DOI: 10.5061/dryad.3n5tb2rxg
title: Data from: Hedgehog pathway disruption alters urothelial differentiation and continence mechanisms in the female mouse lower urinary tract

description: Urinary incontinence (UI) imposes a significant healthcare burden and
 reduces quality of life. Contributing factors such as aging,
 pregnancy/childbirth, stress, and injury are recognized, but an incomplete
 understanding of underlying mechanisms limits new therapies. Hedgehog (Hh)
 signaling has been implicated in lower urinary tract development, but its
 specific role in female continence mechanisms has not been fully
 characterized. Here, we investigate the functional and molecular
 consequences of reduced Hh signaling using Gli2+/-;Gli3Δ699/+, (Gli
 mutant) female mice. We assessed spontaneous voiding through void spot
 assays and uroflowmetry, then assessed contractility in bladder and
 urethral tissues ex vivo. Female Gli mutant mice display more small-volume
 voids than wild-type mice. Gli mutant female bladders had reduced
 contractile strength to electrical field and cholinergic stimuli, while
 the urethra had reduced sensitivity to serotonin-mediated contraction, but
 not to phenylephrine. Thus, unique changes to bladder and urethral
 contractility dynamics are present in Gli mutant mice and are dependent
 upon the type of stimuli. Further, expression of serotonin transporter
 (Sert) mRNA was increased in the Gli mutant urethra compared to
 the wild type. Uroplakin IIIa, typically localized to bladder urothelium,
 was ectopically expressed in urethral urothelium of adult but not
 embryonic (E) day 16 Gli mutant mice. These findings highlight a
 previously uncharacterized role of Hh signaling in maintaining female
 lower urinary tract function and urothelial patterning, and support
 further investigation of its contribution to continence.
description: # Data from: Hedgehog pathway disruption alters urothelial differentiation
 and continence mechanisms in the female mouse lower urinary tract Dataset
 DOI: [10.5061/dryad.3n5tb2rxg](https://doi.org/10.5061/dryad.3n5tb2rxg) ##
 Description of the data and file structure Excel files of raw data for
 physiology and histology assessments in wild-type and Gli mutant female
 mice. ### Files and variables #### File: VSA_raw_data.xlsx
 **Description:** Raw data generated from running void spot assays on
 wild-type and mutant animals. After the voiding assay, the filter papers
 containing urine were imaged with UV light,t and the resulting images were
 analyzed via Void Whizzard software freely available as a plugin in Image
 J. This Excel table is the readout of the Void Whizzard analysis of the
 urine content on the assay papers. The first column is the animal's
 unique ID, the second column is the genotype of that animal the third
 column is the name of the void paper image file. Each column after that is
 the voiding parameter examined, and each row is a different animal. #####
 Variables * The first column is the animal's unique ID, the second
 column is the genotype of that animal the third column is the name of the
 void paper image file. Each column after that is the voiding parameter
 examined, and each row is a different animal. #### File:
 Uroflow_raw_data.xlsx **Description:** Raw data generated from running
 uroflowmetry on wild-type and mutant animals. After the assay, the videos
 captured by Raspberry Pi units during uroflowmetry testing were analyzed.
 ThisExcell table is the analysis of each captured void in the mass and
 time of each void. The first column is the animal's unique ID, and
 the second column is the genotype of that animal. Each column header is a
 parameter examined, and each row is a void event for each different
 animal. Columns U through AB are the averages of all parameters from all
 voids from a single animal. ##### Variables * The first column is
 theanimal'sl unique ID, the second column is the genotype of that
 animal. All column headers describe the variable assessed; each row is a
 void event split by each individual animal. Column U through AB are the
 averages of all parameters from all voids from a single animal. #### File:
 Bladder_bath_raw_data.xlsx **Description:** Raw data generated from
 bladder contractility assays on wild-type and mutant animals using
 LabChart software during testing. Each column header is a parameter
 examined, and each row is a different bladder separated for each animal.
 Two bladder halves were averaged to have one end value for each parameter
 from each animal.  ##### Variables * The first column is the animal's
 unique ID, and the second column is the genotype of that animal. All other
 columns are parameters measured. Row 1 names the type of analysis. Row 4
 and 5 indicate which treatment was applied, columns E-AD are the raw
 values of the parameter. All subsequent columns are calculations of the
 raw data in E-AD, as stated in Row 1 for that column. #### File:
 Urethra_bath_raw_data.xlsx **Description:** Raw data generated from
 urethral contractility assays on wild-type and mutant animals using
 LabChart software during wire myography testing. Each column header is a
 parameter examined, and each row is a different animal. ##### Variables *
 The first column is theanimal'sl unique ID, the second column is the
 sex, and the third column is the genotype of that animal. All other
 columns are parameters measured. Row 1 names the type of analysis. Row 2
 and 3 indicate which treatment was applied, columns D-AK are the raw
 values of the parameter. All subsequent columns are calculations off of
 the raw data in D-AK, and as stated in Row 1 for that column. #### File:
 Urethra_qPCR_raw_data.xlsx **Description:** Raw data generated from qPCR
 analysis of the urethra from wild-type and mutant mice. Each column header
 is a gene examined, and each row is a different animal. Raw Ct values are
 presented,d and the delta Ct calculation is presented. ##### Variables *
 The first column is the animal's unique ID, the second column is the
 genotype of that animal, third column is sex. All other columns are
 parameters measured. Columns D-J are raw Ct values. Column P-U is the
 calculation of the delta Ct using housekeeping gene Ppia for relative
 expression. #### File: Adult_IHC_raw_data.xlsx **Description:** Raw data
 generated from quantifying the percentage of total positive cells from
 images. Each column header is an endpoint examined, and each row is a
 different image, separated out for each animal. ##### Variables * All
 column headers describe the protein marker expressed, the cell count, or
 average; each row is an image, which is separated out for each animal.
 Each tab is a set of slides and stains used for cell counting. The summary
 tab is the average from all of those stains, and finally averaged for a
 single number for each animal. #### File: E16_IHC_raw_data.xlsx
 **Description:** Raw data generated from quantifyingthe percent total
 positive cells from images. Each column header is an endpoint examined,
 and each row is a different image, separated out for each animal. #####
 Variables * All column headers describe the protein marker expressed, cell
 count, or average, each row is an image, which is separated out for each
 animal. #### File: Body_and_tissue_mass_raw_data.xlsx **Description:** Raw
 data generated from all animals used in the study. Each column header is
 an endpoint examined, and each row is a different animal. ##### Variables
 * All column headers describe the endpoint assessed; each row is a
 different animal. #### File: Craniofacial_E16_Morphemetrics_raw_data.xlsx
 **Description:** Raw data generated from all animals used in the study.
 Each column header is an endpoint examined, and each row is a different
 animal. ##### Variables * All column headers describe the endpoint
 assessed; each row is a different animal. #### File: raw_vsa_images.zip
 **Description:** Images of papers from void spot assay (VSA) imaged under
 UV light from all animals used in the study.   ##### Variables * All file
 names are the animal ID. All are Tiffs of the imaged paper that was placed
 in the cage for the mouse to urinate on for 4 hours. #### File:
 e16_images.zip **Description:** Fluorescent images from embryonic day
 (E16) urogenital sinus tissue sections generated from all animals used in
 the study, which were stained via immunohistochemistry with antibodies
 targeting keratin 5 (red) and uroplakin IIIa (green), DAPI (nuclei, blue).
 All files are the merged red, green, and blue channels as a TIFF.  #####
 Variables * All file names describe the endpoint assessed and animal ID.
 File names are the block number, animal number, and a description of the
 antibodies used and their color, along with where in the urogenital sinus
 the image was taken. #### File: Krt5_Krt13_images.zip
 **Description:** Fluorescent images from bladder and urethra tissue
 sections generated from all animals used in the study, which were stained
 via immunohistochemistry with antibodies targeting keratin 5 (red) and
 keratin 13 (green), DAPI (nuclei, blue). All files are the merged red,
 green, and blue channels as a TIFF.  ##### Variables * All file names
 describe the endpoint assessed and animal ID. File names are the block
 number, animal number,r and a description of the antibodies used and their
 color, along with where in the tissue the image was taken. #### File:
 Krt5_Upk3a_images.zip **Description:** Fluorescent images from bladder and
 urethra tissue sections generated from all animals used in the study,
 which were stained via immunohistochemistry with antibodies targeting
 keratin 5 (red) and uroplakin IIIa (green), DAPI (nuclei, blue). All files
 are the merged red, green, and blue channels as a TIFF.  ##### Variables *
 All file names describe the endpoint assessed and animal ID. File names
 are the block number, animal number, and a description of the antibodies
 used and their color,r along with where in the tissue the image was taken.
 **Code/Software** All data can be viewedin Excell or a photo viewer. Void
 Whizzard software (free plugin for Image J) - was used for VSA analysis
 **Access information** Other publicly accessible locations of the data: *
 None Data was derived from the following sources: * None