Diabetes in rat pregnancy alters renal calcium and magnesium reabsorption and bone formation in adult offspring.

AIMS/HYPOTHESIS: We tested the hypothesis that diabetes in pregnancy can result in the in-utero reprogramming of renal calcium and magnesium handling and of bone formation in the offspring, which persists into adulthood. METHODS: Male offspring of streptozotocin-treated diabetic rats (OD rats) and of control non-diabetic animals (OC rats) were investigated as neonates and at 8, 12 and 16 weeks of age. RESULTS: Compared with OC rats, urinary calcium and magnesium output was significantly reduced in OD rats at every age studied; Na+ and K+ outputs were unaffected. The renal expression of proteins involved in the tubular reabsorption of calcium (calcium ATPase, calbindin-D28k and epithelial calcium channel) was increased in OD animals compared with that in OC animals. Additionally, we observed that adult OD rats had lower trabecular and higher cortical femoral bone volumes, explained by deposition of bone on the endosteal surface. CONCLUSIONS/INTERPRETATION: These data show that diabetes in pregnancy has profound effects on male offspring in terms of renal tubular calcium and magnesium reabsorption and the normal pattern of bone formation. These effects persist into adulthood. Such long-lasting effects of diabetes on kidney and the skeleton were not suspected and could have important implications for the health of children born to diabetic women.

Failure of short-term gamma-linolenic acid treatment to reduce urinary calcium loss of diabetic rats.

Calcium re-absorption in the kidney is impaired in streptozotocin (STZ) diabetic rats, thereby causing hypercalciuria. Increased calcium loss starts within 1-2 days after induction of diabetes and reaches a plateau after 2 weeks. The excessive calcium excretion was previously shown to be reduced by treatment with gamma-linolenic acid (GLA) or evening primrose oil rich in GLA. However, in these studies, the animals were pre-treated for several weeks before injection of STZ. In the present study we investigated whether GLA can reduce calcium excretion when treatment starts at the same time as induction of diabetes. Rats were made diabetic with 60 mg/kg STZ and at the same time food was fortified with 0.4% GLA for the treatment group. A control group was treated with vehicle alone and given standard feed only. Urine was collected from animals in metabolism cages every 3rd day for a period of 26 days. The diabetic group increased their food and water consumption, and urine and faeces production as compared to the control group. The urinary loss of Ca, Mg, Zn, Na, K and creatinine was markedly increased in the diabetic group as compared to the control. GLA treatment, however, did not affect any of these variables. Analysis of fatty acids in kidneys of the rats showed an increased concentration of GLA in the treated group as compared to the two non-treated groups. We conclude that GLA treatment must commence before STZ injection in order to attenuate diabetes-induced hypercalciuria.

Localization of the nephron site of gentamicin-induced hypercalciuria in the rat: a micropuncture study.

In vivo renal micropuncture techniques were used to locate the nephron site of hypercalciuria induced by acute gentamicin infusion in anaesthetized Sprague Dawley rats. Three series of experiments were conducted. The effect of gentamicin on calcium reabsorption in the proximal tubule (Series I) and loop of Henle (Series II) was investigated using in vivo microperfusion whereas the effect on distal calcium handling (Series III) was studied using in vivo microinfusion. In all three experimental series, acute systemic gentamicin infusion at 0.28 mg kg(-1) min(-1) caused significant hypercalciuria within 30 min of commencing drug infusion. Gentamicin had no effect on the rates of urine flow or sodium excretion. Acute gentamicin infusion had no effect on unidirectional calcium reabsorption in the proximal tubule or loop of Henle despite a simultaneous and highly significant hypercalciuria at the whole kidney level. Net fluid reabsorption was also unaffected by the drug in these nephron segments. Acute gentamicin infusion significantly increased the urinary recovery of calcium following microinfusion into early distal tubules, whereas urinary calcium recovery was decreased after microinfusion into late distal tubules. We conclude that acute gentamicin-induced hypercalciuria is mediated by a decrease in calcium reabsorption in the early distal tubule. Thus, the acute hypercalciuric effect of gentamicin occurs at a different nephron site to the nephrotoxic effects associated with longer-term administration of the drug. It is, therefore, unlikely that gentamicin-induced hypercalciuria is involved in the pathogenesis of subsequent proximal tubular cell injury.

Altered calbindin mRNA expression and calcium regulating hormones in rat diabetic pregnancy.

Offspring of rats with diabetes mellitus are at risk of reduced calcium and bone mineral content. Altered expression of the maternal calcium binding proteins, calbindin-D(9K) and calbindin-D(28K), which are involved in renal and placental calcium transport, may underlie these problems.We have investigated the effect of diabetes on circulating concentrations of regulatory hormones with respect to calbindin-D mRNA concentrations. Three rat groups were studied; control (CP), streptozotocin-induced diabetic (DP), and insulin-treated diabetic (DPI) pregnant rats. Calbindin-D(9K) and calbindin-D(28K) mRNA abundance in placenta and maternal kidney were measured at days 7, 15, 18 and 21 of gestation, together with serum or plasma concentrations of 1,25 dihydroxyvitamin D(3) (1, 25(OH)(2)D(3)), parathyroid hormone (PTH), PTH-related protein (PTHrP), calcitonin, oestradiol and IGF-I. An increase in placental calbindin-D(9K) mRNA abundance between days 18 and 21 in CP and DPI rats was severely blunted in the DP rats. In contrast, renal calbindin-D(28K) mRNA abundance was greater at days 7, 15 and 18 in DP compared with CP rats, as was calbindin-D(9K) at day 18. Calcitonin concentrations showed no differences between the groups, and both PTH and IGF-I were reduced over the first half of gestation, unlike the calbindins. In contrast, the concentrations of PTHrP and 1,25(OH)(2)D(3) were reduced at term in the DP group compared with the other two groups. Plasma oestradiol concentrations were lower in DP than in CP rats at days 7, 15 and 18, and most striking was the absence in DP rats of the peak of oestradiol seen at day 18 in CP rats. Despite the similarity between changes in placental calbindin mRNA and 1,25(OH)(2)D(3), previous work has shown placental calbindin-D(9K) regulation to be vitamin-D-independent. These studies produce suggestive evidence, therefore, that PTHrP and oestradiol may be involved in the altered calbindin-D expression by kidney and placenta in rat diabetic pregnancy.

Renal function in chronically catheterized conscious diabetic rats using constant and servo-controlled infusion.

1. Infusion experiments were performed on chronically catheterized conscious rats to assess kidney function before and after the induction of diabetes mellitus with streptozotocin. 2. Two infusion regimens were used, a conventional constant-infusion protocol and a novel computer-driven, servo-controlled fluid replacement technique. The latter enables body fluid status to be maintained throughout a study occasion by servo-controlled replacement of spontaneous urinary fluid losses. 3. The chronically catheterized conscious rat infused using a servo-controlled system appears to be the optimum model for a study of diabetic renal function. The conscious preparation circumvents problems associated with anaesthesia and acute surgery. The servo-controlled infusion protocol maintains the altered fluid status of the diabetic condition. Both hyperfiltration and polyuria, characteristics of human diabetes often absent in anaesthetized and/or constantly infused diabetic rats, were seen in all conscious servo-controlled diabetic animals. 4. The new regimen enables a more accurate assessment of renal function in experimental diabetes than with previous protocols. It should prove useful in future studies, particularly those assessing the role of anti-diabetic drugs on the kidney.

Bone mineral density and composition in rat pregnancy: effects of streptozotocin-induced diabetes mellitus and insulin replacement.

A disturbed calcium homeostasis characterizes diabetic pregnancy. This study documents changes in bone mineral composition in diabetic pregnant rats and examines the effect of insulin replacement. Control pregnant (CP), diabetic pregnant (DP) and insulin-treated DP (DPi) rats were assessed for femoral calcium and magnesium content, bone mineral density (BMD) and the ratio of hypertrophic to maturing and proliferative cells in the femoral growth plate. DP rats showed a significantly (P < 0.01) lower body weight, femoral weight and length than CP rats. Femoral calcium and magnesium content was also significantly (P < 0.05) lower in DP rats, as was ash weight. When calcium and magnesium were normalized for ash weight no significant differences were apparent. A significantly (P < 0.05) lower total BMD at the distal femur was seen in DP rats. This comprised a significantly (P < 0.01) lower trabecular BMD with no significant change in cortical BMD. A significantly (P < 0.05) higher ratio of hypertrophic to maturing and proliferative cells of the femoral growth plate was evident in DP animals. DPi rats showed normal blood glucose concentrations and femoral growth plate histology. DPi rats also showed normal femoral weight and length but only partially restored femoral ash weight and mineral content. Insulin failed to normalize total or trabecular BMD. Diabetes mellitus clearly has a marked effect on bone growth and mineral content in pregnancy which may be relevant to overall calcium homeostasis. The lower bone growth, bone calcium content and trabecular BMD may be unfortunate consequences of the marked hypercalciuria reported elsewhere in diabetes and may serve to maintain normocalcaemia in the disease.

Effect of experimental diabetes mellitus on gentamicin-induced acute renal functional changes in the anaesthetized rat.

1. Rats with streptozotocin (STZ) diabetes are protected from gentamicin (GEN) nephrotoxicity. Because the chronic renal damage from GEN is preceded by acute renal functional changes (notably hypercalciuria), the present study aims to determine whether diabetes may also protect against the acute effects of the drug. If there is a link between the rapid physiological actions of GEN and its subsequent nephrotoxicity, the former may also be affected by the diabetic condition. 2. Standard renal clearance techniques were performed on anaesthetized rats that had been injected with STZ or vehicle 2 weeks previously. All animals were infused with 0.9% NaCl for 5 h and then either GEN (0.28 mg/kg per min) or 0.9% NaCl alone for 2 h. 3. Baseline fractional calcium excretion (FE(Ca)) of diabetic rats was three-fold that of control animals (6.6+/-0.2 vs 2.2+/-0.2%, respectively; P<0.01, MANOVA). Following GEN infusion, a comparable increase in FE(Ca) occurred in control and diabetic rats (5.3+/-0.6 vs 5.3+/-0.8%, respectively; NS). 4. Streptozotocin diabetes, therefore, does not alter the acute hypercalciuric response to GEN. This may suggest that the acute effects of GEN on renal calcium handling do not contribute to the subsequent nephrotoxicity. However, the higher baseline FE(Ca) seen in diabetic rats may afford protection against the renal injury caused by gentamicin.

Acute gentamicin-induced hypercalciuria and hypermagnesiuria in the rat: dose-response relationship and role of renal tubular injury.

1. Standard renal clearance techniques were used to assess the dose-response relationship between acute gentamicin infusion and the magnitude of hypercalciuria and hypermagnesiuria in the anaesthetized Sprague-Dawley rat. Also investigated were whether these effects occurred independently of renal tubular cell injury. 2. Acute gentamicin infusion was associated with a significant hypercalciuria and hypermagnesiuria evident within 30 min of drug infusion. The magnitude of these responses was related to the dose of drug infused (0.14-1.12 mg kg(-1) min[-1]). Increased urinary electrolyte losses resulted from a decreased tubular reabsorption of calcium and magnesium. 3. A rapid dose-related increase in urinary N-acetyl-beta-D-glucosaminidase (NAG) excretion was also observed in response to gentamicin infusion. However, there was no evidence of renal tubular cell injury and no myeloid bodies were observed within the lysosomes of the proximal tubular cells. Gentamicin may thus interfere with the mechanisms for cellular uptake and intracellular processing of NAG causing increased NAG release into the tubular lumen. 4. The absence of changes in renal cellular morphology indicates that the excessive renal losses of calcium and magnesium were an effect of gentamicin per se and not the result of underlying renal tubular injury. The renal effects described in this paper were apparent after administration of relatively low total drug doses, and with plasma concentrations calculated to be within the clinical range. These findings suggest that disturbances of plasma electrolyte homeostasis could occur in the absence of overt renal injury in patients receiving aminoglycoside antibiotics.

Dietary essential fatty acid supplementation, urinary calcium excretion and reproductive performance in the diabetic pregnant rat.

Hypercalciuria may be a contributory factor to the disturbed calcium homoeostasis seen in diabetic pregnant rats and their offspring. In diabetes, essential fatty acid metabolism is impaired. We have therefore investigated whether feeding a diet supplemented with essential fatty acids will ameliorate the hypercalciuria of diabetic pregnancy and improve reproductive performance. Female rats were fed a standard rat diet, a fat-free diet plus evening primrose oil or a fat-free diet plus sunflower oil. They were injected with streptozotocin or vehicle and mated. Urine samples were analysed for calcium before injection and during gestation. Term-pregnant diabetic rats fed evening primrose oil showed a 73% reduction in urinary calcium output compared with similar rats fed standard diet (P < 0.001). The corresponding reduction was 44% in diabetic rats fed sunflower oil (P < 0.001). A depletion of essential fatty acids in diabetes may therefore be associated with hypercalciuria; dietary supplementation, particularly with evening primrose oil, appears to correct the problem. Diabetic pregnant rats fed evening primrose oil showed a significantly greater live fetal mass (85 +/- 2 vs 33 +/- 12 g; P < 0.05) compared with similar rats fed standard diet. Such findings may imply a normalization of placental transport by essential fatty acids. Rats fed evening primrose, but not sunflower oil, also showed a reduced incidence of diabetes after streptozotocin injection compared with rats fed standard diet (63 vs 86%). Rats fed on evening primrose oil that did become diabetic were less hyperglycaemic than those on the standard diet (29 +/- 2 vs 37 +/- 2 mmol/l), suggesting that the oil may have anti-diabetic properties.

The effect of diabetes mellitus on urinary calcium excretion in pregnant rats and their offspring.

The effect of maternal diabetes mellitus on renal calcium excretion in pregnant rats and their offspring has been examined in order to ascertain the role of the kidney in the disturbed calcium homeostasis of infants born to diabetic mothers. Diabetic pregnant (DP) rats exhibited severe hypercalciuria which greatly exceeded the urinary calcium losses (UCaV) in non-diabetic pregnant (CP) or non-pregnant diabetic (D) rats. Means +/- S.E.M. for UCaV at day 21 (mmol/24 h) were: DP = 1.12 +/- 0.09 (n = 7); CP = 0.06 +/- 0.01 (n = 7); D = 0.63 +/- 0.06 (n = 7) (P < 0.001 DP vs CP and DP vs D). The profile for urinary calcium excretion in the three groups was different from that of other measured ions. The degree of natriuresis, for example, was comparable in DP and D rats at all stages studied. Although magnesium output was significantly greater in DP than D rats on days 14 and 21, this appeared to result from an additive effect of the magnesiuresis seen when pregnancy and diabetes were studied separately. The marked renal calcium wasting of diabetic pregnancy will have implications for overall calcium balance in the mother. For example, an enhanced intestinal calcium absorption was seen in DP rats in the second half of gestation. Means +/- S.E.M. for day 21 (mmol/24 h) were: DP = 3.8 +/- 0.8 (n = 7); CP = 1.4 +/- 0.3 (n = 7); D = 1.6 +/- 0.3 (n = 7) (P < 0.05 DP vs CP and DP vs D).(ABSTRACT TRUNCATED AT 250 WORDS)

Plasma sialic acid in animal models of diabetes mellitus: evidence for modulation of sialic acid concentrations by insulin deficiency.

An elevated circulating sialic acid concentration is a risk factor for cardiovascular disease. Serum sialic acid levels are elevated in NIDDM but not in uncomplicated IDDM. To study why sialic acid is increased in some types of diabetes, we assayed plasma sialic acid in various animal models of diabetes: obese (ob/ob) mice, before and after streptozotocin treatment, neonatal streptozotocin-treated (nSTZ) rats, and diabetic BB rats during and after insulin treatment. In obese mice, which exhibit moderate hyperglycemia and marked hyperinsulinemia, plasma sialic acid was decreased by 45% (fed) and 42% (fasted), compared to lean controls. Fasting reduced plasma glucose and insulin but increased sialic acid in the obese and lean mice. There was a negative correlation (r = -0.84, P < 0.001) between log plasma insulin and sialic acid in the lean and obese mice. The plasma sialic acid:globulin ratio was reduced by 35% in obese mice vs. lean controls, indicating that there may be altered sialylation of glycoproteins in obese mice. Streptozotocin treatment of obese and lean mice reduced plasma insulin but increased sialic acid. In nSTZ rats, hyperglycemia was associated with mild hypoinsulinemia, but not significantly different from control animals, and sialic acid was not altered. In diabetic BB rats, plasma glucose rose from a mean of 4.9 to 23.5 mM 48 hr after insulin withdrawal but sialic acid did not change. We conclude that an elevated plasma sialic acid level is associated with marked insulin deficiency, rather than hyperglycemia per se. The magnitude and speed of this change in sialic acid varies between species.

Effect of diabetes mellitus on maternofetal flux of calcium and magnesium and calbindin9K mRNA expression in rat placenta.

The effect of maternal diabetes mellitus on placental unidirectional maternofetal flux of calcium (Ca) and magnesium (Mg), calbindin9K mRNA expression, and net fetal Ca and Mg accretion has been investigated using control (C), untreated diabetic (D(O)) and insulin-treated diabetic (DI) rats. Unidirectional maternofetal flux of Ca in the D(O) group was 61 and 63% of the value of the C and DI groups; unidirectional maternofetal flux of magnesium in the D(O) group was 79 and 66% of the value in the C and DI groups. Fetal Ca and Mg content (mmol; mean +/- SEM) was also significantly lower in the D(O) group compared with the other two groups (0.111 +/- 0.004 versus 0.153 +/- 0.008 in C and 0.168 +/- 0.007 in DI, p < 0.01 D(O) versus C and DI for Ca; and 0.021 +/- 0.001 versus 0.027 +/- 0.001 in C and 0.031 +/- 0.001 in DI, p < 0.01 D(O) versus C and DI for Mg). However, only Ca content was significantly lower in the D(O) group when normalized to fetal ash weight. Densitometric analysis of autoradiograms after Northern hybridization with cDNA probes demonstrated that the placental calbindin9K/cyclophilin mRNA ratio was 11- to 12-fold lower in the D(O) group compared with the C and DI groups. Collectively, the data suggest that untreated maternal diabetes mellitus reduces fetal Ca and Mg accretion by an effect on the expression of placental transport components involved in the maternofetal transfer of these cations.

Effects of gentamicin, neomycin and tobramycin on renal calcium and magnesium handling in two rat strains.

1. Standard renal clearance techniques were used to compare the acute effects of gentamicin, neomycin and tobramycin on renal calcium and magnesium handling in Sprague-Dawley and Fischer 344 rats. 2. Significant hypercalciuric and hypermagnesiuric responses to all three drugs (P < 0.01) were apparent within 30 min of the onset of drug infusion. 3. The magnitude of the acute hypercalciuric and hypermagnesiuric response to the three aminoglycosides was comparable. This contrasts with their nephrotoxic action where neomycin >> gentamicin > tobramycin. The magnitude of the acute physiological responses to these drugs do not therefore reflect their nephrotoxic potential. 4. Sprague-Dawley rats were at least as responsive as Fischer rats in their acute renal responses to gentamicin. If Fischer rats are more sensitive to aminoglycoside nephrotoxicity than Sprague-Dawley rats, this is not reflected in their acute responses to gentamicin.

A paired tracer microinjection technique designed for assessment of single-nephron glucose-calcium interactions in the anesthetized rat.

The first part of this study evaluates a new paired microinjection technique for studying single-nephron permeability (in this case to calcium) following injection of 5-10 nL of a Ringer solution into a superficial proximal tubule. The mean difference in fractional 45Ca recovery from two identical microinjections into the same nephron site was 2.2 +/- 0.2% for 89 paired microinjections. Individual nephrons therefore normally show differences in calcium permeability with time. However, moment-to-moment variations in ion transport in any one nephron are in a random direction; differences cancel one another out if enough experiments are performed. The technique thus appears well suited to studies where comparisons are made between the acute nephron responses to two test solutions. It specifically overcomes problems of nephron heterogeneity seen in some other micropuncture techniques. The second part of this study uses the new technique to investigate the effects of a raised intratubular D-glucose concentration on single-nephron calcium transport. Urinary 45Ca recoveries from late proximal microinjections were significantly higher when D- (as opposed to L-) glucose was included in the injectate (6.87 +/- 0.88 vs. 5.24 +/- 0.50%; p < .02). The ability of D-glucose to depress tubular calcium reabsorption at distal nephron sites may contribute to the observed hypercalciuria following systemic D-glucose loading. It may also be relevant to the acute renal failure accompanying renal stone disease, where a relationship between hypercalciuria, urolithiasis, and the consumption of refined carbohydrates has been proposed.

Effects of D-glucose, L-glucose and D-mannitol on renal calcium handling and general renal function in the rat.

Standard renal clearance techniques were used to compare the effects of D-glucose, L-glucose and D-mannitol on renal calcium handling and general renal function in the anaesthetized rat. A significant (P < 0.01) calciuresis was seen in animals infused with D-glucose. This resulted from a decreased tubular reabsorption of calcium. A similar response was not seen with L-glucose or D-mannitol. The contrasting actions of the three sugars on tubular calcium transport may relate to their different transport characteristics, since only D-glucose is actively reabsorbed. Responses of the three sugars also differed with respect to fractional fluid and sodium reabsorption which were significantly (P < 0.001) lower in animals infused with L-glucose and D-mannitol, and plasma insulin concentrations which were significantly (P < 0.01) elevated only in the D-glucose group. An unexpected finding was a highly significant (P < 0.001) and progressive fall in glomerular filtration rate (GFR) in animals infused with L-glucose. The reasons for this are not known.

Renal micropuncture study of the effects of D-glucose tubular calcium handling in the anaesthetized rat.

Tracer microinjection studies were used to grossly locate the nephron site of altered calcium transport following D-glucose loading. Superficial proximal and distal nephrons were injected with a solution containing 45Ca and either D-glucose or L-glucose (as control). Tubule calcium transport was assessed from the recovery of radioactivity in the final urine. 45Ca recoveries from early proximal microinjections were comparable in the two groups (2.63 +/- 0.68 vs. 3.70 +/- 1.19%). However, 45Ca recoveries from late proximal microinjections were significantly higher than D-glucose was included in the injectate (7.39 +/- 1.34 vs. 2.83 +/- 0.67%, P < 0.05). 45Ca recoveries from distal microinjections were also significantly higher in the D-glucose group (87.6 +/- 2.43 vs. 71.0 +/- 2.92%, P < 0.001). These data suggest that the effects of D-glucose on renal calcium handling are mediated at the level of the distal tubule, thick ascending limb of the loop of Henle and/or pars recta. The precise mechanisms involved are not known.

Gentamicin-induced hypercalciuria in the rat: assessment of nephron site involved.

Two independent techniques were used in anesthetized rats in an attempt to locate the nephron site of the reduced tubular calcium reabsorption accompanying acute gentamicin infusion. The first technique was that of lithium clearance used to assess proximal sodium (and secondarily calcium) handling. Observations that lithium clearance was comparable in control and gentamicin-treated animals (1.83 +/- 0.39 vs. 1.46 +/- 0.14 ml.min-1 for first experimental period) suggests a lack of proximal effect of the drug. The second technique was that of tracer microinjection whereby superficial nephrons were injected with 45Ca and tubule calcium transport was assessed from the recovery of radioactivity in the final urine. 45Ca recovery values from distal microinjections were comparable in control and gentamicin-treated groups (81.1 +/- 2.0 vs. 77.7 +/- 4.6%). However, 45Ca recovery values from proximal microinjections were significantly higher in the gentamicin group (9.4 +/- 1.0 vs. 3.5 +/- 0.8%; P < .001). These data suggest that the effects of gentamicin on renal calcium handling are mediated at a nephron site proximal to the distal tubule (i.e., loop of Henle or proximal tubule itself). Closer examination of individual proximal micropuncture data may point to an effect occurring predominantly in the pars recta of the proximal tubule or loop of Henle. Taken together, the results of both parts of the present study suggest that the early physiological effects of gentamicin on the kidney occur in a different nephron segment from any subsequent nephrotoxicity.

New experimental data on the relationship between diabetes mellitus and magnesium.

Diabetes mellitus is the most frequent chronic disease associated with secondary magnesium deficit. Hypomagnesaemia is a central feature of the deficit, which is often reported in experimental and clinical forms of the disease. In diabetic rats as in man, plasma magnesium concentrations may correlate inversely with the degree of hyperglycaemia. The duration of the disease also appears to be relevant. The hypomagnesaemia of diabetes might be expected to affect intracellular concentrations of the ion. However, although some animal and clinical studies have reported subnormal magnesium concentrations in blood cells, bone, and soft tissues of diabetics, the relationship between plasma magnesium concentration and intracellular level of the ion is inconsistent. Clinical studies have speculated on a potential link between the magnesium deficit of diabetes and several diabetic complications, including cardiovascular problems and retinopathy. Recent experimental studies are largely supportive of such a link; myocardial disorders associated with magnesium deficiency have been reported in diabetic mice and rabbits. It is possible that a common mechanism involving magnesium may be responsible for some of the diverse complications of diabetes. The aetiology of hypomagnesaemia in diabetes is complex. Nevertheless, plasma magnesium concentrations are ultimately determined by four processes: intake, gastrointestinal absorption, redistribution within body pools, and urinary excretion. This review considers in turn the potential role of each of these processes in the development of diabetic hypomagnesaemia. Both experimental and clinical studies suggest that hypermagnesiuria may be the major factor involved. Recent animal studies have described a specific renal tubular magnesium defect in diabetes, which, together with the osmotic diuresis, is responsible for large magnesium losses. The precise cause of the defect is unknown, but it may relate to the prolonged hyperglycaemia, insulinopenia, disturbance of phosphate metabolism, or other hormonal changes which characterize the disease.

An investigation of the acute effect of gentamicin on the renal handling of electrolytes in the rat.

Standard renal clearance techniques were used to investigate the acute effects of gentamicin on renal electrolyte handling in anesthetized rats. Data were obtained before substantial changes in tubular integrity would be expected to occur, thus permitting a distinction between the actions of the drug per se and renal changes resulting from underlying tubular damage. Infusion of gentamicin over a 3-hr period resulted in an immediate and sustained calciuresis and magnesiuresis, with no significant change in the renal clearance of sodium or potassium. Within 60 min of the onset of drug infusion at 0.28 mg/kg/min, renal calcium clearance (ml/min) increased from 0.095 +/- 0.013 to 0.210 +/- 0.016 (P less than .001) and renal magnesium clearance (ml/min) increased from 0.538 +/- 0.059 to 0.662 +/- 0.053 (P less than .01). Because it was subsequently shown that infusion of gentamicin did not alter glomerular filtration rate, it is clear that both responses resulted from a reduced tubular reabsorption of the respective ions; they were rapidly reversible when gentamicin infusion ceased. A higher dose of gentamicin (0.56 mg/kg/min for 3 hr) also significantly decreased plasma magnesium concentrations (0.40 +/- 0.01 vs. 0.49 +/- 0.02 mmol/l, treated vs. control; P less than .01). Gentamicin-induced changes in the renal handling of calcium and magnesium, therefore, occur independently of and before the development of nephrotoxicity. They may be at least partially responsible for the alteration in electrolyte homeostasis seen in humans during aminoglycoside treatment (e.g., hypomagnesemia). It is interesting to speculate on the possibility that they may also in some way contribute to the subsequent cellular injury that is known to occur with prolonged use of gentamicin.

Calcium transport in the proximal convoluted tubule and loop of Henle of rats made diabetic with streptozotocin.

In-vivo microperfusion was used to localize the reabsorptive defect responsible for the hypercalciuria of diabetes mellitus and to investigate possible causative factors. Unidirectional proximal calcium absorption was not significantly different in rats made diabetic with streptozotocin compared with controls, providing evidence against the involvement of this nephron segment in the phenomenon. Calcium absorption by the loop of Henle, was however, significantly (P less than 0.01) lower in diabetic animals (32.1 +/- 1.2 vs 40.4 +/- 0.6 pmol/min). Based on our knowledge of calcium movements within the loop, it is likely that the reabsorptive defect residues within the thick ascending limb. The calcium lesion was found to be independent of acute changes in intraluminal glucose concentration and could not be corrected by acute insulin treatment. The study also provides new information on the relationship between intratubular glucose and fluid movements in the rat nephron. In diabetic rats a proximal perfusate containing 30 mmol glucose/l resulted in fluid absorption comparable with that seen in control rats perfused with 5 mmol glucose/l. However, intraluminal glucose had a stimulatory effect on fluid absorption in the loop of Henle of diabetic rats (10.7 +/- 0.5 vs 7.9 +/- 0.4 nl/min; P less than 0.01).