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Dissecting hypertonicity- and NFAT5-dependent gene expression programs in mpkCCD cells
Kristina Engel1, Dmitry Chernyakov1, Moritz Pernecker2
1Department of Medicine, Hematology and Oncology, Martin Luther University Halle-Wittenberg, Halle (Saale), Germany.
Abstract:
The corticomedullary osmotic gradient between renal cortex and medulla induces a specific spatial gene expression pattern. The factors controlling these differences have not been fully addressed. A hypertonic environment leads to the activation of nuclear factor of activated T-cells 5 (NFAT5), which regulates the expression of osmoprotective genes. While NFAT5 function under hypertonic conditions has been extensively studied, its contribution to basal gene regulation remains unclear. We used murine principal kidney cortical collecting duct (mpkCCD) cells, induced functional deletion of NFAT5, and performed gene expression profiling to identify genes that are differentially expressed under isotonic and hypertonic cell culture conditions. Hypertonic stress induced extensive transcriptional changes in control cells, which were markedly altered in NFAT5-deficient cells. Furthermore, a comparison of the mpkCCD transcriptomes with gene expression profiles from the renal cortex and inner medulla of control and principal cell-specific NFAT5 knockout mice revealed a partial overlap in hypertonicity-associated and NFAT5-dependent gene expression patterns. In both conditions, the expression of known NFAT5 target genes, like Aqp2 and Ranbp3l, was downregulated. These findings support the use of mpkCCD cells as a complementary model for studying NFAT5-associated gene regulation under controlled in vitro conditions.
Insights
Nuclear factor of activated T-cells 5 (NFAT5) regulates gene expression in kidney cells. This study reveals NFAT5
Area of Science:
- Renal physiology and molecular biology
- Kidney gene expression regulation
Background:
- The corticomedullary osmotic gradient influences kidney gene expression.
- Nuclear factor of activated T-cells 5 (NFAT5) is activated by hypertonicity and regulates osmoprotective genes.
- NFAT5's role in basal gene regulation is not fully understood.
Purpose of the Study:
- To investigate the role of NFAT5 in gene regulation under both isotonic and hypertonic conditions.
- To identify genes regulated by NFAT5 in kidney cells.
- To compare in vitro findings with in vivo mouse models.
Main Methods:
- Utilized murine principal kidney cortical collecting duct (mpkCCD) cells.
- Induced functional deletion of NFAT5 in mpkCCD cells.
- Performed gene expression profiling under isotonic and hypertonic conditions.
- Compared transcriptomes with gene expression data from NFAT5 knockout mice.
Main Results:
- Hypertonic stress caused significant transcriptional changes in control cells, altered in NFAT5-deficient cells.
- A partial overlap was observed between hypertonicity-associated, NFAT5-dependent gene expression in vitro and in vivo.
- Known NFAT5 target genes, such as Aqp2 and Ranbp3l, were downregulated in NFAT5-deficient conditions.
Conclusions:
- NFAT5 plays a role in regulating gene expression under both basal and hypertonic conditions.
- mpkCCD cells serve as a valuable complementary model for studying NFAT5-mediated gene regulation in vitro.
- Findings provide insights into the spatial gene expression patterns in the renal medulla.
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