Calcium uptake by intestinal endoplasmic reticulum was determined during maturation in the rat. Calcium uptake was enhanced severalfold by the presence of ATP in suckling, weanling, and adolescent rats. Uptake values were higher during early life and decreased gradually with age. Calcium uptake represented transport into the intravesicular space of the microsomes as evident by marked decrease in the uptake of 0 degrees C compared with values at 25 degrees C and by rapid release of intravesicular calcium by the ionophore A23187. Calcium uptake was dependent on magnesium and media pH and was inhibited by vanadate. Sodium oxalate enhanced calcium uptake. Oligomycin and sodium azide did not inhibit calcium uptake by microsomes, suggesting that calcium uptake represents a property of the microsomes rather than mitochondrial uptake. Initial rate uptake was linear up to 30 s. Maximal uptake occurred at pH 7.2. Kinetic studies revealed a high-affinity, high-capacity system in microsomes from suckling rats (Vmax 2.26 +/- 0.2 nmol.mg protein-1.15 s-1 and Km 0.56 +/- 0.01 microM) compared with a low-capacity system in microsomes from adolescent rats (Vmax 0.72 +/- 0.1 nmol.mg protein-1.15 s-1 and Km 0.69 +/- 0.02 microM). These findings suggest that the endoplasmic reticulum of the enterocyte may play a major role in regulating intestinal cytosolic calcium homeostasis during early development.

The characteristics of phosphate transport across intestinal basolateral membranes of the rat were determined by using enriched preparations in which uphill Na+-dependent D-glucose transport could not be demonstrated, but ATP-dependent Ca2+ transport was present. Phosphate transport was saturable, Na+-dependent and exhibited Michaelis-Menten kinetics. Vmax. was 51.1 +/- 4.2 pmol/10 s per mg of protein and Km was 14 +/- 3.9 microM. The transport process was electroneutral. Tracer-exchange experiments and counter-transport studies confirmed the presence of a Na+-Pi carrier at the basolateral membrane. The presence of inside-positive membrane potential did not enhance phosphate uptake, indicating that the Na+ effect is secondary to the presence of the Na+-Pi carrier rather than an induction of positive membrane potential. The stoichiometry of this carrier at pH 7.4 was 2 Na+:1 phosphate, as shown by direct studies utilizing the static-head method. These studies are the first to determine the presence of a phosphate carrier at the basolateral membrane.

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This study characterizes for the first time, by use of a well-validated technique, glutamine transport across human basolateral membrane vesicles. Glutamine transport represented uptake into an osmotically active intravesicular space without significant metabolism. Glutamine uptake was temperature- and pH-dependent with maximal uptake at pH 7.5, and it was mediated by sodium-dependent and -independent processes. The initial rate of uptake was linear up to 20 s, as depicted by the formula gamma (nmol/mg protein) = 0.0009 X (s) - 0.0011 (r = 0.99). Kinetic analysis of glutamine uptake at concentrations between 0.01 and 0.3 mmol/L at 5 s under sodium and potassium gradients showed a maximal transport capacity (Vmax) of 0.39 +/- 0.04 and 0.21 +/- 0.02 nmol.mg protein-1.5 s-1 for sodium-dependent and -independent processes, respectively (p less than 0.01). Km values were 0.17 +/- 0.04 and 0.06 +/- 0.2 mmol/L, respectively (p less than 0.05). Glutamine uptake under the sodium-gradient condition was electrogenic whereas under the potassium-gradient it was electroneutral. Neutral amino acids inhibited both sodium-dependent and -independent processes. This study confirms and characterizes the presence of carrier-mediated glutamine uptake at the basolateral membranes of human enterocytes.