Brinley F J, Spangler S G, Mullins L J
J Gen Physiol. 1975 Aug;66(2):223-50. doi: 10.1085/jgp.66.2.223.
Ca efflux in dialyzed squid axons was measured with 45Ca as a function of internal ionized Ca in the range 0.005-10 muM. Internal Ca stores were depleted by treatment with CN and dialysis with media free of high energy compounds. The [Ca]iota was stabilized with millimolar concentrations of EDTA, EGTA, or DTPA. Nonspecific leak of chelated Ca was measured with [14C]-EDTA and found to be 0.02 pmol/cm2s/mM EDTA. Correction of the measured Ca efflux for this leak of chelated calcium was made when appropriate. Ca efflux was roughly linear with internal free Ca in the range 0.005-0.1 muM. Above 0.1 muM, efflux was less than proportional to concentration but did not saturate at the highest concentration studied. Ca efflux was reduced about 50% by replacement of external Na with Li at Caiota approximately 1 muM, but was insensitive to such replacement for Ca less than 0.1 muM. Ca efflux was insensitive to internal Mg in the range 0-4 mM, indicating that the Ca pump favors Ca over Mg by a factor of about 10(6). Ca efflux was reduced about 60% by increasing internal Na from 1 to 80 mM. This effect could represent weak interference of a Ca carrier by Na or a loss of driving force because of a reduction in ENa - Em occasioned by an increase in Naiota. A few measurements were made of Ca influx in intact and in dialyzed fibers. In both cases, Ca influx increased when external Na was replaced by Li.
在透析的枪乌贼轴突中,以45Ca测量Ca外流,作为0.005 - 10 μM范围内内部游离Ca的函数。通过用CN处理和用不含高能化合物的介质透析来耗尽内部Ca储备。用毫摩尔浓度的EDTA、EGTA或DTPA使[Ca]i稳定。用[14C]-EDTA测量螯合Ca的非特异性泄漏,发现为0.02 pmol/cm2s/mM EDTA。在适当的时候对测量的Ca外流进行螯合钙泄漏的校正。在0.005 - 0.1 μM范围内,Ca外流与内部游离Ca大致呈线性关系。高于0.1 μM时,外流与浓度的比例减小,但在研究的最高浓度下未饱和。当Cai约为1 μM时,用Li替代外部Na可使Ca外流减少约50%,但对于小于0.1 μM的Ca,这种替代不敏感。在0 - 4 mM范围内,Ca外流对内部Mg不敏感,表明Ca泵对Ca的偏好超过Mg约10(6)倍。通过将内部Na从1 mM增加到80 mM,Ca外流减少约60%。这种效应可能代表Na对Ca载体的弱干扰或由于Nai增加导致ENa - Em降低而失去驱动力。对完整和透析纤维中的Ca内流进行了一些测量。在这两种情况下,当外部Na被Li替代时,Ca内流均增加。