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  • Title: Effects of internal sodium and hydrogen ions and of external calcium ions and membrane potential on calcium entry in squid axons.
    Author: Mullins LJ, Tiffert T, Vassort G, Whittembury J.
    Journal: J Physiol; 1983 May; 338():295-319. PubMed ID: 6875960.
    Abstract:
    Squid giant axons were impaled with electrodes to measure pNai, pHi, Em, and were injected with either aequorin or arsenazo III to measure [Ca]i or with phenol red to measure [H]i. Depolarization of such axons with elevated [K] in sea water leads to a Ca entry that is a function of [Ca]o, [Na]i, and [H]i. With saturating [Na]i half-maximal Ca entry is produced by a [Ca]o of 0.58 mM. With saturating [Ca]o, depolarization produced by 450 mM-K+ leads to half-maximal Ca entry when [Na]i is 25 mM; entry is virtually undetectable if [Na]i is 18 mM. If [Ca]o is 50 mM, Ca entry upon depolarization as measured with aequorin is phasic with a rapid phase of light emission and a plateau; Ca entry as measured with arsenazo III shows no such phasic behaviour, absorbance vs. time is a square wave that closely follows the depolarization vs. time trace. Both detectors of [Ca]i show a square-wave response if [Ca]o is 3 mM. The introduction of 2 mM-CN into the sea water bathing the axon does not affect the response to depolarization nor does the destruction of most of the ATP in the axon following the injection of apyrase. If axons are microinjected with phenol red rather than arsenazo, the entry of Ca produces an acidification in the peripheral parts of the axoplasm. Other experiments measuring [Ca]i show that Ca entry is strongly inhibited by a decrease in pHi. Making sea water alkaline with pH buffers scarcely affects the Ca entry induced by depolarization; making axoplasm alkaline by adding NH4+ to sea water greatly enhances Ca entry by Na/Ca exchange and also enhances the ability of axoplasmic buffers to absorb Ca.
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