Maternal zinc restriction affects postnatal growth and glucose homeostasis in rat offspring differently depending upon adequacy of their nutrient intake.

INTRODUCTION: We have previously investigated effects of moderate maternal zinc (Zn) restriction on growth and glucose homeostasis in offspring, but interaction between maternal Zn restriction and postnatal nutrition have not been studied. RESULTS: Weight and serum Zn were lower in ZnD-IN than in ZnC-IN rats at wk 3, but ZnD-AN and ZnD-EN rats had greater weights than respective controls and higher insulin-like growth factor-1 (ZnD-AN) and leptin levels (ZnD-EN). Subsequently, both ZnD-AN and ZnD-EN pups were insulin resistant, and had evidence of elevated serum leptin and depressed insulin receptor phosphorylation with gender-specific differences up to 15 weeks. DISCUSSION: Maternal Zn restriction interacted with postnatal nutritional status, resulting in divergent effects on weight gain and insulin resistance. Interaction between potential effects of fetal Zn restriction and food availability postnatally may be one factor responsible for later metabolic derangements. METHODS: Rats were fed Zn restricted (ZnD, 7 µg/g) or control (ZnC, 25 µg/g) diets ad libitum from 3 wk pre-conception to 3 wk post-parturition. Postnatally, litters were culled to 13 (IN, inadequate nutrition), 7 (AN, adequate nutrition), and 4 (EN, excess nutrition) pups/dam, respectively, and nursed by their original mothers. Postweaning, pups were fed rodent diet ad libitum. Tests to assess insulin resistance were performed subsequently.

Maternal zinc deficiency in rats affects growth and glucose metabolism in the offspring by inducing insulin resistance postnatally.

Interactions among zinc (Zn), insulin, and glucose metabolism are complex. Maternal Zn deficiency affects maternal carbohydrate metabolism, but the mechanisms underlying changes in glucose homeostasis of offspring are not well understood. Rats consumed Zn-deficient (ZnD; 7 microg/g) or control (ZnC; 25 microg/g) diets ad libitum from 3 wk preconception to 21 d postparturition. Litters were culled to 7 pups/dam postnatally and pups were allowed to nurse their original mothers; after weaning, pups were fed nonpurified diet. Insulin and glucose tolerance tests were performed on the pups at wk 5 and 10. Although there was no difference in birth weight between groups, ZnD pups weighed significantly more than controls by d 10 (+5%) and 20 (+10%). Both blood glucose and serum insulin-like growth factor (IGF-1) concentrations at wk 3 were significantly higher in ZnD pups than in controls. Both male and female ZnD rats were less sensitive to insulin and glucose stimulation than controls at wk 5 and 10. At wk 15, serum leptin concentrations were higher in male ZnD rats than in controls. Phosphorylation of muscle Akt protein, an insulin receptor (IR) signaling intermediate, was lower in female ZnD rats than in controls at wk 15, but they did not differ in phosphorylation of IR. Maternal Zn deficiency resulted in greater serum IGF-1 concentrations and the excessive postnatal weight gain in their offspring as well as impaired subsequent glucose sensitivity. It was associated with gender-specific alterations in the serum leptin concentration and the insulin signaling pathway. These findings suggest that suboptimal maternal Zn status induces long-term changes in the offspring related to abnormal glucose tolerance.

Effects of zinc exposure on zinc transporter expression in human intestinal cells of varying maturity.

OBJECTIVES: Zinc (Zn) homeostasis in adults is achieved principally through a balance between intestinal absorption and excretion involving adaptive mechanisms programmed by levels of dietary Zn. Zn absorption in infants is not as tightly regulated as that in adults, which may induce potential toxicity in infants due to the relatively high capacity of Zn absorption. We hypothesized that intestinal Zn homeostasis is developmentally regulated and depends on intestinal maturation, which in turn affects Zn transporter regulation. MATERIALS AND METHODS: Cultured human fetal (FHs 74 Int, F) and adult (Caco-2: undifferentiated, U; differentiated, D) intestinal cells were used to determine developmental differences in Zn uptake and effects of Zn exposure on Zn transporters. RESULTS: Zn uptake rates in F and U cells were higher compared with D cells (F, 9-fold; U, 3-fold). F cells were more intolerant to Zn exposure than were U or D cells (LD50 = 67.9 +/- 5.3; 117.0 +/- 5.2; 224.4 +/- 3.7 micromol/L, respectively). Two mechanisms were involved in developmental regulation of Zn homeostasis: differential Zn transporter expression and differential response to Zn exposure. In F cells, zinc-regulated transporter (ZRT)/iron-regulated transporter (IRT)-like protein (Zip)4 expression was undetectable; Zn (50 micromol/L) increased levels of Zn transporter (ZnT)1, ZnT2, and metallothionein-1 mRNA and ZnT1 protein. U and D cells had higher mRNA expression of ZnT1 (U: 5-fold; D: 7-fold, respectively) and ZnT2 (U: 2-fold; D: 9-fold, respectively) than F cells, and D cells also had higher Zip4 expression (3-fold) than U cells. In U cells, Zn exposure increased Zip4 protein level, but not membrane-associated abundance. However, in D cells, Zn exposure decreased both the Zip4 protein level and membrane-associated abundance. CONCLUSIONS: Zn absorption is developmentally regulated through intestinal Zn efflux and sequestration and import mechanisms, which may be responsible for differences in Zn absorption observed between infants and adults.

Tissue-specific alterations in zinc transporter expression in intestine and liver reflect a threshold for homeostatic compensation during dietary zinc deficiency in weanling rats.

Intestinal zinc (Zn) absorption and liver Zn mobilization are presumed to regulate Zn homeostasis. Several Zn transporters have been identified; however, their contribution to Zn homeostasis is poorly understood. Moreover, their regulation during periods of growth is unknown. To characterize the mechanisms that maintain Zn status, weanling rats were fed control (25 mg/kg), marginally low (MLZ; 15 mg/kg), low (LZ; 7 mg/kg), or very low (VLZ; <1 mg/kg) Zn diets for 3 wk and effects on jejunum Zip4 and ZnT1 and hepatic Zip1 and ZnT1 were assessed. Another control group was pair-fed (PF) to VLZ. The MLZ rats had lower jejunum ZnT1 protein abundance than the control. In the LZ group, we detected increased jejunum Zip4 mRNA expression and hepatic ZnT1 protein abundance and reduced jejunum Zip4 and ZnT1 and hepatic Zip1 protein abundance. VLZ had lower jejunum ZnT1 mRNA and protein abundance and hepatic Zip1 and ZnT1 protein abundance compared with the PF group. Zip4 protein was present at the intestinal villus tip in controls but was detected on the apical membrane throughout the entire villus in LZ rats. ZnT5 protein in jejunum was always detected at the apical membrane and also at the basolateral membrane of VLZ rats. In contrast, ZnT7 was found intracellularly in jejunum. Our data suggest that effects of Zn deficiency on Zn homeostasis occurs biphasically during marginal Zn deficiency through increased intestinal Zn uptake capacity and reduced intestinal Zn efflux, then during more pronounced degrees of Zn deficiency through decreased liver Zn accretion and increased hepatic Zn efflux back into circulation. These results assist in our understanding of how mammals regulate Zn homeostasis.

Insulin-like growth factor-I stimulates erythropoiesis when administered enterally.

BACKGROUND: Insulin-like growth factors I and II (IGF-I and IGF-II) are potent growth factors involved in development. IGF-I stimulates proliferation of erythropoietic progenitors and parenteral IGF-I administration stimulates in vivo erythropoiesis in animals. IGF-I and IGF-II are both present in mammalian milks and when milk-borne, are resistant to neonatal gastrointestinal degradation. Whether milk-borne IGF-I or IGF-II regulates neonatal erythropoiesis in not known. We hypothesized that physiological doses of enteral IGFs stimulate erythropoiesis in suckling rats. METHODS: Eight day-old Sprague Dawley rats were artificially fed for 4 days with rat milk substitute (RMS) or RMS supplemented with physiological levels of IGF-I or IGF-II. Rats fed IGF-I and IGF-II were compared to control RMS. Blood and marrow were collected; measures of red cell mass, measures of erythropoietic stimulus, and indices of iron status were measured. RESULTS: Rats fed IGF-I had higher hemoglobin (Hb) levels (100 +/- 10 g/l), compared to those fed RMS (94 +/- 9) or IGF-II (91 +/- 6), p < 0.001. After IGF-I supplementation, red blood cell counts (RBC) (p < 0.04) and hematocrits (p < 0.002) were also higher. Plasma erythropoietin (Epo) levels, reticulocytes, plasma iron and erythrocyte iron incorporation were similar. CONCLUSION: Intact enteral IGF-I reaches distal erythropoietic tissue resulting in greater red cell mass, but not by increasing plasma Epo levels or by altering cellular iron transport.

A graded model of dietary zinc deficiency: effects on growth, insulin-like growth factor-I, and the glucose/insulin axis in weanling rats.

OBJECTIVE: Severe zinc (Zn) deficiency inhibits growth, insulin storage and release. Mild or moderate Zn deficiency may also have profound physiological effects that are not outwardly evident. We examined the effects of graded levels of low Zn intake on growth, insulin-like growth factor-I (IGF-I) and glucose homeostasis in weanling rats. METHODS: Weanling rats were fed ad libitum for 3 weeks with diets containing different Zn levels: very low Zn, low Zn or mildly low Zn; there was also a control group and an additional group was pair-fed to very low Zn rats. Growth and food intake were recorded. Serum Zn, IGF-I, IGF binding protein-3 (IGFBP-3), serum insulin and glucose, tissue Zn and jejunal sucrase activity were measured. Relative liver IGF-I and IGFBP-3 mRNA levels were quantified. RESULTS: Serum and tissue Zn were significantly lower in rats fed very low Zn (compared with pair-fed animals and controls) and low Zn (compared with controls). Growth was significantly lower in rats fed very low Zn and pair-fed animals (compared with controls) and in those fed very low Zn (compared with pair-fed animals). Liver IGF-I and IGFBP-3 mRNA levels were higher in low Zn animals compared with controls. Serum IGF-1 and IGFBP-3 levels were not affected by diet. Serum glucose was significantly higher in rats fed very low Zn than in pair-fed animals (191 +/- 28 vs 99 +/- 5 mg/dL, respectively). Sucrase activity was lower in rats fed very low Zn than in pair-fed animals or controls and a linear relationship was observed between serum glucose and insulin (r = 0.65, P < 0.01) in pair-fed animals and controls but not in Zn-deficient groups. CONCLUSION: Severe Zn deficiency was associated with hyperglycemia and relative hypoinsulinemia. Mild degrees of Zn deficiency also altered glucose metabolism, suggesting that Zn intake may be a sensitive regulator of glucose homeostasis.

IGF alters jejunal glucose transporter expression and serum glucose levels in immature rats.

Milk-borne insulin-like growth factors (IGFs) enhance nutrient absorption in the immature intestine, which is characterized by low levels of glucose oxidation. We therefore hypothesized that feeding a rat milk substitute (RMS) devoid of growth factors to rat pups would lower serum glucose levels relative to dam-fed control rats and that supplementation of RMS with physiological doses of either IGF-I or IGF-II would normalize serum glucose levels via increased jejunal glucose transporter 2 (GLUT2) and high-affinity Na(+)-glucose cotransporter (SGLT1) expression. We found lower serum glucose concentrations in RMS-fed pups; in contrast, serum glucose levels in the IGF-supplemented pups were similar to those of dam-fed controls. RT-PCR and laser scanning confocal microscopy similarly demonstrated that IGF supplementation increased expression of jejunal glucose transporters. Further experiments demonstrated that IGF supplementation altered mRNA levels of key mitochondrial enzymes without altering jejunal lactase activity. We conclude that IGF-I and IGF-II supplementation increases serum glucose levels in the immature rat pup fed artificial formula and alters gene expression of the jejunal glucose transporters.

Intestinal transport of insulin-like growth factor-I (igf-I) in the suckling rat.

OBJECTIVES: Insulin-Like Growth Factor-1 is a potent growth-promoting peptide that is present in mammalian milk. Previous studies have suggested that milk-borne IGF-1 may be absorbed intact from the gastrointestinal tract of the suckling but the mechanism responsible for such transport is not well documented. The present study was designed to investigate in an in vivo suckling rat model whether or not intestinal absorption of IGF-1 is a saturable phenomenon. METHODS: Suckling rats (10-12 days postnatal age) were studied under anesthesia. A jejunal loop from each rat pup was isolated and injected intraluminally with 1-2 x 10 cpm of rh I-IGF-I. Injections were performed in paired littermates either with or without a preceding injection of unlabeled IGF-I of 20, 500, or 1000 ng/ml concentration. After flushing, the loops and livers were homogenized and counted in a gamma counter. In addition, homogenates of jejunum and liver were precipitated with trichloroacetic acid (TCA) and the precipitates also counted. In selected instances (jejunum), acid gel chromatography of homogenates was also performed. RESULTS: Retention of radioactivity was observed in all jejunal specimens, but the pre-incubation of jejunal loops with unlabeled IGF-1 was associated with a biphasic response, i.e. at low dose (20 ng/ml) pre-incubation limited retention of radioactivity, but at a high dose (1000 ng/ml), retention was enhanced (P < 0.05). Linear regression analysis confirmed this inverse relationship. Liver radioactivity followed a similar pattern. Between 40 and 49% of the radioactivity in jejunal and liver homogenates was TCA precipitable. Chromatography of jejunal homogenates showed that approximately 40% of cpm migrated in a position identical with that of intact IGF-1. CONCLUSIONS: The intestinal uptake of IGF-1 in the suckling is nonsaturable, confirming previous in vitro studies and suggesting that a nonreceptor-dependent method of transepithelial transport is important in this process.