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Gene expression profiling of osterix-derived cells from control and PTH1R knockout mice in osteoprogenitors

ABSTRACT: Gene expression profiling of osterix-derived cells from control and PTH1R knockout mice in osteoprogenitors

PROVIDER: PRJNA771409 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR16347656_1.fastq.gz	Fastqsanger.gz
	SRR16347656_2.fastq.gz	Fastqsanger.gz
	SRR16347657_1.fastq.gz	Fastqsanger.gz
	SRR16347657_2.fastq.gz	Fastqsanger.gz
	SRR16347658_1.fastq.gz	Fastqsanger.gz

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Project description:The proximal tubule is the sensing site of sodium and phosphorus and the main place for the synthesis and metabolism of 1,25 (OH)2D3. Sodium may share the sensing mechanism with phosphate in proximal tubule epithelial cells; whether sodium cooperates with phosphate or plays an independent role in the regulation of phosphorus homeostasis remains unknown. In this in vitro study, we investigated the effects of high sodium on the synthesis and function of active vitamin D and local phosphorus regulation in proximal tubular epithelial cells. Human proximal tubule epithelial (HK-2) cells were treated with different concentrations of sodium/phosphorus. The expression of 1α-OHase (Cyp27b1) and 24-OHase (Cyp24a1) was determined by RT-PCR and Western Blot. Liquid chromatography/mass spectrometry (LC/MS) and enzyme-linked immunosorbent assay (ELISA) were used to detect the levels of 1,25 (OH)2D3. Intracellular Ca2+ ([Ca2+]i) was detected with the Ca2+ indicator dye Fura-4. Chromatin samples were immunoprecipitated with antibodies against parathyroid receptor 1 (PTH1R) and Klotho. High sodium decreased the expression of 1,25 (OH)2D3 by reducing 1α-OHase and 24-OHase in HK-2 cells, reduced the expression of PTH1R and Klotho, and increased the intracellular calcium concentration. These effects were reversed by sodium phosphorus transporter inhibitor (PFA), sodium hydrogen transporter inhibitor (Caliporide), and a chelator of the extracellular calcium, whereas enhanced by ouabain. Vitamin D receptor agonists significantly increased the recruitment of VDR to the Vitamin D response element (VDRE) of PTH1R and Klotho promoter, thus increasing the expression of PTH1R and Klotho. High extracellular sodium can decrease the synthesis of active vitamin D in the proximal tubules, affect the gene regulation of 1,25 (OH)2D3/VDR, and significantly reduce the expression of PTH1R and Klotho. The data reveal the influence of a high-sodium diet on mineral metabolism and the core role of vitamin D in kidney mineral metabolism.

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Gene expression profiling of osterix-derived cells from control and PTH1R knockout mice in osteoprogenitors

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