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. 1979 Jan;25(1):45–62. doi: 10.1016/S0006-3495(79)85277-7

Reference Phase Analysis of Free and Bound Intracellular Solutes

II. Isothermal and Isotopic Studies of Cytoplasmic Sodium, Potassium, and Water

Samuel B Horowitz, Philip L Paine
PMCID: PMC1328447  PMID: 262386

Abstract

The intracellular reference phase (RP) method and ultra-low temperature micro-dissection were used for isothermal and isotopic phase distribution studies of Na+, K+, and water in amphibian oocyte cytoplasm. One-third of the cytoplasmic water is available as solvent for [3H]sucrose. This fraction, designated c1, quantitatively coincides with the water volume in which Na+ and K+ are freely diffusible. Two-thirds of the cytoplasmic water is inaccessible to sucrose and is designated c2. The Na+ and K+ associated with c2 are extremely slowly exchanging (bound) and at different concentrations than in c1. The cations in c1 are in mass-action equilibria with those in c2, each described by an equation of the form

Cci = Cc1i + Cc2i = qi·CRPi + maxCc2i·f(CRPi

in which Cci is the cytoplasmic Na+ or K+ concentration, Cc1i is the free, and Cc2i the bound cation concentration averaged over the cytoplasmic water. qi is the fractional free solute space, CRPi the RP concentration, maxCc2i the concentration of binding sites, and the function f is satisfied by the Langmuir isotherm. Numerical values for the variables of the isotherm are determined. Activity coefficients are calculated from RP data and provide a basis for generalizing the oocyte results to other cells. The conclusion is drawn that both c1 and c2 are widely distributed in cells, and that cellular ionic activities involve two distinct systems: the cell-membrane system and an adsorbed water ion-exchange-like buffering system. Alternative explanations for the two-component cytoplasm are considered. A model is proposed in which c1 is a normal intracellular aqueous phase controlled by the plasma membrane, whereas c2 consists of water and ions adsorbed in hydrate crystalline structures. In oocytes these structures are identified with yolk platelets.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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