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Human calcium-sensing receptor can be suppressed by antisense sequences.
David Maldonado-Pérez1, Gerda E Breitwieser, Lucio Gama
1School of Biological Sciences, University of Manchester, Manchester, UK.
Biochemical and Biophysical Research Communications
|November 19, 2003
Summary
An antisense cDNA sequence effectively reduced human calcium-sensing receptor (CaR) expression and function in cells. This approach offers a valuable tool for studying CaR
Area of Science:
- Molecular Biology
- Cell Biology
- Pharmacology
Background:
- The human calcium-sensing receptor (CaR) plays a critical role in calcium homeostasis.
- Specific pharmacological inhibitors for CaR are limited, hindering research into its physiological contributions.
Purpose of the Study:
- To evaluate the efficacy of an antisense cDNA sequence targeting the CaR amino-terminus.
- To assess the impact of the antisense construct on CaR expression and function in HEK293 cells.
Main Methods:
- Utilized an antisense cDNA construct targeting the amino-terminus of the human CaR.
- Employed confocal microscopy and Western blot analysis to assess CaR expression.
- Measured intracellular calcium changes and ERK1,2 phosphorylation in response to CaR agonists.
Main Results:
- The antisense construct significantly and selectively reduced CaR-EGFP expression.
- CaR-EGFP function, including agonist-induced calcium signaling and ERK1,2 phosphorylation, was significantly impaired.
- A sense construct showed no effect, confirming the specificity of the antisense approach.
Conclusions:
- Antisense cDNA targeting CaR effectively reduces both receptor expression and function.
- This antisense strategy provides a valuable tool for investigating the role of CaR in cellular physiology.
- The findings support the utility of antisense technology in the absence of specific pharmacological CaR modulators.