Voltage-dependent calcium channels of dog basilar artery

Elena Nikitina, Zhen Du Zhang, Ayako Kawashima, Babak S. Jahromi, Vitali A. Bouryi, Masataka Takahashi, An Xie, R. Loch Macdonald*

*Corresponding author for this work

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Electrophysiological and molecular characteristics of voltage-dependent calcium (Ca2+) channels were studied using whole-cell patch clamp, polymerase chain reaction and Western blotting in smooth muscle cells freshly isolated from dog basilar artery. Inward currents evoked by depolarizing steps from a holding potential of -50 or -90 mV in 10 m m barium consisted of low- (LVA) and high-voltage activated (HVA) components. LVA current comprised more than half of total current in 24 (12%) of 203 cells and less than 10% of total current in 52 (26%) cells. The remaining cells (127 cells, 62%) had LVA currents between one tenth and one half of total current. LVA current was rapidly inactivating, slowly deactivating, inhibited by high doses of nimodipine and mibefradil (> 0.3 μm), not affected by ω-agatoxin GVIA (γ100 nm), ω-conotoxin IVA (1 μm) or SNX-482 (200 nm) and probably carried by T-type Ca2+ channels based on the presence of messenger ribonucleic acid (mRNA) and protein for Cav3.1 and Cav3.3 α 1 subunits of these channels. LVA currents exhibited window current with a maximum of 13% of the LVA current at -37.4 mV. HVA current was slowly inactivating and rapidly deactivating. It was inhibited by nimodipine (IC50 = 0.018 μm), mibefradil (IC50 = 0.39 μm) and ω-conotoxin IV (1 μm). Smooth muscle cells also contained mRNA and protein for L- (Cav1.2 and Cav1.3), N- (Cav2.2) and T-type (Cav3.1 and Cav3.3) α1 Ca2+ channel subunits. Confocal microscopy showed Cav1.2 and Cav1.3 (L-type), Cav2.2 (N-type) and Cav3.1 and Cav3.3 (T-type) protein in smooth muscle cells. Relaxation of intact arteries under isometric tension in vitro to nimodipine (1 μM) and mibefradil (1 μM) but not to ω-agatoxin GVIA (100 nm), ω-conotoxin IVA (1 μM) or SNX-482 (1 μM) confirmed the functional significance of L- and T-type voltage-dependent Ca2+ channel subtypes but not N-type. These results show that dog basilar artery smooth muscle cells express functional voltage-dependent Ca2+ channels of multiple types.

Original languageEnglish (US)
Pages (from-to)523-541
Number of pages19
JournalJournal of Physiology
Volume580
Issue number2
DOIs
StatePublished - Apr 15 2007

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Basilar Artery
Calcium Channels
Mibefradil
Nimodipine
Smooth Muscle Myocytes
Dogs
Agatoxins
Conotoxins
Inhibitory Concentration 50
RNA
Proteins
Barium
Confocal Microscopy
Arteries
Western Blotting
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Physiology

Cite this

Nikitina, E., Zhang, Z. D., Kawashima, A., Jahromi, B. S., Bouryi, V. A., Takahashi, M., ... Macdonald, R. L. (2007). Voltage-dependent calcium channels of dog basilar artery. Journal of Physiology, 580(2), 523-541. https://doi.org/10.1113/jphysiol.2006.126128
Nikitina, Elena ; Zhang, Zhen Du ; Kawashima, Ayako ; Jahromi, Babak S. ; Bouryi, Vitali A. ; Takahashi, Masataka ; Xie, An ; Macdonald, R. Loch. / Voltage-dependent calcium channels of dog basilar artery. In: Journal of Physiology. 2007 ; Vol. 580, No. 2. pp. 523-541.
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abstract = "Electrophysiological and molecular characteristics of voltage-dependent calcium (Ca2+) channels were studied using whole-cell patch clamp, polymerase chain reaction and Western blotting in smooth muscle cells freshly isolated from dog basilar artery. Inward currents evoked by depolarizing steps from a holding potential of -50 or -90 mV in 10 m m barium consisted of low- (LVA) and high-voltage activated (HVA) components. LVA current comprised more than half of total current in 24 (12{\%}) of 203 cells and less than 10{\%} of total current in 52 (26{\%}) cells. The remaining cells (127 cells, 62{\%}) had LVA currents between one tenth and one half of total current. LVA current was rapidly inactivating, slowly deactivating, inhibited by high doses of nimodipine and mibefradil (> 0.3 μm), not affected by ω-agatoxin GVIA (γ100 nm), ω-conotoxin IVA (1 μm) or SNX-482 (200 nm) and probably carried by T-type Ca2+ channels based on the presence of messenger ribonucleic acid (mRNA) and protein for Cav3.1 and Cav3.3 α 1 subunits of these channels. LVA currents exhibited window current with a maximum of 13{\%} of the LVA current at -37.4 mV. HVA current was slowly inactivating and rapidly deactivating. It was inhibited by nimodipine (IC50 = 0.018 μm), mibefradil (IC50 = 0.39 μm) and ω-conotoxin IV (1 μm). Smooth muscle cells also contained mRNA and protein for L- (Cav1.2 and Cav1.3), N- (Cav2.2) and T-type (Cav3.1 and Cav3.3) α1 Ca2+ channel subunits. Confocal microscopy showed Cav1.2 and Cav1.3 (L-type), Cav2.2 (N-type) and Cav3.1 and Cav3.3 (T-type) protein in smooth muscle cells. Relaxation of intact arteries under isometric tension in vitro to nimodipine (1 μM) and mibefradil (1 μM) but not to ω-agatoxin GVIA (100 nm), ω-conotoxin IVA (1 μM) or SNX-482 (1 μM) confirmed the functional significance of L- and T-type voltage-dependent Ca2+ channel subtypes but not N-type. These results show that dog basilar artery smooth muscle cells express functional voltage-dependent Ca2+ channels of multiple types.",
author = "Elena Nikitina and Zhang, {Zhen Du} and Ayako Kawashima and Jahromi, {Babak S.} and Bouryi, {Vitali A.} and Masataka Takahashi and An Xie and Macdonald, {R. Loch}",
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Nikitina, E, Zhang, ZD, Kawashima, A, Jahromi, BS, Bouryi, VA, Takahashi, M, Xie, A & Macdonald, RL 2007, 'Voltage-dependent calcium channels of dog basilar artery', Journal of Physiology, vol. 580, no. 2, pp. 523-541. https://doi.org/10.1113/jphysiol.2006.126128

Voltage-dependent calcium channels of dog basilar artery. / Nikitina, Elena; Zhang, Zhen Du; Kawashima, Ayako; Jahromi, Babak S.; Bouryi, Vitali A.; Takahashi, Masataka; Xie, An; Macdonald, R. Loch.

In: Journal of Physiology, Vol. 580, No. 2, 15.04.2007, p. 523-541.

Research output: Contribution to journalArticle

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AU - Nikitina, Elena

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AU - Kawashima, Ayako

AU - Jahromi, Babak S.

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AU - Takahashi, Masataka

AU - Xie, An

AU - Macdonald, R. Loch

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N2 - Electrophysiological and molecular characteristics of voltage-dependent calcium (Ca2+) channels were studied using whole-cell patch clamp, polymerase chain reaction and Western blotting in smooth muscle cells freshly isolated from dog basilar artery. Inward currents evoked by depolarizing steps from a holding potential of -50 or -90 mV in 10 m m barium consisted of low- (LVA) and high-voltage activated (HVA) components. LVA current comprised more than half of total current in 24 (12%) of 203 cells and less than 10% of total current in 52 (26%) cells. The remaining cells (127 cells, 62%) had LVA currents between one tenth and one half of total current. LVA current was rapidly inactivating, slowly deactivating, inhibited by high doses of nimodipine and mibefradil (> 0.3 μm), not affected by ω-agatoxin GVIA (γ100 nm), ω-conotoxin IVA (1 μm) or SNX-482 (200 nm) and probably carried by T-type Ca2+ channels based on the presence of messenger ribonucleic acid (mRNA) and protein for Cav3.1 and Cav3.3 α 1 subunits of these channels. LVA currents exhibited window current with a maximum of 13% of the LVA current at -37.4 mV. HVA current was slowly inactivating and rapidly deactivating. It was inhibited by nimodipine (IC50 = 0.018 μm), mibefradil (IC50 = 0.39 μm) and ω-conotoxin IV (1 μm). Smooth muscle cells also contained mRNA and protein for L- (Cav1.2 and Cav1.3), N- (Cav2.2) and T-type (Cav3.1 and Cav3.3) α1 Ca2+ channel subunits. Confocal microscopy showed Cav1.2 and Cav1.3 (L-type), Cav2.2 (N-type) and Cav3.1 and Cav3.3 (T-type) protein in smooth muscle cells. Relaxation of intact arteries under isometric tension in vitro to nimodipine (1 μM) and mibefradil (1 μM) but not to ω-agatoxin GVIA (100 nm), ω-conotoxin IVA (1 μM) or SNX-482 (1 μM) confirmed the functional significance of L- and T-type voltage-dependent Ca2+ channel subtypes but not N-type. These results show that dog basilar artery smooth muscle cells express functional voltage-dependent Ca2+ channels of multiple types.

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Nikitina E, Zhang ZD, Kawashima A, Jahromi BS, Bouryi VA, Takahashi M et al. Voltage-dependent calcium channels of dog basilar artery. Journal of Physiology. 2007 Apr 15;580(2):523-541. https://doi.org/10.1113/jphysiol.2006.126128