Details

Autor(en) / Beteiligte

• Shirokova, Natalia
• Shirokov, Roman
• Rossi, Daniela
• González, Adom
• Kirsch, Wolfgang G.
• García, Jesús
• Sorrentino, Vincenzo
• Ríos, Eduardo

Titel

Spatially segregated control of Ca2+ release in developing skeletal muscle of mice

Ist Teil von

• The Journal of physiology, 1999-12, Vol.521 (2), p.483-495

Ort / Verlag

Oxford, UK: The Physiological Society

Erscheinungsjahr

1999

Link zum Volltext

Quelle

Wiley Online Library - AutoHoldings Journals

Beschreibungen/Notizen

• Confocal laser scanning microscopy was used to monitor Ca 2+ signals in primary-cultured myotubes, prepared from forelimbs of wild-type or ryanodine receptor type 3 (RyR3) knockout mice. Myotubes loaded with the acetoxymethyl ester (AM) form of fluo-3 were imaged at rest or under whole-cell patch clamp. Discrete Ca 2+ release events were detected in intact wild-type and RyR3-knockout myotubes. They showed almost no difference in amplitude and width, but were substantially different in duration. In wild-type myotubes (660 events, 57 cells) the amplitude was 1.27 (0.85, 1.97) (median (25%, 75%)) units of resting fluorescence, the full width at half-magnitude (FWHM) was 1.4 (0.9, 2.3) μm, and the full duration at half-magnitude (FDHM) was 25.3 (9.6, 51.7) ms. In RyR3-knockout myotubes (655 events, 83 cells) the amplitude was 1.30 (0.84, 2.08), FWHM was 1.63 (1.02, 2.66) μm, and FDHM was 43.6 (23.6, 76.9) ms. Depolarization under voltage clamp of both wild-type and RyR3-knockout myotubes produced substantial Ca 2+ release devoid of discrete Ca 2+ events. Discrete events were still present but occurred without correlation with the applied pulse, largely at locations where the pulse did not elicit release. The local correspondence between voltage control and absence of discrete events implies that the functional interaction with voltage sensors suppresses the mechanism that activates discrete events. Because it applies whether RyR3 is present or not, it is this exclusion by voltage of other control mechanisms, rather than isoform composition, that primarily determines the absence of discrete Ca 2+ events in adult mammalian muscle.