Calcium fluxes in mouse mammary tissue in vitro: intracellular and extracellular calcium pools

Abstract

1. The total Ca content of the mammary gland increased from about 2 to 12 mumole/g tissue during the transition from pregnancy to lactation in the mouse. In tissue from lactating mice at least two thirds of the total Ca exchanged with external Ca in 6 hr. There was little non-exchangeable Ca in tissues from pregnant mice.2. At 37 degrees C the time courses of influx and efflux of (45)Ca in lactating tissues could be analysed by assuming three exponential components with rate constants of about 0.3, 0.06 and 0.005 min(-1) and containing, respectively, 1.7, 1.5 and 4.7 mumole (45)Ca/g tissue at the steady state.3. The rapidly effluxing component showed the time- and temperature-dependence characteristic of bulk-phase-limited diffusion through the extracellular space. The diffusion coefficient was about one quarter of the self-diffusion coefficient of Ca in aqueous solution, consistent with a tortuosity factor of about 2. A portion of the Ca in this component was displaced by La(3+). The amount remaining in the presence of 3 mm-La(3+) was close to that expected for free extracellular Ca. The rapid component was therefore interpreted as originating from an extracellular compartment containing both free and bound Ca.4. The rate of efflux of the intermediate component was slowed by a factor of ten when the temperature was decreased from 37 to 0 degrees C giving a Q(10) of 2.7, expected for membrane transport. The slow component present at 37 degrees C was not displaced by EGTA or La(3+), suggesting that it is not localized extracellularly. It was not apparent in the 0 degrees C efflux curves.5. The biphasic time course of uptake of ionophore (A23187)-releasable (45)Ca in particulate fractions obtained by homogenization and centrifugation of tissues which had been incubated with the isotope was consistent with the hypothesis that the two slower components of (45)Ca flux originate from intracellular compartments. Mitochondrial uptake probably did not contribute significantly to Ca exchange in these tissues.6. (45)Calcium fluxes in mammary tissues from pregnant mice also showed three components with rate constants similar to those found in tissues from lactating mice. The amount of Ca in each component was much smaller than in lactating tissue when compared on the basis of tissue weight.7. We conclude from these studies that: (i) intra- and extracellular Ca pools in mammary tissue can be distinguished on the basis of the temperature dependence of their fluxes and (ii) the transition from pregnancy to lactation is accompanied by large increases in both intra- and extracellular Ca pools in mammary alveolar cells.