Antisense oligonucleotide against the Ca channel β subunit decreases L-type Ca current in rat ventricular myocytes

Rob N. Leach; Fabien Brette; Clive H. Orchard

doi:10.1016/j.bbrc.2006.11.099

Antisense oligonucleotide against the Ca channel β subunit decreases L-type Ca current in rat ventricular myocytes

Rob N. Leach, Fabien Brette, Clive H. Orchard^*

^*Corresponding author for this work

Research output: Contribution to journal › Article (Academic Journal) › peer-review

2 Citations (Scopus)

Abstract

The role of endogenous β subunits of the L-type Ca channel in native cardiac ventricular myocytes is unclear. We have therefore investigated the effect of inhibiting β subunit expression in rat myocytes, by culturing isolated myocytes for 24 h with either antisense oligonucleotide against the β subunit or with scrambled oligonucleotide (control). α₁ subunit expression and distribution were then determined by immunolabeling, and L-type Ca current measured using the whole cell patch-clamp technique. Cells treated with antisense showed increased perinuclear staining for α₁, decreased Ca current amplitude and a small rightward shift of the activation curve and the I-V relation, with no significant effect on inactivation. These data suggest that endogenous β subunits in native cardiac myocytes help to traffic the α₁ subunit to the cell membrane and thus play a major role in determining Ca current amplitude.

Original language	English
Pages (from-to)	794-798
Number of pages	5
Journal	Biochemical and Biophysical Research Communications
Volume	352
Issue number	3
DOIs	https://doi.org/10.1016/j.bbrc.2006.11.099
Publication status	Published - 19 Jan 2007

Keywords

β Subunit
Calcium current
Cardiac myocytes
L-type Ca channel

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title = "Antisense oligonucleotide against the Ca channel β subunit decreases L-type Ca current in rat ventricular myocytes",

abstract = "The role of endogenous β subunits of the L-type Ca channel in native cardiac ventricular myocytes is unclear. We have therefore investigated the effect of inhibiting β subunit expression in rat myocytes, by culturing isolated myocytes for 24 h with either antisense oligonucleotide against the β subunit or with scrambled oligonucleotide (control). α1 subunit expression and distribution were then determined by immunolabeling, and L-type Ca current measured using the whole cell patch-clamp technique. Cells treated with antisense showed increased perinuclear staining for α1, decreased Ca current amplitude and a small rightward shift of the activation curve and the I-V relation, with no significant effect on inactivation. These data suggest that endogenous β subunits in native cardiac myocytes help to traffic the α1 subunit to the cell membrane and thus play a major role in determining Ca current amplitude.",

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AU - Leach, Rob N.

AU - Brette, Fabien

AU - Orchard, Clive H.

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N2 - The role of endogenous β subunits of the L-type Ca channel in native cardiac ventricular myocytes is unclear. We have therefore investigated the effect of inhibiting β subunit expression in rat myocytes, by culturing isolated myocytes for 24 h with either antisense oligonucleotide against the β subunit or with scrambled oligonucleotide (control). α1 subunit expression and distribution were then determined by immunolabeling, and L-type Ca current measured using the whole cell patch-clamp technique. Cells treated with antisense showed increased perinuclear staining for α1, decreased Ca current amplitude and a small rightward shift of the activation curve and the I-V relation, with no significant effect on inactivation. These data suggest that endogenous β subunits in native cardiac myocytes help to traffic the α1 subunit to the cell membrane and thus play a major role in determining Ca current amplitude.

AB - The role of endogenous β subunits of the L-type Ca channel in native cardiac ventricular myocytes is unclear. We have therefore investigated the effect of inhibiting β subunit expression in rat myocytes, by culturing isolated myocytes for 24 h with either antisense oligonucleotide against the β subunit or with scrambled oligonucleotide (control). α1 subunit expression and distribution were then determined by immunolabeling, and L-type Ca current measured using the whole cell patch-clamp technique. Cells treated with antisense showed increased perinuclear staining for α1, decreased Ca current amplitude and a small rightward shift of the activation curve and the I-V relation, with no significant effect on inactivation. These data suggest that endogenous β subunits in native cardiac myocytes help to traffic the α1 subunit to the cell membrane and thus play a major role in determining Ca current amplitude.

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KW - Calcium current

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