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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Kruppel-like factor 5 is Required for Urothelial Maturation

ABSTRACT: Kruppel-like transcription factor 5 (Klf5) is expressed during late embryogenesis in the forming murine bladder urothelium. Targeted disruption of the Klf5flox alleles by the ShhGfpCre transgene resulted in failure of the bladder urothelium to mature accompanied by hydronephrosis, hydroureter, and vesicoureteric reflux in all E18.5 fetuses. The bladder urothelium did not stratify nor did it express terminal differentiation markers characteristic of basal, intermediate, and umbrella cells including keratins 20, 14, and 5, and uroplakins. At E18.5, an ectopic alpha smooth muscle actin positive layer of cells was identified subjacent to the undifferentiated Klf5-deficient urothelium. The effects of Klf5 deficiency were unique to the urothelium since maturation of the epithelium comprising the bladder neck and urethra were unaffected by the lack of KLF5. mRNA microarray analysis of whole E14.5 control and Klf5 deficient bladders identified Ppar? and Grhl3 as putative downstream intermediary transcription factors that regulate urothelial maturation. Transient transfection assays demonstrated that KLF5 regulated expression of the mGrhl3 promoter. These observations show that alterations in maturation of the bladder urothelium alone are sufficient to induce bladder dysfunction leading to prenatal hydronephrosis. Total RNA from E14.5 whole bladders isolated from Klf5D/D and Klf5+/+ShhGfpCre+ embryos (n=3 samples/genotype, each sample a pool of two bladders) was isolated using the Qiagen MicroRNA Kit. The cDNA was then hybridized according to the manufacturer’s protocol to the Affymetrix Mouse Gene 1.0 ST Array

ORGANISM(S): Mus musculus

SUBMITTER: Yan Xu

PROVIDER: E-GEOD-27014 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Kruppel-like factor 5 is Required for Urothelial Maturation

Project description:Kruppel-like transcription factor 5 (Klf5) is expressed during late embryogenesis in the forming murine bladder urothelium. Targeted disruption of the Klf5flox alleles by the ShhGfpCre transgene resulted in failure of the bladder urothelium to mature accompanied by hydronephrosis, hydroureter, and vesicoureteric reflux in all E18.5 fetuses. The bladder urothelium did not stratify nor did it express terminal differentiation markers characteristic of basal, intermediate, and umbrella cells including keratins 20, 14, and 5, and uroplakins. At E18.5, an ectopic alpha smooth muscle actin positive layer of cells was identified subjacent to the undifferentiated Klf5-deficient urothelium. The effects of Klf5 deficiency were unique to the urothelium since maturation of the epithelium comprising the bladder neck and urethra were unaffected by the lack of KLF5. mRNA microarray analysis of whole E14.5 control and Klf5 deficient bladders identified Ppar-gamma and Grhl3 as putative downstream intermediary transcription factors that regulate urothelial maturation. Transient transfection assays demonstrated that KLF5 regulated expression of the mGrhl3 promoter. These observations show that alterations in maturation of the bladder urothelium alone are sufficient to induce bladder dysfunction leading to prenatal hydronephrosis.

2011-08-02 | GSE27014 | GEO

Partial urethra obstruction in rat

Project description:Urethra was partially ligated and the urinary bladder was removed 10 days or 6 weeks after obstruction. Sham operated rats were used as controls. An addtitonal group of rats were repoerated 6 weeks after surgery and the obstruction was removed. These rats were then sacrificed 10 days after deobstruction. The bladder (including the urothelium) was frozen and used for RNA extraction. Urinary bladders from sham-operated, 10 days and 6 weeks after obstruction and rats 10 days after deobstruction were compared. mRNA and microRNA were both analyzed.

2013-11-24 | E-GEOD-47080 | biostudies-arrayexpress

Lrig2 is required for maturation of peripheral nerves in the urinary bladder

Project description:Leucine-rich-repeats and immunoglobulin-like-domains (LRIG2) variants can occur in urofacial syndrome (UFS), an inherited disease characterised by functional bladder outlet obstruction. We aimed to define the pathobiology underlying UFS which we hypothesised would illuminate how the bladder becomes innervated. Lrig2 was detected in pelvic ganglia supplying autonomic neurons to the bladder, and in neurites and glia emanating from explanted ganglia. One week old Lrig2 mutant mice displayed abnormally patterned bladder nerves, as did mice with mutations of Hpse2, also mutated in some UFS patients. From two weeks postnatally, Lrig2 mutants had urination defects resembling UFS. Molecules implicated in neural biology, including Nos1 which relaxes the bladder outlet, were dysregulated in newborn Lrig2 mutant bladders. These molecular aberrations preceded manifest urination defects. We discovered novel homozygous missense LRIG2 variants in non-syndromic bladder outlet obstruction. Molecules mutated in UFS are required for bladder innervation and LRIG2 variants can occur in non-syndromic bladder disease as well as in UFS.

2020-04-27 | E-MTAB-6089 | biostudies-arrayexpress

Time course for bladder UCC development in UPII-SV40Tag mice

Project description:We have identified genes that are differentially expressed between the bladders of UPII-SV40Tag mice and their age-matched wild-type littermates at 3, 6, 20, and 30 weeks of age. These are ages that correspond to premalignant, carcinoma in situ, and early-stage and later stage invasive UCC, respectively Analysis of the microarray data sets has revealed ∼1,900 unique genes differentially expressed (≥3-fold difference at one or more time points) between wild-type and UPII-SV40Tag urothelium during the time course of tumor development. RNA from the urothelium of duplicate UPII-SV40Tag hemizygous and age-matched wild type littermate mice was isolated at 3, 6, 20, and 30 weeks of age and gene expression profiles were obtained.

2010-08-28 | E-GEOD-23845 | biostudies-arrayexpress

Kruppel-like factor 5 is Required for Villus Formation and Terminal Differentiation of the Intestinal Epithelium

Project description:To identify putative KLF5 target genes that may mediate villus morphogenesis and epithelial maturation, we performed microarray analysis on intestinal samples isolated on E14.5, prior to the initiation of villus formation. Total RNA was isolated from dissected intestine segments located ~1.5 cm anterior to the cecum from E14.5 Klf5D/D and Klf5wt/wtShhEGFP/Cre+ embryos (n=3 samples/genotype, each sample a pool of two intestines). Gene expression profiles were compared using Affymetrix Mouse Gene 1.0 ST Array. Genes involved in the regulation of "epithelial cell differentiation/maturation”, “cell adhesion”, and “lipid synthesis, metabolism, and localization”. Consistent with the lack of terminally differentiated cell types in the intestines of E18.5 Klf5D/D fetuses. To identify putative KLF5 target genes that may mediate villus morphogenesis and epithelial maturation, we performed microarray analysis on intestinal samples isolated on E14.5, prior to the initiation of villus formation. Total RNA was isolated from dissected intestine segments located ~1.5 cm anterior to the cecum from E14.5 Klf5D/D and Klf5wt/wtShhEGFP/Cre+ embryos (n=3 samples/genotype, each sample a pool of two intestines). Gene expression profiles were compared using Affymetrix Mouse Gene 1.0 ST Array.

2013-01-24 | E-GEOD-39624 | biostudies-arrayexpress

Gene expression profiles of E13 bladder neck/urethral compartments (GUDMAP Series ID: 25)

Project description:The long term objective is to create an encyclopedia of the expression levels of all genes in multiple components of the developing bladder. The central thesis is straightforward. The combination of micro dissected tissues and FACS sorted cells plus microarray analysis offers a powerful, efficient and effective method for the creation of a global gene expression atlas of the developing urogenital system. Microarrays with essentially complete genome coverage can be used to quantitate expression levels of every gene. The ensuing rapid read-out provides an expression atlas that is more sensitive, more economical and more complete than would be possible by in situ hybridizations alone. The data submitted here delineates the gene expression profiles of the mesenchymal and epithelial compartments of the E13 mouse bladder neck/urethral compartment. FVB/N mice were time mated. At embryonic day 13 mice were euthanized by decapitation and the bladders were microdissected and cut just below the ureters. Since the EDTA dissociation was ineffective, the bladder neck and urethra were collected and treated with 1 mg/ml trypsin in Tyrode's solution for 20 minutes at 37°C. The layers were separated using a fine needle and rimming. The samples were placed in RLT and stored at -80°C. RNA was prepared and the Epicentre 2-round amplification scheme described under the Lessard Group Protocols on the GUDMAP pages were performed. The amplified RNA was examined with the Affymetrix GeneChip Mouse Genome 430 2.0 Array.

2009-04-24 | E-GEOD-15794 | biostudies-arrayexpress

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2018-02-09 | PXD007571 | Pride

Partial urethra obstruction in rat

2013-11-24 | GSE47080 | GEO

Gene activity induced by denervation in the rat urinary bladder [miRNA]

Project description:Bilateral freezing of the pelvic ganglia in female rats were performed to denervate the urinary bladder. Sham operated rats were used as controls. The rats were sacrificed 10 days after surgery. The urinary bladders (including the urothelium) were frozen and used for RNA extraction.

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Gene activity induced by denervation in the rat urinary bladder [mRNA]

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