Source:http://linkedlifedata.com/resource/pubmed/id/16765983
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
3
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pubmed:dateCreated |
2006-10-18
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pubmed:abstractText |
Despite its multifunctional role in cardiac myocyte function, little is known about dynamic changes in activation state of calmodulin (CaM). Thus, the purpose of this study was to develop a tool to measure Ca bound CaM (Ca-CaM) levels in intact cardiac myocytes. For dynamic measurements of Ca-CaM, we generated an adenoviral vector which expresses a cyan and a yellow fluorescent protein linked by a modified version of the Ca-CaM binding domain of avian smooth muscle myosin light chain kinase. Adult rabbit cardiac myocytes were infected with the Ca-CaM sensing probe or simultaneously infected with viruses containing CaM and the Ca-CaM sensing probe for 24-48 h. Myocytes were then field stimulated (1 Hz) and excited at 440 nm with emitted fluorescence measured at 485 and 535 nm. Changes in [Ca-CaM] are expressed as the ratio of 485 nm/535 nm. Small beat-to-beat changes of [Ca-CaM] were detected, but only when CaM was co-expressed with the sensor. However, upon beta-adrenergic stimulation with isoproterenol, there was an increase in the amplitude of the signals during each beat (parallel to the shortening, which is an indirect measure of [Ca]i) and also a rise in the diastolic [Ca-CaM]. Total [CaM] measured by both competitive ELISA and semi-quantitative Western blots was 5-6 microM in isolated adult ventricular myocytes. Our results indicate that there are dynamic changes in free Ca-CaM levels (a phasic component tracking [Ca]i) as well as system memory that integrates the [Ca]i signals (a tonic component).
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pubmed:grant | |
pubmed:commentsCorrections | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Sep
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pubmed:issn |
0022-2828
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
41
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
451-8
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pubmed:dateRevised |
2007-12-3
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pubmed:meshHeading |
pubmed-meshheading:16765983-Adult,
pubmed-meshheading:16765983-Animals,
pubmed-meshheading:16765983-Biosensing Techniques,
pubmed-meshheading:16765983-Blotting, Western,
pubmed-meshheading:16765983-Calcium,
pubmed-meshheading:16765983-Calmodulin,
pubmed-meshheading:16765983-Cell Line,
pubmed-meshheading:16765983-Humans,
pubmed-meshheading:16765983-Models, Biological,
pubmed-meshheading:16765983-Muscle Cells,
pubmed-meshheading:16765983-Myocardial Contraction,
pubmed-meshheading:16765983-Myocytes, Cardiac,
pubmed-meshheading:16765983-Rabbits,
pubmed-meshheading:16765983-Signal Transduction,
pubmed-meshheading:16765983-Time Factors
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pubmed:year |
2006
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pubmed:articleTitle |
Dynamic changes in free Ca-calmodulin levels in adult cardiac myocytes.
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pubmed:affiliation |
Department of Physiology, Stritch School of Medicine Loyola University, Chicago, 2160 South First Avenue, Maywood, IL 60153, USA.
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pubmed:publicationType |
Journal Article,
Research Support, N.I.H., Extramural
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