Hormonally-regulated expression of voltage-operated Ca2+ channels in osteocytic (MLO-Y4) cells

Y Gu, MR Preston, J Magnay, AJ El Haj… - Biochemical and …, 2001 - Elsevier
Y Gu, MR Preston, J Magnay, AJ El Haj, SJ Publicover
Biochemical and biophysical research communications, 2001Elsevier
Voltage operated calcium channels (VOCCs) are important in stimulus-response coupling in
osteoblasts. We have investigated the expression of VOCCs in the mouse osteocyte cell
line, MLO-Y4. Using the whole-cell patch clamp technique we were unable to detect any
VOCC currents (n= 436) even in the presence of the L-type VOCC agonist Bay K 8644 (n=
350). Reverse transcription polymerase chain reaction (RT-PCR), using primers to detect
α1C, α1D, and α1G VOCC subunits (all of which are expressed in primary osteoblasts), did …
Voltage operated calcium channels (VOCCs) are important in stimulus-response coupling in osteoblasts. We have investigated the expression of VOCCs in the mouse osteocyte cell line, MLO-Y4. Using the whole-cell patch clamp technique we were unable to detect any VOCC currents (n = 436) even in the presence of the L-type VOCC agonist Bay K 8644 (n = 350). Reverse transcription polymerase chain reaction (RT-PCR), using primers to detect α1C, α1D, and α1G VOCC subunits (all of which are expressed in primary osteoblasts), did not generate detectable products with mRNA from MLO-Y4 cells. However, after treatment with physiological levels of hormones, VOCC α1 subunit mRNAs were detected in MLO-Y4 cells. PTH, 17β-estradiol, and dexamethasone-treatment induced expression of L-type (α1C, α1D) subunit transcripts. ATP-treatment induced expression of T-type (α1G) transcripts. Using whole-cell patch clamp we detected VOCC currents in 5–10% of cells after treatment. Current characteristics (L- or T-type) were consistent with the transcript expressed.
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