TY - JOUR
T1 - Human calcium-sensing receptor can be suppressed by antisense sequences
AU - Maldonado-Pérez, David
AU - Breitwieser, Gerda E.
AU - Gama, Lucio
AU - Elliott, Austin C.
AU - Ward, Donald T.
AU - Riccardi, Daniela
N1 - Funding Information:
This work was supported by the Consejo Nacional de Ciencia y Tecnologı́a (CONACYT) of Mexico and The Overseas Research Students Awards Scheme (2001027016) (studentship to D.M-P). D.T.W. is an NKRF Career development Fellow (TF6/2002). L.G. and G.E.B. were supported by NIH RO1 GM 58578. We thank Dr. Dolores Shoback for the gift of the CaR antibody.
PY - 2003/11/21
Y1 - 2003/11/21
N2 - We have evaluated the ability of an antisense cDNA sequence, directed to the amino-terminus of the human calcium-sensing receptor (CaR), to reduce the expression and function of an EGFP-tagged CaR (CaR-EGFP) in HEK293 cells. Confocal microscopy and Western blot analysis showed a significant and selective reduction of the expression of CaR-EGFP by the antisense construct. Measurements of changes in intracellular calcium induced by CaR agonists showed that CaR-EGFP function was significantly reduced by the antisense sequence, as was agonist-evoked phosphorylation of extracellular signal-regulated protein kinases (ERK1,2). A sense construct directed to the same region of the receptor had no effect, confirming the specificity of the antisense construct. Our results indicate that a CaR antisense cDNA reduces both the expression and function of the receptor. In the absence of strong, specific pharmacological inhibitors of CaR, the antisense approach will be helpful to elucidate contributions of the CaR to cell physiology.
AB - We have evaluated the ability of an antisense cDNA sequence, directed to the amino-terminus of the human calcium-sensing receptor (CaR), to reduce the expression and function of an EGFP-tagged CaR (CaR-EGFP) in HEK293 cells. Confocal microscopy and Western blot analysis showed a significant and selective reduction of the expression of CaR-EGFP by the antisense construct. Measurements of changes in intracellular calcium induced by CaR agonists showed that CaR-EGFP function was significantly reduced by the antisense sequence, as was agonist-evoked phosphorylation of extracellular signal-regulated protein kinases (ERK1,2). A sense construct directed to the same region of the receptor had no effect, confirming the specificity of the antisense construct. Our results indicate that a CaR antisense cDNA reduces both the expression and function of the receptor. In the absence of strong, specific pharmacological inhibitors of CaR, the antisense approach will be helpful to elucidate contributions of the CaR to cell physiology.
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U2 - 10.1016/j.bbrc.2003.10.041
DO - 10.1016/j.bbrc.2003.10.041
M3 - Article
C2 - 14623314
AN - SCOPUS:0242577586
SN - 0006-291X
VL - 311
SP - 610
EP - 617
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -