl
authors.initials: K
authors.lastname: Swärd
mesh_terms: D000110:Acetylcholinesterase / Q000378:metabolism; D000818:Animals; D002470:Cell Survival / Q000235:genetics; D005260:Female; D058851:GPI-Linked Proteins / Q000378:metabolism; D006801:Humans; D051783:Methyl-CpG-Binding Protein 2 / Q000172:deficiency / Q000235:genetics*; D035683:MicroRNAs / Q000235:genetics*; D009124:Muscle Proteins / Q000378:metabolism; D009130:Muscle, Smooth / Q000378:metabolism* / Q000473:pathology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D018336:Receptors, Aryl Hydrocarbon / Q000378:metabolism; D013569:Synapses / Q000378:metabolism; D001748:Urinary Bladder Neck Obstruction / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*
chemical_list: D058851:GPI-Linked Proteins; C547077:MIRN132 microRNA, human; C551411:MIRN212 microRNA, human; D051783:Methyl-CpG-Binding Protein 2; D035683:MicroRNAs; D009124:Muscle Proteins; C495444:PNKD protein, human; D012333:RNA, Messenger; D018336:Receptors, Aryl Hydrocarbon; C548244:ACHE protein, human; D000110:Acetylcholinesterase