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CA²⁺-activated potassium channels in smooth muscle cell from cerebral artery of adult rat Wang, Yihong
Abstract
Patch clamp methods were used to study the biophysical properties of Ca ²+ -activated potassium (KCa) channels in cerebrovascular smooth muscle cells (CVSMCs) derived from the cerebral arteries of adult rats. Procedures were developed for the enzymatic dissociation of CVSMCs from cerebral arteries. Dissociated cells were maintained at 4°C or cultured at 37°C for 1-3 days prior to use. CVSMCs were identified using a monoclonal antibody specific for smooth muscle a -actin. The calcium-sensitive fluorescent probe fura-2 was employed to measure tht resting and serotonin (5-HT) stimulated levels of free intracellular calcium, [Ca ²+ ] in these cells. A resting [Ca ²+ ]i level of 41 ± 5.6 nM was obtained from a total of 110 CVSMCs in culture. Serotonin (5-HT, 10 nM to 100μ M) induced a transient increase of [Ca ²+Ji above resting levels. The effects of three known blockers of voltage-dependent Ca ²+ channels, nifedipine, La³ + and Co²+ were tested by coapplication with 5-HT. It was found the neither 10 mM LaC13 nor 10 mM C0C12 reduced the rise in [Ca ²+ ft evoked by application of 1 μ M 5-HT. Nifedipine (10 μ M) also failed to significantly reduce the rise in [Ca ²+ ]i activated by 1 μ M5-HT. The effects of the partially selective 5-HT₂ receptor antagonist, ketanserin (5 nM) reversibly attenuated the 5-HT response in all cells tested. The effect of 5-HT on [Ca ²+ Ji was clearly dose-dependent and the concentration of 5- HT producing a half-maximal increase in [Ca ²+ ]i was found to be 10 nM.[more abstract]
Item Metadata
Title |
CA²⁺-activated potassium channels in smooth muscle cell from cerebral artery of adult rat
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Creator | |
Publisher |
University of British Columbia
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Date Issued |
1992
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Description |
Patch clamp methods were used to study the biophysical properties of
Ca ²+ -activated potassium (KCa) channels in cerebrovascular smooth muscle cells
(CVSMCs) derived from the cerebral arteries of adult rats. Procedures were
developed for the enzymatic dissociation of CVSMCs from cerebral arteries.
Dissociated cells were maintained at 4°C or cultured at 37°C for 1-3 days prior to
use. CVSMCs were identified using a monoclonal antibody specific for smooth
muscle a -actin. The calcium-sensitive fluorescent probe fura-2 was employed to
measure tht resting and serotonin (5-HT) stimulated levels of free intracellular
calcium, [Ca ²+ ] in these cells.
A resting [Ca ²+ ]i level of 41 ± 5.6 nM was obtained from a total of 110
CVSMCs in culture. Serotonin (5-HT, 10 nM to 100μ M) induced a transient
increase of [Ca ²+Ji above resting levels. The effects of three known blockers of
voltage-dependent Ca ²+ channels, nifedipine, La³ + and Co²+ were tested by
coapplication with 5-HT. It was found the neither 10 mM LaC13 nor 10 mM
C0C12 reduced the rise in [Ca ²+ ft evoked by application of 1 μ M 5-HT.
Nifedipine (10 μ M) also failed to significantly reduce the rise in [Ca ²+ ]i activated
by 1 μ M5-HT. The effects of the partially selective 5-HT₂ receptor antagonist,
ketanserin (5 nM) reversibly attenuated the 5-HT response in all cells tested. The
effect of 5-HT on [Ca ²+ Ji was clearly dose-dependent and the concentration of 5-
HT producing a half-maximal increase in [Ca ²+ ]i was found to be 10 nM.[more abstract]
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Extent |
4532454 bytes
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Genre | |
Type | |
File Format |
application/pdf
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Language |
eng
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Date Available |
2008-12-17
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Provider |
Vancouver : University of British Columbia Library
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Rights |
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.
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DOI |
10.14288/1.0086586
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URI | |
Degree | |
Program | |
Affiliation | |
Degree Grantor |
University of British Columbia
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Graduation Date |
1992-05
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Campus | |
Scholarly Level |
Graduate
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Aggregated Source Repository |
DSpace
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Item Media
Item Citations and Data
Rights
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.