Enhancement of rat intestinal calcium absorption by vanadate.

Vanadate alters intestinal transport and may have a role in regulating cell function. To determine whether it influences calcium absorption, we tested the effects of acute and chronic vanadate administration on calcium absorption using single-pass perfusion of jejunal and ileal segments of the in vivo rat intestine. Acute vanadate administration increased the lumen-to-mucosa and net fluxes of calcium in both the jejunum and ileum. The increase was largely due to an enhancement of the saturable fluxes of calcium and was observed at 10(-4) M concentration of vanadate, but not at higher or lower concentrations of the oxyanion, except at the highest concentration used, 10(-2) M, where calcium absorption was inhibited. Chronic vanadate administration caused, on the other hand, no changes in calcium absorption. We have demonstrated previously that rat intestinal (Na+ + K+)-ATPase is inhibited by vanadate, an effect that could raise cell sodium and increase the efflux of sodium across the brush border membrane. The results suggest that the vanadate enhancement of calcium absorption may be related to an increased entry of calcium into the mucosa, possibly as a result of an augmented exchange through the Na+/Ca+ antiport system. Alternatively, vanadate may influence access to a calcium channel in the mucosal membrane of the intestinal epithelium, leading to the observed increase in absorption.     

Mechanisms of intestinal calcium absorption

Calcium is absorbed in the mammalian small intestine by two general mechanisms: a transcellular active transport process, located largely in the duodenum and upper jejunum; and a paracellular, passive process that functions throughout the length of the intestine. The transcellular process involves three major steps: entry across the brush border, mediated by a molecular structure termed CaT1, intracellular diffusion, mediated largely by the cytosolic calcium-binding protein (calbindinD(9k) or CaBP); and extrusion, mediated largely by the CaATPase. Chyme travels down the intestinal lumen in approximately 3 h, spending only minutes in the duodenum, but over 2 h in the distal half of the small intestine. When calcium intake is low, transcellular calcium transport accounts for a substantial fraction of the absorbed calcium. When calcium intake is high, transcellular transport accounts for only a minor portion of the absorbed calcium, because of the short sojourn time and because CaT1 and CaBP, both rate-limiting, are downregulated when calcium intake is high. Biosynthesis of CaBP is fully and CaT1 function is approximately 90% vitamin D-dependent. At high calcium intakes CaT1 and CaBP are downregulated because 1,25(OH)(2)D(3), the active vitamin D metabolite, is downregulated.     

Effects of experimental diabetes on intestinal strontium absorption in the rat.

Control and streptozotocin diabetic rats were studied at 5 and 12 days after induction of diabetes. Strontium absorption was measured by in situ perfusion of duodenum and ileum. Duodenal absorptive capacity (absorption per unit length) and absorptive specific activity (absorption per gram of dry weight mucosa) were depressed. Depression was present both at 5 days, when mucosal growth is similar in controls and diabetics, and at 12 days, when mucosal growth is 50% greater in diabetics. Effects of diabetes on ileal absorption were minimal in comparison with effects on duodenum. This depression of duodenal strontium absorption in the diabetic rat is analogous to effects of diabetes on calcium absorption and may be mediated by abnormal vitamin D metabolism.     

Modulation of intestinal absorption of calcium]

1. Absorption of ingested calcium (2 ml of a 10mM CaCl2 solution + 45Ca) by the adult rat was shown to be facilitated by the simultaneous ingestion of an active carbohydrate, L-arabinose. As the carbohydrate concentration is increased from 10 to 200 mM, the adsorption of calcium is maximized at a level corresponding to about twice the control adsorption level. 2. A similar doubling of calcium adsorption is obtained when a 100 mM concentration of any one of a number of other carbohydrates (gluconic acid, mannose, glucosamine, sorbitol, lactose, raffinose, stachyose) is ingested simultaneously with a 10 mM CaCl2 solution. 3. Conversely, the simultaneous ingestion of increasing doses (10 to 100 mM) of phosphate (NaH2PO4) with a 10 mM CaCl2 solution results in decreased 45Ca absorption and retention by the adult rat. 4. The maximum inhibition of calcium adsorption by phosphate is independent of the concentration of the ingested calcium solution (from 5 to 50 mM CaCl2). 5. The simultaneous ingestion of CaCl2 (10 mM) with lactose and sodium phosphate (50 and 10 mM, respectively) shows that the activating effect of lactose upon 45Ca adsorption may be partly dissimulated by the presence of phosphate. 6. These various observations indicate that, within a large concentration range (2 to 50 mM CaCl2), calcium adsorption appears to be a precisely modulated diffusion process. Calcium absorption varies (between minimum and maximum levels) as a function of the state of saturation by the activators (carbohydrates) and inhibitors (phosphate) of the calcium transport system.     

Effects of different metabolites of vitamin D3 and of calcium concentration on the intestinal absorption of strontium]

The effect of calcium concentration and vitamin B3 25 OHD3 and 1.25 diOHD3 upon intestinal strontium transport was studied in vitamin D deficient rats with duodenal perfusion in situ. When the calcium concentration was increased, the strontium passive absorption was decreased. The vitamin D3 and its derivatives increased intestinal strontium transport significantly and this increment was not modified by calcium. In our experiment, calcium is competitive with strontium only in passive absorption.     

Regulation of goby intestinal ion absorption by the calcium messenger system

The role of the calcium messenger system in the regulation of ion absorption across the teleost intestine was studied using pharmacological intervention. Radiochloride transport was independent of external Ca2+ over the range 10 microM to 2.5 mM. Treatment with the Ca2+ ionophore A23187 (to hyperpolarization of the apical membrane potential of intestinal epithelial cells. The Ca2+-calmodulin antagonists trifluoperazine (TFP) and calmidazolium (R24571) produced opposite effects, i.e., stimulation of Cl- absorption and cellular depolarization. Treatment with TFP or R24571 will block or override the inhibitory action of A23187. These data suggest a regulatory role for Ca2+ in the control of intestinal NaCl absorption and mediation via calmodulin.     

Removal of radioactive strontium from the rat by feeding stable strontium

The effect of administering the stable isotope of strontium (as phos-phate) at different dietary levels to adult rats (fed on a cereal and pulse-based diet containing 0.4% Ca) on the retention of radiostrontium (⁸⁹Sr) and radiocalcium (⁴⁵Ca) in the femur and the whole skeleton was studied for a period up to 6 weeks after an intraperitoneal injection of the two radioisotopes. The ability of strontium to remove⁸⁹Sr under the above dietary conditions was examined. Feeding Sr at 0.5% or 1% levels for 6 weeks had no effect on the skeletal content of⁸⁹Sr or⁴⁵Ca while a dietary regimen of 2% Sr (2000 times the normal content), significantly lowered the⁸⁹Sr and⁴⁵Ca content by about 30% in the femur but not in the whole skeleton. At this Sr level, the urinary excretion of the isotopes increased with a concomitant decrease in their excretion in the faeces. This study underscores the limitations of dietary Sr to mobilise⁸⁹Sr from the bones after it is incorporated in the bone mineral.     

Mechanisms and functional aspects of intestinal calcium absorption

Calcium absorption, in terms of mechanisms and function, is well adapted to meet the calcium needs of mammals. When calcium levels in the food are low, the active, mediated transcellular calcium transport assumes primary importance. This process is vitamin D-dependent, largely localized in the duodenum, and involves three steps: entry across the brush border, mediated by a molecular structure, CaT1, with two components; a facilitated transport that saturates at low luminal calcium concentration; and a channel component through which most calcium enters the cell at the higher luminal concentrations. Intracellular diffusion is assured by a small, cytosolic calcium binding molecule, calbindinD(9k), which carries more than 90% of the calcium that traverses the duodenal cell, thus also serving as a buffer. Extrusion is by the CaATPase and is not a limiting step. Calcium entry is reduced by more than 90% in the absence of vitamin D, with biosynthesis of calbindinD(9k) totally vitamin D-dependent. Active transport is upregulated on low calcium intake and downregulated at high calcium intake, when paracellular calcium transport through the tight junctions of the intestine becomes the dominant process. The amount of calcium absorbed paracellularly is a function of the calcium gradient between lumen and plasma and of the time the chyme spends at a given intestinal site. The coexistence of mediated and nonmediated transport processes assures the organism of an adequate calcium supply, yet prevents excessive calcium absorption.     

Determining the sources of calcium for migratory songbirds using stable strontium isotopes

We investigated natural variations in the stable isotopic composition of strontium (a surrogate for calcium) in the bones of a single species of breeding migratory songbird, as well as in their eggshells, egg contents, and food sources. We use this information to determine the sources of calcium to these migratory songbirds and their offspring. Samples were collected from two locations in the northeastern USA (Hubbard Brook, NH, and Downer Forest, VT.) that differed in soil geochemistry. The mean ⁸⁷Sr/⁸⁶Sr ratios of food items (caterpillars and snails), eggshells, and egg contents were indistinguishable within each site, but significantly different between the two sites. Mean ⁸⁷Sr/⁸⁶Sr ratios for the bones of adult females were significantly different between the two sites, but values were significantly lower than those of food items and eggshells at each site. Two of four adult individuals studied at each site had ⁸⁷Sr/⁸⁶Sr ratios lower than the entire range of values for local food sources. Mixing calculations indicate that up to 60% of skeletal strontium and calcium was derived from foods consumed in the winter grounds where lower ⁸⁷Sr/⁸⁶Sr ratios predominate. At each study site, the ⁸⁷Sr/⁸⁶Sr ratio of eggshells differed significantly between clutches, but the mean clutch ⁸⁷Sr/⁸⁶Sr ratios were unrelated to the skeletal ⁸⁷Sr/⁸⁶Sr ratio of the laying adult. These findings suggest that strontium (and hence calcium) for eggshell production in this species is derived predominantly from local food sources in breeding areas. Thus, reductions in available calcium in northern temperate ecosystems due to the influences of acid deposition could be potentially harmful to this and other species of migratory bird.     

Effect of dietary calcium and phosphorus on intestinal calcium absorption and vitamin D metabolism.

To understand better dietary regulation of intestinal calcium absorption, a quantitative assessment of the metabolites in plasma and duodenum of rats given daily doses of radioactive vitamin D₃ and diets differing in calcium and phosphorus content was made. All known vitamin D metabolites were ultimately identified by high-pressure liquid chromatography. In addition to the known metabolites (25-hydroxyvitamin D₃, 24,25-dihydroxyvitamin D₃, 1,25-dihydroxyvitamin D₃, 25,26-dihydroxyvitamin D₃, and 1,24,25-trihydroxyvitamin D₃), several new and unidentified metabolites were found. In addition to 1,25-dihydroxyvitamin D₃ and 1,24,25-trihydroxyvitamin D₃, the levels of some of the unknown metabolites could be correlated with intestinal calcium transport. However, whether or not any of these metabolites plays a role in the stimulation of intestinal calcium absorption by low dietary calcium or low dietary phosphorus remains unknown.     

Chromaffin cell calcium channel kinetics measured isotopically through fast calcium, strontium, and barium fluxes

Fast Ca2+ uptake into K+-depolarized cultured bovine adrenal chromaffin cells has been isotopically measured in a time scale of 1-10 s. Depolarized cells retained as much as 80-fold 45Ca2+ taken up by resting cells; Ca2+ was not taken up by fibroblasts or endothelial-like cells. Because Ca2+ entry was inhibited by inorganic (La3+, Co2+, Mg2+) and organic (nifedipine) Ca2+ channel antagonists and enhanced by the Ca2+ channel activator Bay-K-8644, it seems clear that Ca2+ gains access to the chromaffin cell cytosol mainly through specific voltage-dependent Ca2+ channels. Ca2+ uptake evoked by 59 mM K+ was linear during the first 5 s of stimulation and continued to rise at a much slower rate up to 60 s. The rate of Ca2+ entry became steeper as the external [Ca2+] increased; initial rates of Ca2+ uptake varied from 0.06 fmol/cells . s at 0.125 mM Ca2+ to 2.85 fmol/cell . s at 7.5 mM Ca2+. The early 90Sr2+ uptake was linear but faster than Ca2+ uptake and later on was also saturated; 133Ba2+ was taken up still at a much faster rate and was linear for the entire depolarization period (2-60 s). Increased [K+] gradually depolarized chromaffin cells; Ca2+ and Sr2+ uptakes were not apparent below 30 mM K+ but were linear for 30 to 60 mM K+. In contrast, substantial Ba2+ uptake was seen even in K+-free solutions; and in 5.9 mM K+, Ba2+ uptake was as high as Ca2+ uptake obtained in 60 mM K+. Five to ten-second pulses of 45Ca2+, 90Sr2+, or 133Ba2+ given at different times after pre-depolarization of chromaffin cells served to analyze the kinetics of inactivation of the rates of entry of each divalent cation. Inactivation of Ca2+ uptake was faster than Sr2+, and Ba2+ uptake inactivated very little. Neither voltage changes nor Ca2+ ions passing through the channels seems to cause their inactivation; however, experiments aimed to manipulate the levels of internal Ca2+ using the cell-permeable chelator Quin-2 or the ionophore A23187 strongly suggest that intracellular Ca2+ levels determine the rates of inactivation of these channels.     

Effects of calcium and sugars on intestinal manganese absorption

An in vivo luminal perfusion technique was used to investigate the influence of Ca, Mg, lactose, and glucose on Mn absorption in different segments of the rat intestine. Mn absorption was determined by measuring disappearance of⁵⁴Mn activity from the perfusion solution containing 0.1 or 0.01 mmol/L Mn. Na and water absorption were also determined. Mn absorption decreased during the first 30 min of perfusion to reach a steady state thereafter. Ca (1 mmol/L) inhibited Mn absorption in the proximal jejunum and in the colon, whereas Mn absorption was increased by Ca in the distal jejunum. Mg (1 mmol/L), lactose, and glucose (25 mmol/L each) had no effect on Mn absorption in the jejunum. These results can be explained by a direct interaction of Mn and Ca during transcellular Ca transport in the proximal jejunum and colon. The reason for the stimulatory effect of Ca in the distal jejunum is unknown.     

Control of calcium absorption and intestinal calcium-binding protein synthesis

A current hypothesis suggests that the degree of Ca absorption is hormonally controlled via the feed-back regulation of 1,25-dihydroxycholecalciferol (1,25-(OH)₂D₃) production from 25-hydroxycholecalciferol (25-OHD₃) by kidney 1-hydroxylase. To test this hypothesis, dihydrotachysterol₃ (DHT₃), a steroid not requiring 1-hydroxylation for biological activity, was given to chicks as the only source of vitamin D-activity. As expected, DHT₃-treated chicks did not adapt to a calcium-deficient diet. However, both the efficiency of Ca absorption and net synthesis of CaBP were stimulated in DHT₃-treated chicks by a low phosphorus intake, providing evidence for an alternate pathway of control.     

Antioxidant and antiapoptotic properties of melatonin restore intestinal calcium absorption altered by menadione

The intestinal Ca²⁺ absorption is inhibited by menadione (MEN) through oxidative stress and apoptosis. The aim of this study was to elucidate whether the antioxidant and antiapoptotic properties of melatonin (MEL) could protect the gut against the oxidant MEN. For this purpose, 4-week-old chicks were divided into four groups: (1) controls, (2) treated i.p. with MEN (2.5 μmol/kg of b.w.), (3) treated i.p. with MEL (10 mg/kg of b.w.), and (4) treated with 10 mg MEL/kg of b.w after 2.5 μmol MEN/kg of b.w. Oxidative stress was assessed by determination of glutathione (GSH) and protein carbonyl contents as well as antioxidant enzyme activities. Apoptosis was assayed by the TUNEL technique, protein expression, and activity of caspase 3. The data show that MEL restores the intestinal Ca²⁺ absorption altered by MEN. In addition, MEL reversed the effects caused by MEN such as decrease in GSH levels, increase in the carbonyl content, alteration in mitochondrial membrane permeability, and enhancement of superoxide dismutase and catalase activities. Apoptosis triggered by MEN in the intestinal cells was arrested by MEL, as indicated by normalization of the mitochondrial membrane permeability, caspase 3 activity, and DNA fragmentation. In conclusion, MEL reverses the inhibition of intestinal Ca ²⁺ absorption produced by MEN counteracting oxidative stress and apoptosis. These findings suggest that MEL could be a potential drug of choice for the reversal of impaired intestinal Ca ²⁺ absorption in certain gut disorders that occur with oxidative stress and apoptosis.