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Detrusor induction of miR-132/212 following bladder outlet obstruction: association with MeCP2 repression and cell viability.

Sadegh MK, Ekman M, Krawczyk K, Svensson D, Göransson O, Dahan D, Nilsson BO, Albinsson S, Uvelius B, Swärd K - PLoS ONE (2015)

Bottom Line: 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.Moreover, we found that serum stimulation and protein kinase C activation induced miR-132/212 in human detrusor cells.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Medical Science, Lund University, Lund, Sweden.

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.

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MiR-132/212 induction in the bladder correlates inversely with previously validated miR-132/212 targets.The normalized sum of miR-132 and miR-212 in bladders from sham-operated, obstructed (10 days and 6 weeks) and de-obstructed rats was correlated with the mRNA levels for Mecp2 (A), Ep300 (B), Pnkd (C), Jarid1a (D), Sh3bp5 (E), Ripk2 (F), Foxo3 (G), Pten (H) and Rasa1 (I). Expression data were obtained from microarray experiments (GEO accession GSE47080). Each symbol represents one rat and the 10d obstructed rats are represented by the rightmost symbols. The p-values obtained for the individual correlations are given at the bottom left in each diagram.
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pone.0116784.g004: MiR-132/212 induction in the bladder correlates inversely with previously validated miR-132/212 targets.The normalized sum of miR-132 and miR-212 in bladders from sham-operated, obstructed (10 days and 6 weeks) and de-obstructed rats was correlated with the mRNA levels for Mecp2 (A), Ep300 (B), Pnkd (C), Jarid1a (D), Sh3bp5 (E), Ripk2 (F), Foxo3 (G), Pten (H) and Rasa1 (I). Expression data were obtained from microarray experiments (GEO accession GSE47080). Each symbol represents one rat and the 10d obstructed rats are represented by the rightmost symbols. The p-values obtained for the individual correlations are given at the bottom left in each diagram.

Mentions: We next searched the literature for validated miR-132/212 targets and selected nine for which correlations with miR-132 and miR-212 were examined. Highly significant correlations in the expected directions were found for Mecp2, Ep300 and Pnkd (Fig. 4A-C). Correlations were significant also for Jarid1a, Sh3bp5, Ripk2 and Foxo3 (Fig. 4D-G). Pten and Rasa1 did not correlate with the miR-132/212 level (Fig. 4H-I). Ache was filtered due to its low expression level, and was not examined.


Detrusor induction of miR-132/212 following bladder outlet obstruction: association with MeCP2 repression and cell viability.

Sadegh MK, Ekman M, Krawczyk K, Svensson D, Göransson O, Dahan D, Nilsson BO, Albinsson S, Uvelius B, Swärd K - PLoS ONE (2015)

MiR-132/212 induction in the bladder correlates inversely with previously validated miR-132/212 targets.The normalized sum of miR-132 and miR-212 in bladders from sham-operated, obstructed (10 days and 6 weeks) and de-obstructed rats was correlated with the mRNA levels for Mecp2 (A), Ep300 (B), Pnkd (C), Jarid1a (D), Sh3bp5 (E), Ripk2 (F), Foxo3 (G), Pten (H) and Rasa1 (I). Expression data were obtained from microarray experiments (GEO accession GSE47080). Each symbol represents one rat and the 10d obstructed rats are represented by the rightmost symbols. The p-values obtained for the individual correlations are given at the bottom left in each diagram.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4305303&req=5

pone.0116784.g004: MiR-132/212 induction in the bladder correlates inversely with previously validated miR-132/212 targets.The normalized sum of miR-132 and miR-212 in bladders from sham-operated, obstructed (10 days and 6 weeks) and de-obstructed rats was correlated with the mRNA levels for Mecp2 (A), Ep300 (B), Pnkd (C), Jarid1a (D), Sh3bp5 (E), Ripk2 (F), Foxo3 (G), Pten (H) and Rasa1 (I). Expression data were obtained from microarray experiments (GEO accession GSE47080). Each symbol represents one rat and the 10d obstructed rats are represented by the rightmost symbols. The p-values obtained for the individual correlations are given at the bottom left in each diagram.
Mentions: We next searched the literature for validated miR-132/212 targets and selected nine for which correlations with miR-132 and miR-212 were examined. Highly significant correlations in the expected directions were found for Mecp2, Ep300 and Pnkd (Fig. 4A-C). Correlations were significant also for Jarid1a, Sh3bp5, Ripk2 and Foxo3 (Fig. 4D-G). Pten and Rasa1 did not correlate with the miR-132/212 level (Fig. 4H-I). Ache was filtered due to its low expression level, and was not examined.

Bottom Line: 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.Moreover, we found that serum stimulation and protein kinase C activation induced miR-132/212 in human detrusor cells.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Medical Science, Lund University, Lund, Sweden.

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.

Show MeSH
Related in: MedlinePlus