Dr. Claflin et al., THE EFFECT OF LENGTH ON THE RELATIONSHIP BETWEEN TENSION AND INTRACELLULAR [CA2-MUSCLE FIBERS(] IN INTACT FROG SKELETAL), Journal of physiology, 508(1), 1998, pp. 179-186
1. The relationship between tension and intracellular calcium concentr
ation ([Ca2+](i)) in intact frog skeletal muscle fibres was determined
at two fibre lengths, corresponding to mean sarcomere lengths (SL) of
2.2 and 2.9 mu m. Tension and [Ca2+](i) were recorded during the slow
decline of tension following stimulation in the presence of cyclopiaz
onic acid (CPA), a sarcoplasmic reticulum Ca2+-uptake pump inhibitor.
[Ca2+](i) was estimated by injecting the K+ salt form of the fluoresce
nt dye fura-2 into the fibres. Experimental temperature was 3.0 degree
s C. 2. At a SL of 2.2 mu m, where thick and thin filaments fully over
lap, the [Ca2+](i) corresponding to 50% tension generation ([Ca2+](50)
) was 1.09 +/- 0.02 mu M (mean +/- S.E.M., n = 61 contractions). At a
SL of 2.9 mu m, where overlap is approximately 50 %, the [Ca2+](50) wa
s significantly lower, 0.69 +/- 0.02 mu M (n = 22 contractions). This
is in agreement with previous results from skinned fibres. 3. The rela
tionship between tension and [Ca2+](i) was very steep, as reported pre
viously from experiments at a XL of 2.2 mu m in which the membrane per
meant acetoxymethyl ester form of fura-2 was used. The fall in tension
from 90 to 10% occurred in 0.12 +/- 0.01 pCa units (mean +/- S.E.M.,
n = 61) for a SL of 2.2 mu m and 0.17 +/- 0.01 pCa units (n = 22) for
a XL of 2.9 mu m, corresponding to Hill coefficients of 15.4 and 10.9,
respectively 4. We conclude that the increase in sensitivity of tensi
on to [Ca2+] that occurs in skinned skeletal muscle fibres upon stretc
h also occurs in intact fibres, that the steepness of the relation bet
ween tension and [Ca2+](i) in intact fibres reported previously cannot
be attributed to the use of the acetoxymethyl eater form of fura-2 to
report [Ca2+](i), and that the steepness decreases as myofilament ove
rlap decreases.