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[ATP-dependent Ca2+-uptake by the plasma membrane fraction of the myometrium]
Abstract:
The specific activities of Mg2+, Ca2+-ATPase in the plasma membrane fraction of rabbit and cattle myometrium are 8.30 +/- 0.80 and 2.36 +/- 0.48 mkmoles of Pi per mg of protein, respectively. This fraction possesses a higher (in comparison with other subcellular fractions) capacity for ATP-dependent uptake of 45Ca2+ (9.37 +/- 1.66 and 6.86 +/- 0.96 nmoles of 45Ca2+ per mg of protein in 15 min for rabbit and cattle myometrium, respectively); the ratio of ATP-dependent uptake of Ca2+ to adsorbed Ca2+ is also high. Phosphate increases Ca2+ uptake in the presence of ATP and Mg2+. The ionophore A-23187 added to the incubation mixture without ATP and Mg2+ sharply increases Ca2+ binding. An addition of the ionophore at the 15th min of the ATP-dependent Ca2+ uptake causes a complete and rapid release of the accumulated Ca2+. The release of Ca2+ can be also caused by an addition of Na-DS or EGTA to the incubation mixture. This suggests that Ca2+ is accumulated through the plasma membrane inside the closed structures. It was assumed that myometrial sarcolemma plays an essential role in regulation of intracellular Ca2+ concentration in the uterus at rest and that the active Ca2+ efflux from the cells is controlled by the Mg2+, Ca2+-ATPase system.
Insights
The myometrial plasma membrane
Area of Science:
- Biochemistry
- Cell Physiology
Background:
- The myometrium's role in uterine function is critical.
- Intracellular calcium (Ca2+) concentration is a key regulator of myometrial contractility.
Purpose of the Study:
- To investigate the role of Mg2+, Ca2+-ATPase in the myometrial plasma membrane.
- To understand the mechanisms of Ca2+ uptake and release in myometrial cells.
Main Methods:
- Isolation of plasma membrane fractions from rabbit and cattle myometrium.
- Assay of Mg2+, Ca2+-ATPase activity.
- Measurement of ATP-dependent 45Ca2+ uptake and release.
- Use of ionophore A-23187, Na-DS, and EGTA.
Main Results:
- Plasma membrane fractions exhibited significant Mg2+, Ca2+-ATPase activity.
- High capacity for ATP-dependent Ca2+ uptake was observed in the plasma membrane.
- Phosphate enhanced Ca2+ uptake, while ionophore A-23187 facilitated Ca2+ release.
- Ca2+ accumulation occurred within closed structures, suggesting sarcolemmal involvement.
Conclusions:
- The myometrial sarcolemma plays a vital role in regulating intracellular Ca2+ concentration.
- The Mg2+, Ca2+-ATPase system is crucial for controlling active Ca2+ efflux from myometrial cells.