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Cyclic ADP-ribose does not affect cardiac or skeletal muscle ryanodine receptors
B R Fruen1, J R Mickelson, N H Shomer
1Department of Veterinary PathoBiology, University of Minnesota, St. Paul 55108.
Abstract:
The cardiac muscle isoform of the ryanodine receptor/Ca2+ release channel (RYR) has been proposed to be an important target of cyclic ADP-ribose (cADPR) action in mammalian cells. However, we now demonstrate that neither cADPR (0.1-5 microM), nor the related metabolites beta-NAD+ (0.1-30 mM) and ADP-ribose (0.1-5 microM), affected cardiac RYR activity as determined by [3H]ryanodine binding to cardiac sarcoplasmic reticulum (SR) vesicles. Similarly, cADPR (1 microM) failed to activate single cardiac RYR channels in planar lipid bilayers. Skeletal muscle SR [3H]ryanodine binding was also unaffected by cADPR (up to 30 microM). These results argue against a direct role for the well-characterized RYRs of cardiac or skeletal muscle in mediating cADPR-activated Ca2+ release.
Insights
Cyclic ADP-ribose (cADPR) does not directly affect cardiac or skeletal ryanodine receptors (RYR). This study found no evidence that cADPR influences RYR activity in mammalian cardiac and skeletal muscle sarcoplasmic reticulum.
Area of Science:
- Molecular biology
- Cellular physiology
- Biochemistry
Background:
- The cardiac ryanodine receptor (RYR) is a calcium (Ca2+) release channel in mammalian cells.
- Cyclic ADP-ribose (cADPR) has been proposed to target cardiac RYR channels.
- Understanding RYR regulation is crucial for cardiac function.
Purpose of the Study:
- To investigate the direct effect of cADPR on cardiac and skeletal muscle ryanodine receptor (RYR) activity.
- To determine if cADPR modulates Ca2+ release through RYRs in mammalian muscle tissues.
Main Methods:
- [3H]ryanodine binding assays on cardiac and skeletal muscle sarcoplasmic reticulum (SR) vesicles.
- Single channel recordings of cardiac RYR activity in planar lipid bilayers.
- Application of varying concentrations of cADPR, beta-NAD+, and ADP-ribose.
Main Results:
- Neither cADPR, beta-NAD+, nor ADP-ribose affected [3H]ryanodine binding to cardiac SR vesicles.
- cADPR did not activate single cardiac RYR channels in lipid bilayers.
- cADPR did not affect skeletal muscle SR [3H]ryanodine binding.
Conclusions:
- The findings argue against a direct role for cardiac and skeletal muscle RYRs in mediating cADPR-activated Ca2+ release.
- This study challenges the proposed mechanism of cADPR action on well-characterized RYRs.
- Further research is needed to elucidate the precise role of cADPR in cellular Ca2+ signaling.