Pinealocyte subsurface cisterns. III: Storage of calcium ions and their probable role in cell stimulation

Different techniques for the ultrastructural demonstration of calcium have been applied to the pineal gland of Meriones unguiculatus, attention being focussed on the endoplasmic reticulum (ER) and its subsurface cisterns (ssc). By means of a "loading" method [Walz, 1982; Wakasugi et al., 1982] it is shown that the pinealocyte ER-ssc system sequesters calcium with dependency on ATP. Furthermore, a modification of the method of Duce and Keen [1978] is presented which turned out a) to be sensitive enough to demonstrate the cell's own low amounts of calcium as fine granular precipitates, and b) to preserve ultrastructure sufficiently. This method rendered possible comparison of the calcium distribution inside pinealocytes of the following groups: animals fixed during daytime, animals fixed at night, animals fixed at night with prior exposure to bright white light, animals fixed at night but injected at the end of the preceding light period with a pharmacon known to prevent the release of calcium from the ER of muscle fibers (Dantrolen). In contrast to the daytime findings, the pinealocyte ER-ssc system at night is free of precipitable calcium; nocturnal illumination induces reacquisition, Dantrolen hinders nocturnal depletion. From the nocturnal coincidence of pinealocyte activity and calcium release from ssc, and from other cytological and experimental data, it is concluded that the functional significance of ssc refers to the regulation of pinealocyte sensitivity. Vice versa, pinealocyte activity may influence ER expansion and ssc size via the calcium-dependent stability of microtubules.