Doxycycline-responsive transcriptome in the mouse inner medullary collecting duct cell line (mIMCD3)
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ABSTRACT:
PROVIDER: PRJNA720114 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR14154101_1.fastq.gz | Fastqsanger.gz | |||
| SRR14154101_2.fastq.gz | Fastqsanger.gz | |||
| SRR14154102_1.fastq.gz | Fastqsanger.gz | |||
| SRR14154102_2.fastq.gz | Fastqsanger.gz | |||
| SRR14154103_1.fastq.gz | Fastqsanger.gz |
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Project description:Purpose: The goal of this study is to identify doxycycle-responsive genes in mouse kidney inner medullary collecting duct cell line mIMCD3. To explore compreshensive profile of doxycycline-mediated gene expression, transcriptomes of doxycycline-responsive genes at two different time points (3 days and 6 days) were profiled and analyzed. Methods: Total RNAs were isolated from mIMCD3 cells treated with doxycycline or vehicle at different time points (3 days and 6 days). Four replicates for vehicle- or doxycycline-treated group were generated at each tested time point. cDNA libraries were prepared using a Nextera DNA library preparation protocol. The sequence reads from Illumina HiSeq3000 platform were qualified and quantified at the transcript level using Salmon (0.14.1). Differential expression analysis were performed using edgeR. Results: mRNA profiles of mouse kidney inner medullary collecting duct mIMCD3 cells treated with doxycycline or vehicle (DMSO) for 3 days and 6 days were generated using an optimized RNA-Seq workflow. Transcript level quantification using a pseudo-alignment quantification method (Salmon) was performed to calculate transcript abundance in each sample. Then differential expression analysis identified the differentially expressed genes at each time point comparison (DOX vs vehicle). Conclusions: This study revealed comprehensive transcriptomic changes of doxycycline-responsive gene expression in mouse kidney inner medullary collecting duct cells.
2021-11-17 | GSE171573 | GEO
Project description:Identification of gene expressed in the enriched inner medullary collecting duct cells in rat. Experiment Overall Design: Rat inner medullary collecting duct (IMCD) cells were isolated from 7 male Sprage-Dawley rats by collagenase and hyaluronidase digestion and follow by low speed centrifugation. The non-IMCD cells were collected by centrifugation of supernatant of enriched IMCD samples. Experiment Overall Design: Total RNA about 3 ug were used per microarray (Rat 230 2.0 Genechip array). Experiment Overall Design: The experiments were repeat 3 times (3 pairs of IMCD VS non-IMCD)
2008-06-15 | E-GEOD-7891 | biostudies-arrayexpress
Project description:Identification of gene expressed in the enriched inner medullary collecting duct cells in rat. Keywords: gene expression comparision between cell type
2007-12-21 | GSE7891 | GEO
Project description:The affect of aldosterone on the miRNA landscape in mIMCD-3 cells was determined In the study presented here, the affect of aldosterone was examined on the miRNA content in a murine inner medullary collecting duct cell line
2014-05-31 | E-GEOD-47901 | biostudies-arrayexpress
Project description:Murine inner medullary collecting duct cells were treated for 1 hour with vehicle (control) or aldosterone. Total RNA was isolated and used as template to generate the eventual cRNA target. The experiment was repeated a total of three times. Six cRNA samples, three control and three treated, were generated and used in a total of six hybridizations. The mineralocorticoid aldosterone is a major regulator of Na+ and acid-base balance and control of blood pressure. Although the long-term effects of aldosterone have been extensively studied, the early aldosterone-responsive genes remain largely unknown. Using DNA array technology, we have characterized changes in gene expression after 1 h of exposure to aldosterone in a mouse inner medullary collecting duct cell line, mIMCD-3. Results from three independent microarray experiments revealed that the expression of many transcripts was affected by aldosterone treatment. Northern blot analysis confirmed the upregulation of four distinct transcripts identified by the microarray analysis, namely, the serum and glucose-regulated kinase sgk, connective tissue growth factor, period homolog, and preproendothelin. Immunoblot analysis for preproendothelin demonstrated increased protein expression. Following the levels of the four transcripts over time showed that each had a unique pattern of expression, suggesting that the cellular response to aldosterone is complex. The results presented here represent a novel list of early aldosterone-responsive transcripts and provide new avenues for elucidating the mechanism of acute aldosterone action in the kidney. Keywords: other
2003-05-22 | GSE434 | GEO
Project description:Murine inner medullary collecting duct cells were treated for 1 hour with vehicle (control) or aldosterone. Total RNA was isolated and used as template to generate the eventual cRNA target. The experiment was repeated a total of three times. Six cRNA samples, three control and three treated, were generated and used in a total of six hybridizations. The mineralocorticoid aldosterone is a major regulator of Na+ and acid-base balance and control of blood pressure. Although the long-term effects of aldosterone have been extensively studied, the early aldosterone-responsive genes remain largely unknown. Using DNA array technology, we have characterized changes in gene expression after 1 h of exposure to aldosterone in a mouse inner medullary collecting duct cell line, mIMCD-3. Results from three independent microarray experiments revealed that the expression of many transcripts was affected by aldosterone treatment. Northern blot analysis confirmed the upregulation of four distinct transcripts identified by the microarray analysis, namely, the serum and glucose-regulated kinase sgk, connective tissue growth factor, period homolog, and preproendothelin. Immunoblot analysis for preproendothelin demonstrated increased protein expression. Following the levels of the four transcripts over time showed that each had a unique pattern of expression, suggesting that the cellular response to aldosterone is complex. The results presented here represent a novel list of early aldosterone-responsive transcripts and provide new avenues for elucidating the mechanism of acute aldosterone action in the kidney.
2007-11-06 | E-GEOD-434 | biostudies-arrayexpress
Project description:Regulation of renal water excretion is dependent on control of the molecular water channel aquaporin‐2 by the peptide hormone vasopressin in renal collecting duct cells. The control is in part due to the regulation of transcription of the Aqp2 gene and other genes by vasopressin. A systems biology‐based approach to understanding transcriptional control in renal collecting duct cells depends on knowledge of what transcription factors and other regulatory proteins are present in the cells' nucleus. The goal of this paper is to report comprehensive proteomic profiling of nuclear proteins in native inner medullary collecting duct (IMCD) cells of the rat. Multi‐dimensional separation procedures and state‐of‐the art protein mass spectrometry allowed the high stringency identification of a total of 5105 proteins in nuclear pellet (NP) and nuclear extract (NE) fractions of biochemically isolated rat IMCD cells (URL: https://helixweb.nih.gov/ESBL/Database/IMCD_Nucleus/)*. The analysis identified 369 transcription factor proteins out of the 1371 transcription factors coded by the rat genome. The analysis added 1898 proteins to the recognized proteome of rat IMCD cells, now amounting to 7003 unique proteins. Analysis of samples treated with the vasopressin analog dDAVP (1 nM for 30min) or its vehicle revealed 99 proteins in the NP fraction and 88 proteins in the NE fraction with significant changes in spectral counts (Fisher Exact Test, P<0.005). Among those altered by vasopressin were 7 distinct histone proteins all of which showed decreased abundance in the NP fraction, consistent with a possible effect of vasopressin to induce chromatin remodeling.
2018-10-24 | PXD002680 | Pride
Project description:Phosphorylation of the aquaporin-2 (AQP2) water channel at four COOH-terminal serines plays a central role in the regulation of water permeability of the renal collecting duct. The level of phosphorylation at these sites is determined by a balance between phosphorylation by protein kinases and dephosphorylation by phosphatases. The phosphatases that dephosphorylate AQP2 have not been identified. Here, we use large-scale data integration techniques to identify serine-threonine phosphatases likely to interact with AQP2 in renal collecting duct principal cells. As a first step, we have created a comprehensive list of 38 S/T phosphatase catalytic subunits present in the mammalian genome. Then we used Bayes’ theorem to integrate available information from large-scale data sets from proteomic and transcriptomic studies in order to rank the known S/T phosphatases with regard to the likelihood that they interact with AQP2 in renal collecting duct cells. To broaden the analysis, we have generated new proteomic data (LC-MS/MS) identifying 4538 distinct proteins including 22 S/T phosphatases in cytoplasmic fractions from native inner medullary collecting duct cells from rats. The official gene symbols corresponding to the top-ranked phosphatases (common names in parentheses) were: Ppp1cb (PP1-beta), Ppm1g (PP2C), Ppp1ca (PP1-alpha), Ppp3ca (PP2-B or calcineurin), Ppp2ca (PP2A-alpha), Ppp1cc (PP1-gamma), Ppp2cb (PP2A-beta), Ppp6c (PP6C) and Ppp5c (PP5). This ranking correlates well with results of prior reductionist studies of ion and water channels in renal collecting duct cells.
2018-10-26 | PXD005488 | Pride
Project description:We would like to know the gene expression pattern in absence of transcription factor GATA2 in adult renal collecting duct We used Gata2 flox::Pax8-rtTA::Tet-Cre to make a doxycycline induced Gata2 renal tubule cell specific knockout mice We performed microarray analyses using DBA-lectin and magnetic beads purifed collecting duct cells from WT (n=3) or Gata2 CKO mice (n=3) at 4-weeks after doxycycline induction
2013-11-19 | E-GEOD-52448 | biostudies-arrayexpress
Project description:We applied RNAseq analysis on microdissected outer medullary collecting duct renal segments from mice fed a normal (0.3% Na+) or high Na+ (3%) diet for 3 days. Among 11688 genes sequenced, 293 have an expression significantly modified (p<0.05), 78 with a -1<log2FC>1. In addition to expected results like the decreased expression of genes participating at the Na+ reabsorption (Scnn1a, SGK1), we observed the stimulation of ATP12a gene that participate at a recently described Na+ secretion pathway.
2024-01-01 | GSE244598 | GEO