[Influences of magnesium sulfate on maternal calcium metabolism in preterm labor].

The effects of intravenous magnesium sulfate tocolysis on calcium metabolism were studied in 10 patients with preterm labor. A loading dose of magnesium sulfate (4g) was administered intravenously maintenance intravenous infusion of magnesium sulfate (1g per hour). All patients simultaneously received 50 micrograms ritodrin per minutes by intravenous infusion. Serum magnesium increased from 1.91 +/- 0.06mg/dl to 4.6 +/- 0.71mg/dl at 30 minutes (p < 0.01) and it remained relatively high. The fall in serum calcium corrected by serum total protein was most rapid during the first 30 minutes, from 9.04 +/- 0.47mg/dl to 8.3 +/- 0.27mg/dl (p < 0.01). Urinary excretion of magnesium, represented as the calcium/creatinine ratio, rose markedly from 0.05 +/- 0.01 to 3.18 +/- 0.8 at an hour (p < 0.01) and thereafter remained higher than the baseline level. Changes in urinary excretion of calcium paralleled those of urinary evcretion of magnesium. Serum parathyroid hormone rose from 118 +/- 42.2pg/ml to 294 +/- 121pg/ml at 6 hours (p < 0.05). Serum 1 alpha,25-(OH)2D3-rose from 89.3 +/- 44.2pg/ml to 126 +/- 38.7pg/ml (p < 0.05). Serum calcitonin showed no significant change. These findings indicate that correction of hypocalcemia mainly depends on secretion of parathyroid hormone in the early stage, and thereafter depends on the cooperative action of parathyroid hormone and 1 alpha,25-(OH)2D3.

Identification and purification of a transverse tubule coupling protein which binds to the ryanodine receptor of terminal cisternae at the triad junction in skeletal muscle.

In fast twitch skeletal muscle, the signal for excitation-contraction coupling is transferred from transverse tubule across the triad junction; calcium is thereby released from the terminal cisternae of sarcoplasmic reticulum triggering muscle contraction. Recently, the feet structures of terminal cisternae, which bridge the gap at the triad junction, have been identified as the ryanodine receptor and in turn with the calcium release channels of sarcoplasmic reticulum. The latter consists of an oligomer of a single high molecular weight polypeptide (Mr 360,000). This study attempts to identify the component in the transverse tubule which ligands with the foot structure to form the triad junction. The purified ryanodine receptor, derivatized with sulfosuccinimidyl-2-(p-azidosalicylimido)-1,3'-dithiopropionate (SASD), a thiol-cleavable, 125I-iodinatable, and photoactive probe, was shown to selectively cross-link to a protein with Mr of 71,000 in isolated transverse tubules. This coupling protein was purified from transverse tubule by solubilization with the detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS) and then purified by sequential column chromatography. In the absence of sulfhydryl agents, the purified polypeptide has an Mr of 61,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A complementary approach using SASD was employed to confirm association of the coupling protein with the ryanodine receptor of terminal cisternae. We conclude that the transverse tubule coupling protein together with the ryanodine receptor (foot structure) is involved in the liganding between transverse tubule and terminal cisternae of sacroplasmic reticulum.

Ultrastructure of the calcium release channel of sarcoplasmic reticulum.

This study is concerned with the characterization of the morphology of the calcium release channel of sarcoplasmic reticulum (SR) from fast-twitch skeletal muscle, which is involved in excitation-contraction coupling. We have previously purified the ryanodine receptor and found it to be equivalent to the feet structures, which are involved, in situ, in the junctional association of transverse tubules with terminal cisternae of SR. The receptor is an oligomer of a single high molecular weight polypeptide and when incorporated into phospholipid bilayers, has channel conductance which is characteristic of calcium release in terminal cisternae of SR. The purified channel can be observed by electron microscopy using different methods of sample preparation, with complementary views being observed by negative staining, double staining, thin section and rotary shadowing electron microscopy. Three views can be observed and interpreted: (a) a square face which, in situ, is junctionally associated with the transverse tubule or junctional face membrane; (b) a rectangle equivalent to the side view; and (c) a diamond shape equivalent to the side view, of which the base portion appears to be equivalent to the transmembrane segment. Negative staining reveals detailed substructure of the channel. A computer averaged view of the receptor displays fourfold symmetry and ultrastructural detail. The dense central mass is divided into four domains with a 2-nm hole in the center, and is enclosed within an outer frame which has a pinwheel appearance. Double staining shows substructure of the square face in the form of parallel linear arrays (six/face). The features of the isolated receptor can be correlated with the structure observed in terminal cisternae vesicles. Sections tangential to the junctional face membrane reveal that the feet structures (23-nm squares) overlap so as to enclose smaller square spaces of approximately 14 nm/side. We suggest that this is equivalent to the transverse tubule face and that the terminal cisternae face is smaller (approximately 17 nm/face) and has larger alternating spaces as a consequence of the tapered sides of the foot structures. Image reconstruction analysis appears to be feasible and should provide the three-dimensional structure of the channel.