What question did this study set out to answer?

To determine whether the expression of ryanodine receptor type 3 (RyR3) is sufficient to generate discrete elementary Ca2+ release events in skeletal myotubes lacking all endogenous RyR isoforms.

May 1, 2000The Journal of PhysiologyOpen Access

Expression of ryanodine receptor RyR3 produces Ca2+ sparks in dyspedic myotubes

Q: What is the clinical evidence from this study?

Study design: Other. Intervention: Expression of ryanodine receptor RyR3 vs. Non-transduced RyR null myotubes. Primary outcome: Production of Ca2+ sparks.

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Key result

Expression of ryanodine receptor RyR3 is sufficient for the production of Ca2+ sparks in a skeletal muscle system lacking the expression of any other RyR isoform.

Population

'dyspedic' 1B5 myotubes (myotubes which do not express ryanodine receptors)

Comparison

Transduction with virions containing cDNA for… vs Non-transduced RyR null myotubes

Design

Preclinical

Authors

CWChristopher W. WardGeneral Cardiology MSMartin F. SchneiderUniversity of Maryland, Baltimore DCDaniel Castillo-SecillaFujitsu (Japan)

Discussion

Member takes

Implication

RyR3 enables Ca2+ sparks in RyR-null skeletal muscle; extends basic isoform mechanisms but leaves open human disease relevance.

Key Points

To determine whether the expression of ryanodine receptor type 3 (RyR3) is sufficient to generate discrete elementary Ca2+ release events in skeletal myotubes lacking all endogenous RyR isoforms.
Transduced dyspedic 1B5 myotubes (RyR-null) with viral vectors encoding RyR3 cDNA.
Permeabilized myotubes with saponin, introduced the fluorescent indicator fluo-3, and imaged Ca2+ sparks using laser scanning confocal microscopy in line scan (xt) and planar (xy) modes.
Correlated spark locations with RyR3 distribution using post-recording fixation and antibody staining, comparing spark kinetics directly to permeabilized frog skeletal muscle fibers.
Ca2+ sparks were entirely absent in non-transduced RyR-null myotubes but readily appeared following RyR3 transduction, co-localizing with punctate peripheral RyR3 antibody staining.
Permeabilized RyR3-expressing myotubes produced Ca2+ sparks with a mean amplitude (ΔF/F) of 1.20 ± 0.04 and a 10–90% rise time of 6.31 ± 0.12 ms, compared to 0.98 ± 0.01 and 6.11 ± 0.07 ms in frog skeletal muscle fibers.
RyR3-mediated sparks showed approximately 40% larger spatial extent and duration than frog muscle sparks, frequently recurring at identical sites with reproducible amplitudes and spatio-temporal properties.

Structured PICO

Population

'dyspedic' 1B5 myotubes (myotubes which do not express ryanodine receptors)

Intervention

Transduction with virions containing cDNA for RyR type 3

Comparator

Non-transduced RyR null myotubes

Outcome

Production and characteristics of Ca2+ sparks (amplitude, temporal rise times, spatial extent, temporal duration)surrogate

The presence of RyR3 is sufficient for the production of Ca2+ sparks in a skeletal muscle system lacking the expression of any other RyR isoform.

Cite This Study

Ward et al. (2000) studied this question. Expression of ryanodine receptor RyR3 vs. Non-transduced RyR null myotubes was evaluated on Production of Ca2+ sparks. Expression of ryanodine receptor RyR3 is sufficient for the production of Ca2+ sparks in a skeletal muscle system lacking the expression of any other RyR isoform.

synapsesocial.com/papers/6a957f057007bd2ef4e3cbc4 https://doi.org/10.1111/j.1469-7793.2000.t01-2-00091.x

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Expression of ryanodine receptor RyR3 produces Ca2+ sparks in dyspedic myotubes

Discussion

Member takes

Implication

Key Points

Structured PICO

Cite This Study

Expression of ryanodine receptor RyR3 produces Ca2+ sparks in dyspedic myotubes

Discussion

Member takes

Implication

Key Points

Structured PICO

Cite This Study

Also Consider

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