CRAYFISH SKELETAL MUSCLE REQUIRES BOTH INFLUX OF EXTERNAL Ca2+ AND Ca2+ RELEASE FROM INTERNAL STORES FOR CONTRACTION

1993 ◽  
Vol 181 (1) ◽  
pp. 95-105 ◽  
Author(s):  
H. Ushio ◽  
S. Watabe ◽  
M. Iino

The isometric tension and membrane potential of single skeletal muscle fibres from the flexor muscle of the carpopodite in the meropodite of crayfish Procambarus clarkii (Girard) were studied to determine whether crayfish muscle contraction requires Ca2+ release from the sarcoplasmic reticulum. Contraction elicited by brief extracellular electrical stimulation was reduced by the removal of Ca2+ or by the addition of 25 micromolar nicardipine in crayfish Ringer's solution. Addition of 30 micromolar ryanodine with 1 mmol l-1 caffeine induced a transient contracture, the peak tension of which was 10–30 % of that of the high-K+ induced contracture and which declined to the pretreatment level in 20–60 min. After ryanodine-caffeine treatment, 30 mmol l-1 caffeine failed to induce contraction, suggesting that intracellular Ca2+ stores had been exhausted by the treatment. Extracellular electrical stimulation also failed to induce contraction after ryanodine-caffeine treatment, although the resting potential was not changed. These results suggest that Ca2+ release from the sarcoplasmic reticulum, together with Ca2+ influx via nicardipine-sensitive Ca2+ channels, is essential to the contraction of crayfish leg muscle fibres after a brief membrane depolarization.

2004 ◽  
Vol 379 (2) ◽  
pp. 505-512 ◽  
Author(s):  
Alessandra NORI ◽  
Elena BORTOLOSO ◽  
Federica FRASSON ◽  
Giorgia VALLE ◽  
Pompeo VOLPE

CS (calsequestrin) is an acidic glycoprotein of the SR (sarcoplasmic reticulum) lumen and plays a crucial role in the storage of Ca2+ and in excitation–contraction coupling of skeletal muscles. CS is synthesized in the ER (endoplasmic reticulum) and is targeted to the TC (terminal cisternae) of SR via mechanisms still largely unknown, but probably involving vesicle transport through the Golgi complex. In the present study, two mutant forms of Sar1 and ARF1 (ADP-ribosylation factor 1) were used to disrupt cargo exit from ER-exit sites and intra-Golgi trafficking in skeletal-muscle fibres respectively. Co-expression of Sar1-H79G (His79→Gly) and recombinant, epitope-tagged CS, CSHA1 (where HA1 stands for nine-amino-acid epitope of the viral haemagglutinin 1), barred segregation of CSHA1 to TC. On the other hand, expression of ARF1-N126I altered the subcellular localization of GM130, a cis-medial Golgi protein in skeletal-muscle fibres and myotubes, without interfering with CSHA1 targeting to either TC or developing SR. Thus active budding from ER-exit sites appears to be involved in CS targeting and routing, but these processes are insensitive to modification of intracellular vesicle trafficking and Golgi complex disruption caused by the mutant ARF1-N126I. It also appears that CS routing from ER to SR does not involve classical secretory pathways through ER–Golgi intermediate compartments, cis-medial Golgi and trans-Golgi network.


2010 ◽  
Vol 459 (5) ◽  
pp. 725-735 ◽  
Author(s):  
Ramón Jiménez-Moreno ◽  
Zhong-Ming Wang ◽  
María Laura Messi ◽  
Osvaldo Delbono

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