Iron-adequate Diet Research Articles

Classical Selenium-Dependent Glutathione Peroxidase Expression Is Decreased Secondary to Iron Deficiency in Rats

While there are reports that classical selenium-dependent glutathione peroxidase (Se-GPX1) activity is decreased during iron deficiency, the relationship between tissue iron status and Se-GPX1 activity remains speculative. This study was undertaken to investigate the mechanism for the decrease in Se-GPX1 activity during iron deficiency. Male weanling Sprague-Dawley rats were given free access to either an iron-deficient or an iron-adequate diet for eight weeks, after which blood, livers, kidneys, hearts, brains and testes were surgically excised. During iron deficiency, Se-GPX1 mRNA levels in liver tissue were decreased by approximately 55%. Similarly, the concentration of immunoreactive Se-GPX1 protein and total selenium-dependent glutathione peroxidase (Se-GPX) activity were decreased by 55% and 60%, respectively. In kidney, heart and brain total Se-GPX activities were depressed as much as 33%. Selenium concentration in liver was reduced by 42%, whereas the decrease in Se concentrations in kidney, heart, and brain ranged from 17 to 25%. Concentrations of plasma Se also were reduced by 18%, but testes showed little change in either Se-GPX activity or Se concentration during iron deficiency. Results suggest that the synthesis of Se-GPX1 protein is decreased during iron deficiency possibly due to pretranslational regulation.

Pectin with Low Molecular Weight and High Degree of Esterification Increases Absorption of 58Fe in Growing Rats

Effects of pectins with different degrees of esterification (DE) and molecular weights (MW) on iron bioavailability were investigated in healthy growing rats by following erythrocyte incorporation of a dose of 58Fe. Rats were fed a control diet for 8 d and then deprived of food for 16 h. Two hours after the start of feeding iron-deficient diets, with or without pectin (80 g/kg diet), a dose of FeSO4 rich in 58Fe (60.28%) was intubated into the stomach; rats were then allowed to feed for an additional 4 h before withdrawal of food for 10 h. Rats were then fed iron-adequate diets for 9 d. The pectins differed in DE and MW, respectively, as follows: P-A (73%, 860,000), P-B (75%, 89,000), P-C (22%, 1,260,000) and P-D (24%, 114,000). Rats fed pectin-free diet with free access to food or restricted to the same quantity consumed by a respective pectin group served as controls. Iron absorption was 48% in the control group and 57% in rats fed P-B. Rats fed P-B had higher (P ≤ 0.05) serum iron, transferrin saturation, hematocrit and liver and spleen iron than the control group or the group fed P-C. These indices, except for transferrin saturation, were also higher in rats fed P-A and P-D compared with those fed P-C and controls, but to a lesser extent than in rats fed P-B. The data indicate that bioavailability of dietary non-heme iron was enhanced when pectin of low MW and high DE was added to the diet. This improvement was not evident with pectins having high MW and/or low DE.

Gestational Iron Supplementation Improves Fetal Outcomes in a Rat Model of Prenatal Alcohol Exposure.

Prenatal alcohol exposure causes neurodevelopmental disability and is associated with a functional iron deficiency in the fetus and neonate, even when the mother consumes an apparently iron-adequate diet. Here, we test whether gestational administration of the clinically relevant iron supplement Fer-In-Sol mitigates alcohol’s adverse impacts upon the fetus. Pregnant Long-Evans rats consumed an iron-adequate diet and received 5 g/kg alcohol by gavage for 7 days in late pregnancy. Concurrently, some mothers received 6 mg/kg oral iron. We measured maternal and fetal weights, hematology, tissue iron content, and oxidative damage on gestational day 20.5. Alcohol caused fetal anemia, decreased fetal body and brain weight, increased hepatic iron content, and modestly elevated hepatic malondialdehyde (p’s < 0.05). Supplemental iron normalized this brain weight reduction in alcohol-exposed males (p = 0.154) but not female littermates (p = 0.031). Iron also reversed the alcohol-induced fetal anemia and normalized both red blood cell numbers and hematocrit (p’s < 0.05). Iron had minimal adverse effects on the mother or fetus. These data show that gestational iron supplementation improves select fetal outcomes in prenatal alcohol exposure (PAE) including brain weight and hematology, suggesting that this may be a clinically feasible approach to improve prenatal iron status and fetal outcomes in alcohol-exposed pregnancies.

Baseline iron status and presence of anaemia determine the course of systemic Salmonella infection following oral iron supplementation in mice

Iron deficiency anaemia (IDA) is a major health concern. However, preventive iron supplementation in regions with high burden of infectious diseases resulted in an increase of infection related morbidity and mortality. We fed male C57BL/6N mice with either an iron deficient or an iron adequate diet. Next, they received oral iron supplementation or placebo followed by intraperitoneal infection with Salmonella Typhimurium (S.Tm). We found that mice with IDA had a poorer clinical outcome than mice on an iron adequate diet. Interestingly, iron supplementation of IDA mice resulted in higher bacterial burden in organs and shortened survival. Increased transferrin saturation and non-transferrin bound iron in the circulation together with low expression of ferroportin facilitated the access of the pathogen to iron and promoted bacterial growth. Anaemia, independent of iron supplementation, was correlated with reduced neutrophil counts and cytotoxic T cells. With iron supplementation, anaemia additionally correlated with increased splenic levels of the cytokine IL-10, which is suggestive for a weakened immune control to S.Tm infection. Supplementing iron to anaemic mice worsens the clinical course of bacterial infection. This can be traced back to increased iron delivery to bacteria along with an impaired anti-microbial immune response. Our findings may have important implications for iron supplementation strategies in areas with high endemic burden of infections, putting those individuals, who potentially profit most from iron supplementation for anaemia, at the highest risk for infections. Financial support by the Christian Doppler Laboratory for Iron Metabolism and Anemia Research.

Dietary Iron Repletion Stimulates Hepatic Mobilization of Vitamin A in Previously Iron-Deficient Rats as Determined by Model-Based Compartmental Analysis

Iron deficiency can result in hyporetinolemia and hepatic vitamin A (VA) sequestration. We used model-based compartmental analysis to determine the impact of iron repletion on VA metabolism and kinetics in iron-deficient rats. At weaning, Sprague-Dawley rats were assigned to either a VA-marginal diet (0.35mg retinol equivalent/kg) with adequate iron (35ppm, control group [CN]) or reduced iron (3ppm, iron-deficient group [ID-]), with an equivalent average body weight for each group. After 5 wk, n=4 rats from each group were euthanized for baseline measurements of VA and iron indices, and the remaining rats (n=6 CN, n=10 ID-) received an intravenous injection of 3H-labeled retinol in an emulsion as tracer to initiate the kinetic study. On day 21 after dosing, half of the ID- rats were switched to the CN diet to initiate iron repletion, referred to as the iron-repletion group (ID+). From the time of dosing, 34 serial blood samples were collected from each rat over a 92-d time course. Plasma tracer and tissue tracee data were fitted to 6- and 4-compartment models, respectively, to analyze the kinetic behavior of VA in all groups. Our mathematical model indicated that ID- rats exhibited a nearly 6-fold decrease in liver VA secretion and >4-fold reduction in whole-body VA utilization, compared with CN rats, whereas these perturbed kinetic behaviors were notably corrected in ID+rats, close to those from the CN group. Iron repletion can remove the inhibitory effect that iron deficiency exerts on hepatic mobilization of VA and restore retinol kinetic parameters to values similar to that of never-deficient CN rats. Together with improvements in iron and VA indices, our results suggest that restoration of an iron-adequate diet is sufficient to improve VA kinetics after a previous state of iron deficiency.

The Clinical Iron Supplement Fer-In-Sol Improves Fetal Iron Status and Brain Weight in Rats Prenatally Exposed to Alcohol

Abstract Objectives Prenatal alcohol exposure (PAE) causes iron deficiency in fetal rats, even when their mothers consume enough iron. Increasing maternal dietary iron improves fetal outcomes, including blood status, brain iron, and proinflammatory cytokines. Clinically, gestational iron deficiency is widespread, with a great need for effective iron interventions in alcohol-exposed pregnancies. Here, we test the ability of a popular clinical iron supplement, Fer-In-Sol (ferrous sulfate), to mitigate PAE's effects when co-administered with alcohol during pregnancy. We hypothesized that the prenatal iron supplement would normalize fetal iron status and have minimal adverse effects on the PAE dams and fetuses. Methods Pregnant Long-Evans rats consumed an iron-adequate diet (AIN-76A, 100 ppm iron) and received 5 g/kg alcohol or isocaloric maltodextrin (MD) by gavage on gestational days (GD) 13.5–19.5. Some dams received 6 mg/kg oral iron as ferrous sulfate from GD12.5–19.5, generating a 2 × 2 design. At GD20.5, we used an ANOVA to analyze mothers and fetuses, with litter as the experimental unit. Results Iron had no effect on PAE-induced reductions in maternal gestational weight gain (P = 0.7818) or maternal food intake (P = 0.7299). Iron did not mitigate a PAE-induced 17% reduction in fetal weight (P = 0.4803). Although iron failed to improve a 10% reduction in placental efficiency (P = 0.0589) or 10% elevation in relative heart weight (P = 0.2958) due to PAE, iron did elevate PAE fetal brain weight by 5%, and it no longer differed from MD-controls (P = 0.5255). Furthermore, iron normalized fetal hematology values in PAE, and improved fetal red blood cell numbers (P = 0.0458) and hematocrit (P = 0.0439) and trended to normalize hemoglobin (P = 0.0585). Iron did not significantly alter maternal blood, suggesting that this iron dose was not excessive. As expected, PAE increased malondialdehyde (MDA) levels in maternal (+10%, P = 0.0224) and fetal livers (+44%, P &amp;lt; 0.0001). Iron supplementation caused an additional, modest rise in maternal (+15%, P = 0.0042) and fetal (+22%, P = 0.0093) livers. Conclusions These data show that maternal iron supplementation improves select fetal outcomes in PAE, including brain weight and blood values, and suggests that this may be a clinically feasible approach to improve prenatal iron status and fetal outcomes in PAE pregnancies. Funding Sources NIAAA NIDDK.

Postweaning Iron Deficiency in Male Rats Leads to Long-Term Hyperactivity and Decreased Reelin Gene Expression in the Nucleus Accumbens

Background:Epidemiological research indicates that iron deficiency (ID) in infancy correlates with long-term cognitive impairment and behavioral disturbances, despite therapy. However, the mechanisms underlying these effects are unknown. Objectives:We investigated how ID affected postweaning behavior and monoamine concentration in rat brains to determine whether ID during the juvenile period affected gene expression and synapse formation in the prefrontal cortex (PFC) and nucleus accumbens (NAcc). Methods:Fischer 344/Jcl postweaning male rats aged 21–39 d were fed low-iron diets (0.35 mg/kg iron; ID group) or standard AIN-93 G diets [3.5 mg/kg iron; control (CN) group]. After day 39, all rats were fed the iron-adequate diet. The locomotor activity was evaluated by the open field and elevated plus maze tests at 8 and 12 wk of age. Monoamine concentrations in the brain were analyzed using HPLC at 9 and 13 wk of age. Comprehensive gene expression analysis was performed in the PFC and NAcc at 13 wk of age. Finally, we investigated synaptic density in the PFC and NAcc by synaptophysin immunostaining. Results:Behavioral tests revealed a significant reduction of the age-related decline in the total distance traveled in ID rats compared with CN rats (P < 0.05), indicating that ID affected hyperactivity, which persisted into adulthood (13 wk ofage). At this age, reelin (Reln) mRNA expression (adjusted P < 0.01) decreased and synaptic density (P < 0.01) increasedin the NAcc in the ID group. Regarding the mesolimbic pathway, homovanillic acid concentration increased in the NAcc,whereas the dopamine concentration decreased in the ventral midbrain. Conclusions:Our results suggest that ID during the postweaning period in male rats, despite complete iron repletion following ID, led to long-term hyperactivity via monoamine disturbance in the brain and an alteration in the synaptic plasticity accompanied by downregulation of Reln expression in the NAcc.

Early-Life Iron Deficiency and Subsequent Repletion Alters Development of the Colonic Microbiota in the Pig.

Background: Iron deficiency is the most prevalent micronutrient deficiency worldwide, affecting over two billion people. Early-life iron deficiency may alter the developing microbiota, which may or may not be reversible with subsequent dietary iron repletion. Thus, the aim of this study was to determine whether early-life iron deficiency and subsequent repletion alter colonic microbial composition and fermentation end-product concentrations in pigs.Methods: Forty-two male pigs received either control (CONT, 21.3 mg Fe/L) or iron-deficient (ID, 2.72 mg Fe/L) milk replacer treatments from postnatal day (PND) 2 to 32. Subsequently, 20 pigs continued through a series of age-appropriate, iron-adequate diets from PND 33 to 61. Contents from the ascending colon (AC) and rectum (feces) were collected at PND 32 and/or 61. Assessments included microbiota composition by 16S rRNA sequencing and volatile fatty acid (VFA) concentrations by gas chromatography methods. Data were analyzed using a 1-way ANOVA and PERMANOVA to assess the main effects of early-life iron status on all outcomes.Results: In AC samples, 15 genera differed (P < 0.05) between ID and CONT pigs, while 27 genera differed (P < 0.05) in fecal samples at PND 32. Early-life ID pigs had higher (P = 0.012) relative abundance of Lactobacillus in AC samples compared with CONT pigs. In feces, ID pigs had lower (P < 0.05) relative abundances of Bacteroides and Clostridium from the families of Clostridiaceae, Lachnospiraceae, and Ruminococcaceae. At PND 61, only two genera differed between treatment groups in AC samples, with ID pigs having a higher (P = 0.043) relative abundance of Bifidobacterium and lower (P = 0.047) relative abundance of Prevotella. Beta diversity differed at PND 32 in both AC and feces between CONT and ID pigs but no differences remained at PND 61. At PND 32, the total VFA concentration was higher in ID pigs compared with CONT pigs in both AC (P = 0.003) and feces (P = 0.001), but no differences in VFA concentrations persisted to PND 61.Conclusion: Early-life iron status influenced microbial composition and VFA concentrations within the large intestine, but these differences were largely normalized following subsequent dietary iron repletion.

Longitudinal Effects of Iron Deficiency Anemia and Subsequent Repletion on Blood Parameters and the Rate and Composition of Growth in Pigs.

Iron deficiency is reported as the most common nutrient deficiency worldwide. Due to rapid growth, infants are at particular risk for developing iron deficiency, which can easily progress to iron deficiency anemia (IDA), if not treated. The aim of this study was to determine the lasting effects of an early-life iron deficiency after a period of dietary iron repletion. Forty-two intact male pigs were fed, ad libitum, either control (CONT, 21.3 mg Fe/L) or iron-deficient (ID 2.72 mg Fe/L) milk replacer from postnatal day (PND) 2 to 32 (phase 1). From PND 33 to 61 (phase 2), all pigs were transitioned onto a series of industry-standard, iron-adequate diets. Blood was collected weekly from PND 7 to 28, and again on PND 35 and 56, and tissues were collected at either PND 32 or PND 61. At the end of phase 1, ID pigs exhibited reduced hematocrit (Hct; p < 0.0001) and hemoglobin (Hb; p < 0.0001) compared with CONT pigs, but neither Hct (p = 0.5968) nor Hb (p = 0.6291) differed between treatment groups after dietary iron repletion at the end of phase 2. Body weight gain was reduced (p < 0.0001) 58% at PND 32 in ID pigs compared with CONT pigs during phase 1, and this effect remained significant at the end of phase 2 (p = 0.0001), with ID pigs weighing 34% less than CONT pigs at PND 61. Analysis of peripheral protein and messenger RNA (mRNA) gene expression biomarkers yielded inconclusive results, as would be expected based on previous biomarker analyses across multiple species. These findings suggest that early-life iron status negatively influences blood parameters and growth performance, with dietary iron repletion allowing for full recovery of hematological outcomes, but not growth performance.

Severe iron deficiency and manganese concentrations in the rat cerebrum

Severe iron deficiency is known to increase brain manganese concentration in rats. We studied whether severe iron deficiency affects dose‐effect relationship between dietary manganese and brain manganese in rats. Forty‐eight weaning male Wistar rats were equally divided into 8 groups and fed on respective experimental diets for 7 weeks. Four diets were iron‐deficient diet containing graded levels of manganese: one times, 3 times, 6 times and 9 times the National Research Council manganese requirement. The other four diets were iron‐adequate diet containing the graded levels of manganese as same as the above. After dietary regimen and 16 hour starvation, rat cerebrum was separated into striate, hippocampus and cortex (i.e., remaining parts). Samples were decomposed by wet ashing using nitric acid followed by hydrogen peroxide. Manganese concentration was measured by inductively coupled plasma‐mass spectrometry. Data were analyzed by simple linear regression, general linear model and nonlinear regression. The model that gave the highest Akaike information criterion was selected. Except for hippocampal manganese concentration in iron‐adequate rats, manganese concentrations in the measured brain regions were significantly and positively correlated with dietary manganese level. For hippocampus and striate, a general linear model was selected. Iron deficiency and dietary manganese independently and significantly increased manganese level in striate. In hippocampus, iron deficiency significantly increased manganese concentration. Effect of dietary manganese was significant when animals were iron‐deficient. Exponential saturation model regarding dietary manganese was selected for cortex. Effect of dietary manganese was significant and iron deficiency increased the curve span approximately by two‐fold. These results suggest that severe iron deficiency augments increments of manganese level by dietary manganese in the rat brain region.Support or Funding InformationThis work was supported by JSPS KAKENHI Grant Numbers JP17K00877 and JP 25350137.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Regulation of trace elements and redox status in striatum of adult rats by long-term aerobic exercise depends on iron uptakes

We investigated the effects of aerobic exercise (AE) on trace element contents and redox status in the striatum of rats with different diet iron. Weaned female rats were randomly fed with iron-adequate diet (IAD), iron-deficient diet (IDD), and iron-overloaded diet (IOD). After feeding their respective diet for 1 month, the rats fed with same diet were divided into swimming and maintaining sedentary (S) group. After 3 months, the non-heme iron (NHI), Mn, Cu, and Zn in the striatum were measured. Meanwhile, malonaldehyde acid (MDA), total superoxide dismutase activity, hydroxyl radical scavenging activity, and total antioxidant capacity were also analyzed. As compared with respective S rats, Mn, Cu, and Zn contents were significantly decreased in IDDE, but no significantly changes could be seen in IADE or IODE. A negative correlation of NHI with Cu contents in IDDE and positive correlations of NHI with Cu, or Zn contents in IADE, or with Mn or Cu contents in IODE were observed. In addition, striatum MDA was significantly decreased and anti-oxidative variables were increased in IODE compared to IODS. Our results suggest that the modification of trace elements and redox status in the striatum of rats caused by AE depends on dietary iron contents and that AE may also regulate the metabolic relationship of iron storage with other trace elements.

Synergistic Inhibitory Effects of Hypoxia and Iron Deficiency on Hepatic Glucose Response in Mouse Liver.

Hypoxia and iron both regulate metabolism through multiple mechanisms, including hypoxia-inducible transcription factors. The hypoxic effects on glucose disposal and glycolysis are well established, but less is known about the effects of hypoxia and iron deficiency on hepatic gluconeogenesis. We therefore assessed their effects on hepatic glucose production in mice. Weanling C57BL/6 male mice were fed an iron-deficient (4 ppm) or iron-adequate (35 ppm) diet for 14 weeks and were continued in normoxia or exposed to hypoxia (8% O2) for the last 4 weeks of that period. Hypoxic mice became hypoglycemic and displayed impaired hepatic glucose production after a pyruvate challenge, an effect accentuated by an iron-deficient diet. Stabilization of hypoxia-inducible factors under hypoxia resulted in most glucose being converted into lactate and not oxidized. Hepatic pyruvate concentrations were lower in hypoxic mice. The decreased hepatic pyruvate levels were not caused by increased utilization but rather were contributed to by decreased metabolism from gluconeogenic amino acids. Pyruvate carboxylase, which catalyzes the first step of gluconeogenesis, was also downregulated by hypoxia with iron deficiency. Hypoxia, and more so hypoxia with iron deficiency, results in hypoglycemia due to decreased levels of hepatic pyruvate and decreased pyruvate utilization for gluconeogenesis. These data highlight the role of iron levels as an important determinant of glucose metabolism in hypoxia.

Neurobehavioural analysis of developmental iron deficiency in Oreochromis aureus × Oreochromis niloticus.

The objective of this study was to examine the association between brain iron measurements of monoamine function and behavioural measurements of learning and memory. Male hybrid tilapias Oreochromis aureus × Oreochromis niloticus were fed either an iron-deficient (ID) diet or an iron-adequate (IA) diet for 8 weeks. The ID fishes showed significantly lower iron content in brain and decreasing learning and memory capacity. The fishes that showed increased learning and memory capacity had higher levels of iron and monoamine oxidase activity in brain. In addition, the results showed that learning and memory behaviours were related to monoamine (dopamine and noradrenaline) concentration in the brain. This suggests that iron can enhance learning and memory capacity in fishes and that the effect may have monoaminergic mediation in discrimination learning and memory tasks. The experimental data suggest that the properties and neural basis of learning and memory of teleosts are notably similar to those of land vertebrates.

Iron deficiency alters expression of dopamine-related genes in the ventral midbrain in mice

A clear link exists between iron deficiency (ID) and nigrostriatal dopamine malfunction. This link appears to play an important role in at least restless legs syndrome (RLS) if not several other neurological diseases. Yet, the underlying mechanisms remain unclear. The effects of ID on gene expression in the brain have not been studied extensively. Here, to better understand how exactly ID alters dopamine functioning, we investigated the effects of ID on gene expression in the brain, seeking to identify any potential transcription-based mechanisms. We used six strains of recombinant inbred mice (BXD type) known to differ in susceptibility to ID in the brain. Upon weaning, we subjected mice from each strain to either an iron-deficient or iron-adequate diet. After 100days of dietary treatment, we measured the effects of ID on gene expression in the ventral midbrain, a region containing the substantia nigra. The substantia nigra is the base of the nigrostriatal dopamine pathway and a region particularly affected by iron loss in RLS. We screened for ID-induced changes in expression, including changes in that of both iron-regulating and dopamine-related genes. Results revealed a number of expression changes occurring in ID, with large strain-dependent differences in the genes involved and number of expression changes occurring. In terms of dopamine-related genes, results revealed ID-induced expression changes in three genes with direct ties to nigrostriatal dopamine functioning, two of which have never before been implicated in an iron–dopamine pathway. These were stromal cell-derived factor 1 (Cxcl12, or SDF-1), a ferritin regulator and potent dopamine neuromodulator, and hemoglobin, beta adult chain 1 (Hbb-b1), a gene recently shown to play a functional role in dopaminergic neurons. The extent of up-regulation of these genes varied by strain. This work not only demonstrates a wide genetic variation in the transcriptional response to ID in the brain, but also reveals two novel biochemical pathways by which iron may potentially alter dopamine function.

Systems genetic analysis of multivariate response to iron deficiency in mice

The aim of this study was to identify genes that influence iron regulation under varying dietary iron availability. Male and female mice from 20+ BXD recombinant inbred strains were fed iron-poor or iron-adequate diets from weaning until 4 mo of age. At death, the spleen, liver, and blood were harvested for the measurement of hemoglobin, hematocrit, total iron binding capacity, transferrin saturation, and liver, spleen and plasma iron concentration. For each measure and diet, we found large, strain-related variability. A principal-components analysis (PCA) was performed on the strain means for the seven parameters under each dietary condition for each sex, followed by quantitative trait loci (QTL) analysis on the factors. Compared with the iron-adequate diet, iron deficiency altered the factor structure of the principal components. QTL analysis, combined with PosMed (a candidate gene searching system) published gene expression data and literature citations, identified seven candidate genes, Ptprd, Mdm1, Picalm, lip1, Tcerg1, Skp2, and Frzb based on PCA factor, diet, and sex. Expression of each of these is cis-regulated, significantly correlated with the corresponding PCA factor, and previously reported to regulate iron, directly or indirectly. We propose that polymorphisms in multiple genes underlie individual differences in iron regulation, especially in response to dietary iron challenge. This research shows that iron management is a highly complex trait, influenced by multiple genes. Systems genetics analysis of iron homeostasis holds promise for developing new methods for prevention and treatment of iron deficiency anemia and related diseases.

A Postweaning Iron-Adequate Diet Following Neonatal Iron Deficiency Affects Iron Homeostasis and Growth in Young Rats ,

Iron deficiency is among the most prevalent of nutrient-related diseases worldwide, but the long-term consequences of maternal and neonatal iron deficiency on offspring are not well characterized. We investigated the effects of a postweaning iron-adequate diet following neonatal iron deficiency on the expression of genes involved in iron acquisition and homeostasis. Pregnant rats were fed an iron-adequate diet (0.08 g iron/kg diet) until gestational d 15, at which time they were divided into 2 groups: 1) a control group fed an iron-adequate diet, and 2) an iron-deficient group fed an iron-deficient diet (0.005 g iron/kg diet) through postnatal d (P) 23 (weaning). After weaning, pups from both dietary treatment groups were fed an iron-adequate diet until adulthood (P75). Rat pups that were iron deficient during the neonatal period (IDIA) had reduced weight gain and hemoglobin concentrations and decreased levels of serum, liver, and spleen iron on P75 compared with rats that were iron sufficient throughout early life (IA). IDIA rats developed erythrocytosis during postweaning development. Further, hepatic expression of hepcidin in IDIA rats was 1.4-fold greater than in IA rats, which paralleled an upregulation of IL-1 expression in the serum. Our data suggest that an iron-adequate diet following neonatal iron deficiency induced an inflammatory milieu that affected iron homeostasis and early growth and development.

Interrelationships between tissue iron status and erythropoiesis during postweaning development following neonatal iron deficiency in rats

Dietary iron is particularly critical during periods of rapid growth such as in neonatal development. Human and rodent studies have indicated that iron deficiency or excess during this critical stage of development can have significant long- and short-term consequences. Since the requirement for iron changes during development, the availability of adequate iron is critical for the differentiation and maturation of individual organs participating in iron homeostasis. We have examined in rats the effects of dietary iron supplement following neonatal iron deficiency on tissue iron status in relation to erythropoietic ability during 16 wk of postweaning development. This physiological model indicates that postweaning iron-adequate diet following neonatal iron deficiency adversely affects erythroid differentiation in the bone marrow and promotes splenic erythropoiesis leading to splenomegaly and erythrocytosis. This altered physiology of iron homeostasis during postweaning development is also reflected in the inability to maintain liver and spleen iron concentrations and the altered expression of iron regulatory proteins in the liver. These studies provide critical insights into the consequences of neonatal iron deficiency and the dietary iron-induced cellular signals affecting iron homeostasis during early development.

Iron deficiency affects acoustic startle response and latency, but not prepulse inhibition in young adult rats

Iron deficiency is associated with alterations in dopamine and serotonin transporters as well as changes in dopamine receptor (DR) density, monoamine concentrations, and in vivo extracellular contents of monoamines in terminal fields. Human infants with iron deficiency have both delayed maturation as well as lengthened central conduction times in auditory evoked potential studies. The current study utilizes the magnitude of the acoustic startle response (ASR), prepulse inhibition (PPI), and mean latency to maximum startle response ( T max), to examine the functional integrity of response to environmental cues. Male and female rats consumed iron deficient (ID) or iron adequate (CN) diets from weaning until adulthood. ID rats of both sexes had 20–60% reductions in ASR when compared to CN rats but there was no effect on PPI. T max was significantly longer by 10–20% in females, but not males. Dopamine transporter density was significantly lower in putamen, nucleus accumbens, and olfactory tubercle in males, but not female rats while the serotonin transporter was significantly different from control animal density in five of 14 brain regions. Norepinephrine transporter density was lower in the locus ceruleus of ID male rats but was unaffected in ID female rats. Regression modeling of ASR with brain monoamine transporters and receptors showed hematocrit, norepinephrine transporter (NET) in dentate gyrus, and D 1R in the nucleus accumbens account for nearly 49% of the variance in ASR. T max was not significantly associated with any of the independent variables. We conclude that iron deficiency affects the startle response, but not the inhibitory circuits involved in prepulse inhibition. Importantly, sex also strongly influenced these behavioral responses. Future studies, perhaps pharmacologic in nature, are necessary to ascertain whether iron deficiency modifies the contribution of monoaminergic systems to responses to environmental stimuli.

Non‐haem iron transport in the rat proximal colon

Only 10% of dietary iron is absorbed in the duodenum which implies that 90% (approximately 9 mg day(-1)) reaches the lower small intestine and colon. Therefore the purpose of this study was to assess the iron transport capacity of the rat proximal colon and to determine whether iron absorption is regulated by changes in dietary iron content. Rats were fed for 14 days on either iron adequate (44 mg Fe kg(-1) diet) or iron-deficient (< 0.5 mg Fe kg(-1) diet) diets. The 59Fe transport across the colonic epithelium and its subsequent appearance in the blood were measured in vivo. In separate studies the colon was excised and used to measure divalent metal transporter expression. Divalent metal transporter (DMT1) was expressed at the apical membrane of the surface epithelium in rat proximal colon. In animals fed an iron-deficient diet, DMT1 mRNA and protein expression were increased. This was accompanied by a significant increase in tissue 59Fe uptake. The proximal colon can absorb non-haem iron from the intestinal lumen. The purpose of this mechanism remains to be elucidated.

Iron deficiency: Differential effects on monoamine transporters

In this study, we extend previous work on iron deficiency and dopamine (DA) transporters to include an examination of central serotonin (5-HT) and noradrenergic (NE) transporters. Rats were fed either iron deficient (ID) or iron adequate (CN) diets from weaning until adulthood. In males, an additional group of iron deficient animals (IR) were given iron supplementation. DA, 5-HT, and NE transporter binding was done in situ on thin sections. ID males, but not females, decreased DA transporter binding in the nucleus accumbens, caudate putamen and substantia nigra by 20–40%. ID males also had a 20–30% reduction in 5-HT transporter binding in several areas (nucleus accumbens, olfactory tubercle, colliculus) while in ID females there was 15–25% increased serotonin transporter binding in the olfactory tubercle, zona incerta, anteroventral thalamic nucleus and vestibular nucleus. Iron deficiency reduced 3 H-nisoxetine binding to the NE transporter in locus ceruleus and anteroventral thalamic nucleus in males but not females. Only some of the changes observed in DA, serotonin and NE transporter binding were reversible by iron supplementation. These findings show that iron deficiency affects monoamine systems related to homeostasis and in most cases males appear to be more vulnerable than females.