Zinc deficiency and behavior: A developmental perspective

Zinc deficiency was induced in 35 and 300 day-old male Holtzman rats. Group ZDA was fed ad lib a diet deficient in zinc (1 ppm), ZSP controls were pair-fed a diet supplemented with zinc (50 ppm) and ZSA controls were fed ad lib a diet supplemented with zinc (50 ppm). Physical status and six open-field behaviors were evaluated. Food intake, body weight and plasma zinc concentrations were significantly reduced in both age groups. Expanded use of the open-field revealed significantly lower latencies to explore the novel environment and significantly lower ambulation scores in the young and older zinc deficient rats. Older rats spent significantly less time grooming than their controls. Rearing was significantly less in young zinc deficient rats and “kangaroo-like” posture was evident. Young rats made deficient during critical periods of growth and development, were at greater risk for most parameters tested, compared to the older deficient rats. These results do demonstrate, however, that feeding low-zinc diets to older, fully developed animals results in significant physical and behavioral impairment.     

Learning and memory disabilities in young adult rats from mildly zinc deficient dams

Three experiments were conducted to test the behavior of the offspring of rat dams (ZD) fed a mildly zinc deficient diet (10.0 micrograms Zn/g) during pregnancy and lactation. Since zinc deficiency causes anorexia, a second group of rat dams (PF) was fed the same quantity of the diet during gestation and lactation as was consumed by their ZD mates. A third group of rats (AL) was fed the diet ad lib during gestation and lactation. The PF and AL dams were given zinc supplemented (25.0 micrograms Zn/ml) drinking water whereas the ZD dams were given double distilled, deionized drinking water. After weaning, the offspring of all dams were fed Purina Laboratory Chow ad lib until they were 100 days old. The offspring were then reduced to 85% of their ad lib weight and tested on a 17-arm radial maze for memory and learning. In Experiments 1 and 2, the ZD males suffered a significant learning deficit when compared to the AL males. Whereas the PF males suffered a significant learning impairment in Experiment 1, the learning deficit of the PF group was not as severe as the deficit of the ZD group. There was no impairment in reference (long-term) memory for any of the groups. In Experiment 3, significant differences in working (short-term) memory were found among the three groups of females. The ZD group was significantly inferior in working memory when compared to the PF and AL groups. No significant differences in working memory were found between the PF and AL groups.     

The role of the endogenous opiates in zinc deficiency anorexia

Anorexia is a major symptom of zinc deficiency, but the mechanism(s) for this anorexia are poorly defined. Recent studies have suggested an integral role for endogenous opiate peptides in appetite regulation. Dynorphin, a leucine-enkephalin containing opiate peptide, is a potent inducer of spontaneous feeding. In this study we showed that zinc deficient animals were relatively resistant to dynorphin-induced feeding. Measurement of dynorphin levels using a highly sensitive radioimmunoassay showed that zinc deficient animals had lower levels of dynorphin in the hypothalamus than did ad lib fed animals, with weight restricted animals having intermediate values. [3H]-naloxone binding was significantly increased to isolated brain membranes from zinc deficient animals using 1 nM unlabeled naloxone when compared to ad lib fed controls with the weight restricted animals again having intermediate values. These data suggest that abnormalities in endogenous opiate regulation of appetite may well play a role in the anorexia of zinc deficiency. The effects of zinc deficiency on endogenous opiate action appear to include alterations in receptor affinity, a post-receptor defect and alterations in the synthesis and/or release of dynorphin.     

Development of food motivated behavior in free feeding and food restricted zucker fatty (fa/fa) rats

The food motivated behavior of ad lib fed or calorically restricted male Zucker fatty and lean rats was compared at 12, 16, 20 and 30 weeks of age. The groups were fasted for 0, 12 and 24 hr and required to lever press for food pellets on VI 40 sec. Responding by ad lib fatties was elevated over lean controls during the rapid growth period (12 and 16 weeks of age) but was significantly reduced following the completion of growth (20 weeks). Elevated responding accompanied hyperphagia by the ad lib fatties during growth but did not accompany a second, adult onset period of hyperphagia in these fatties. In contrast, fatties calorically restricted from birth responded at elevated rates at all ages compared to lean controls. At 20 weeks restricted fatties, which were as obese on a percentage body composition basis as ad lib fatties, responded significantly more than the latter group at all fasting levels. Metabolic measurements revealed elevated fat cell size, LPL activity/cell and insulin levels in both fatty groups at 15 weeks, and elevated fat cell number in these groups at 33 weeks. The possibility is discussed that enhanced lipogenic factors present during the rapid growth period combine with the metabolic demands of growth to produce a “compensatory hyperphagia” in the fatty rat.     

The reduced proportion of New splenic T-cells in the zinc-deficient growing rat is not due to increased susceptibility to apoptosis

Dietary zinc deficiency has been associated with an increased risk of infection. It has been reported that zinc-deficient rats have fewer New T-cells (TCRαβ⁺CD90⁺) compared to diet-restricted and control rats, which over time could adversely affect the ability of the organism to fight off infections. We hypothesized that the lower proportion of New T-cells in zinc deficiency is due to an increased susceptibility to apoptosis. Weanling, Sprague Dawley rats were assigned to one of four dietary treatment groups for 3 weeks: zinc-deficient (ZD, <1 mg zinc/kg, ad libitum), diet-restricted (DR, 30 mg zinc/kg, limited to the amount of feed as consumed by ZD), marginally zinc-deficient (MZD, 10 mg zinc/kg, ad libitum) or control (CTL, 30 mg zinc/kg, ad libitum). Thymocytes and splenocytes were labeled for flow cytometric determination of cell surface markers and DNA staining (for simultaneous determination of the phenotype of apoptotic cells) and assessed by Western blotting for apoptotic markers. Cells were analyzed immediately, or after incubation for 7 h with or without dexamethasone. There was no difference in the proportion of CD90⁺ thymocytes; however ZD rats had a higher proportion of Cytotoxic (CD90⁺4⁻8⁺) thymocytes compared to MZD and CTL. ZD had a lower proportion of splenic New T-cells compared to DR, MZD and CTL. There was no effect of diet on the proportion of apoptotic thymocytes or splenocytes, except ZD splenoctyes had a lower Bax/Bcl-xl ratio compared to DR and CTL. We characterized the splenic New T-cells into Helper and Cytotoxic subsets and found that ZD had a higher ratio of Helper to Cytotoxic New T-cells compared to MZD and CTL. These results do not support the hypothesis of increased apoptotic removal of New T-cells in ZD in growing rats. The regulation of CD90 expression should be explored in future studies.     

Estradiol and progesterone suppress feeding induced by 2-deoxy-D-glucose

The feeding response to 2-deoxy-d-glucose (2DG) was measured in gonadally intact females (NORM) during estrus (EST) or diestrus (DIES) and in ovariectomized (OVEX) rats following injections of estradiol benzoate and progesterone (HORM) or oil (OIL). In both groups the response to 2DG under conditions of EST or HORM was significantly suppressed relative to DIES or OIL. The suppressant effect of hormones on 2DG feeding was not accounted for by the suppressant effects of ovarian hormones on ad lib feeding or a function of the body weight of the subject. The results are discussed in the context of ovarian hormonal effects on glucostatic control of feeding.     

Sham feeding is inhibited by dietary-induced obesity in rats

A group of six female, albino rats were maintained on a cafeteria diet of cookies, milk, and elevated-fat (shortening), rat-chow mixture and rat chow while a similar group received only rat chow ad lib for 17 weeks. When the groups differed significantly in mean body weight (obese-387.5 g, controls-287.2 g; p less than 0.001), gastric fistulas were implanted in each animal. After recovery, the rats were adapted to a liquid diet and assessed for sham feeding. Control-fed, normal-body-weight subjects showed substantial sham feeding when ingesting the Vivonex with the fistulas open compared to fistula-closed intake; meal frequency, meal size (apart from the initial meal) and total food intake were significantly increased while the satiety ratios following each meal were significantly decreased. Obese animals showed no significant increased feeding and satiety ratios were unreliably altered; while normal-body-weight controls increased 4-hr food intakes by 93% and halved their mean satiety ratios the obese animals showed an 8% increase in 4-hr food intake and only a 22% decrease in mean satiety ratios. We offer the hypothesis that, when animals are induced to become obese by palatable and varied diets which are then terminated, the anorexia produced is independent of gastrointestinal interactions inasmuch as that anorexia extends to sham feeding.     

Passive avoidance performance of mice fed marginally or severely zinc deficient diets during post-embryonic brain development

Swiss-Webster outbred mice were fed marginally or severely zinc deficient diets (9 or 2 ppm zinc) from day 16 gestation to day 15 postnatal. Control mice were fed a 100 ppm diet either ad lib or in amounts equal to the diet intake of deprived mice (pair fed controls). Male and female offspring were tested at 70 days of age in a one-trial passive avoidance task with a 30 min train-test interval. Both marginally and severely zinc deprived offspring (but not pair fed controls) had shorter avoidance latencies than offspring of ad lib fed zinc replete controls. Zinc deprived offspring did not differ from control mice when either baseline or “stressed” (exposure to novel environment) plasma corticosterone levels were quantitated. Further, zinc staining patterns of the hippocampus (Timm-sulfide stain) were not altered in the nutritionally deprived offspring. Thus marginal dietary zinc deficiency during development can lead to impaired passive avoidance performance in adult mice. This behavioral effect is not associated with altered pituitary-adrenocortical activity or with a permanent reduction in hippocampal zinc staining. This result has significant implications for the influence of zinc deprivation in utero and in the neonatal animal on adult behavior characteristics.     

Returning 'cafeteria-fed' rats to a chow diet: negative contrast and effects of obesity on feeding behaviour

Two experiments examined the feeding behaviour and body weight changes of rats returned to a standard chow diet after prolonged periods of cafeteria feeding. In the first, both overweight (ad lib fed) and 'normal' weight (fed a restricted ration of cafeteria foods) cafeteria rats became hypophagic compared to chow-fed rats following their return to chow feeding. However, the overweight rats' hypophagia was initially more severe and was longer lasting. In the second experiment, analysis of meal patterns recorded throughout the first 4 weeks on chow after 26-32 weeks of ad lib cafeteria feeding showed that the hypophagia was due to a reduction in mean meal size (MMS) and meal frequency (MF). Food intake and MMS subsequently recovered to within control levels (by 2-3 weeks), but MF remained persistently low. There was a decrease and then a recovery in eating rate (ER) which paralleled the changes in MMS. The previously cafeteria-fed rats lost only 60% of their excess body weight. These findings are interpreted in terms of a negative contrast effect (changes in MMS and ER) and an inhibitory action of increased adiposity on feeding (affecting mainly MF).     

Low body temperature in long-lived Ames dwarf mice at rest and during stress.

Among homeothermic animals, larger species generally have lower metabolic rates and live longer than do smaller species. Because Ames dwarf mice (dwarfs) live approximately 1 year longer than their larger normal sex- and age-matched siblings (normals), we hypothesized that they would have lower body core temperature (Tco). We, therefore, measured Tco of six dwarfs and six normals during 24-h periods of ad lib feeding, 24-h food deprivation, and emotional stress induced by cage switching. With ad lib feeding, Tco of dwarfs averaged 1.6 degrees C lower than normals; during food deprivation, Tco of both dwarfs and controls was significantly lower than when food was available ad lib; and following cage switch, Tco was elevated in both groups. However, during all three experiments, Tco was significantly lower in dwarfs than in normals. These data support the hypothesis that Ames dwarf mice, which live longer than normal size controls, maintain lower Tco than normals. Because dwarfs are deficient in thyroid stimulating hormone (TSH) and growth hormone (GH), their low Tco may be a result of reduced thermogenesis due to lack of those hormones. However, whether low Tco per se is related to the increased longevity of the dwarf mice remains an interesting possibility to be investigated.     

Circadian patterning of feeding,drinking and activity during diurnal food access in rats

Using an “ecologically” relevant approach, the present study investigated (1) the association between feeding and drinking patterns and their circadian organization and (2) changes in general activity (or life-style), during ad lib conditions (fa:LD) and in a situation where access to food was restricted to the light phase (fa:L). Rats were housed in large outcages with nest boxes. Feeding, drinking, activity, outcage and nest occupation were recorded automatically throughout the day-night cycle. Access to food was restricted by a sliding door situated in front of the food hopper. Under ad lib conditions rats were mainly nocturnal, eating 94% and drinking 95% of their daily intakes at night. The patterns of food and water intake were similar, showing a bimodal distribution over the dark phase. During fa:L rats showed an initial large peak in feeding with lights on, followed by a long pause. Thereafter, feeding activity was variable but remained at a low level. The nocturnal drinking pattern persisted. However, 17.5% of daily water intake was meal-associated, compared with 71% during ad lib. Diurnal activity was associated with feeding and nocturnal activity with drinking. Nocturnal outcage and nest occupation patterns were not shifted to the light phase. The experiment demonstrates that rats on an fa:L schedule reduce food and water intake, and body weight, while still retaining circadian characteristics in the temporal distribution of drinking, activity, outcage and nest occupation. Further, although feeding and drinking may be causally related they need not occur in close temporal association. The rapidity of re-establishment of the normal feeding pattern, on return to free-feeding, and the close association with drinking under normal conditions, suggest the existence of a single or two coupled oscillators controlling feeding and drinking.     

Zinc-deficient diet aggravates ventilation-induced lung injury in rats

We investigated the effects of zinc deficiency on acute lung injury(ALI)induced by mechanical ventilation.Male Sprague-Dawley rats were fed with a zinc-deficient or zinc-proficient diet for 4 weeks,and then received mechanical ventilation at normal frequency and pressure for 30 min.Total protein,cell count,the number of polymorphonuclear neutrophil(PMN)in the bronchoalveolar lavage(BAL),and vascular endothelial growth factor(VEGF)expression in the lung were determined.Activation of nuclear factor-κB(NF-κB)was detected by examining the phosphorylation of NF-κB(pNF-κB p65)and the expression of inhibitor of NF-κB(pI-kBα).Compared to the controls,total cell count and the number of PMNs were significantly increased to 160% and 140%,respectively,in zinc-deficient rats treated with ventilation.Activation of NF-κB was significantly increased and VEGF was also increased to three-folds.Zinc deficiency aggravated the inflammatory response in rats and was associated with the overexpression of VEGF in response to mechanical ventilation.Zinc supplementation may be beneficial to zinc-deficient patients during mechanical ventilation.     

Maternal deprivation in neonatal rats alters the expression of circadian system under light-dark cycles and restricted daily feeding in adulthood

Effects of maternal deprivation (MD) with different conditions were examined on the circadian rhythms in plasma corticosterone and locomotor activity in adult rats under ad libitum and restricted daily feeding (RF), in which rats had free access to food for 2 h for 3 weeks. Three different types of MD were performed from postnatal day 1 (P1) to day 6 (P6); MD for 12 h/day (MD12), for 3 h/day in the morning (MD3am) and in the afternoon (MD3pm). Under ad libitum feeding, corticosterone levels at 08:00 h and 24:00 h were significantly increased in MD12 rats. A similar increase was detected in MD3am rats, but not in MD3pm rats. Under the RF, corticosterone levels at 08:00 h, 24:00 h, and 04:00 h were significantly increased in MD12 rats. Similar elevations were detected in MD3am rats, but not in MD3pm rats. Furthermore, prefeeding locomotor activities under the RF increased significantly in MD12 rats. A similar increase was observed in MD3am rats, but not in MD3pm rats. Keeping ambient temperature and humidity at 37 degrees C and 70-80% during MD attenuated the effects of MD on the corticosterone rhythmicities under RF as well as ad lib feeding. It is concluded that MD in the early postnatal period influences the expressions of circadian rhythms in plasma corticosterone and locomotor activity in adult rats. Not only the length but also the time of day of MD seems to be critical for these effects. Ambient temperature and/or humidity are suggested to be one of the factors by which MD exerts its effect.     

锌对大鼠脾脏淋巴细胞功能的影响

通过大鼠锌（ＺＤ），过量锌（ＺＥ）模型，研究３锌对脾脏Ｔ、Ｂ淋巴细胞功能的影响，结果：①ＺＤ、ＺＥ组的脾重及脾脏指数，与对喂（ＰＦ）组、自由喂养（ＡＬ）组，缺锌后再补锌（ＺＤ＋ＡＬ）组比较，差别非常显著。②ＺＤ、ＺＥ组外周血白细胞减低，以淋巴细胞为主。③ＺＤ、ＺＥ组Ｔ、Ｂ淋巴细胞增殖率非常显著低于ＰＦ、ＡＬ和ＺＤ＋ＡＬ组。④ＺＤ、ＺＥ组的总脾淋巴细胞，Ｔ、Ｂ淋巴细胞细胞周期Ｇ０／Ｇ１％增高，而Ｓ％     

Circadian rhythms of blood pressure and heart rate in conscious rats: effects of light cycle shift and timed feeding

The aim of the present study was to evaluate the impact of a shift in the light cycle and of restricted food availability on circadian rhythms of blood pressure, heart rate, and behavioral activity in freely moving rats by radiotelemetry. In rats that were fed ad lib, a shift of the light cycle by 6 h (from lights on 0700-1900 h to lights on 1300 to 0100 h) induced an immediate, but from then on gradual, shift of the circadian rhythms of blood pressure, heart rate, and behavioral activity, which took 4-5 days to fully synchronize with the new light cycle. Rats on a normal light cycle receiving feeding for 1 h only during the light period rather than ad lib feeding, showed a suppressed circadian rhythm, with the dark period values reduced to values not significantly different from those observed in the light period. In addition, during the timed feeding blood pressure, heart rate, and behavioral activity peaked to levels that were normally seen during the dark period. These data show that environmental factors such as timed feeding or changes in the light cycle have a marked influence on the circadian rhythms of blood pressure and heart rate.     

Restoring the metabolic disturbance of zinc: may not only contribute to the prevention of esophageal squamous cell cancer

The relationship between zinc deficiency (ZD) and esophageal squamous cell cancer (ESCC) is well-established. Zinc supplement has long been considered as potential strategy for prevention of ESCC. However, the effect of zinc replenishment on prevention of ESCC is still in controversy and no convincing evidence to support these proposals has become known. We hypothesized that zinc deficiency might be the major initiator for dysfunction of numerous genes and subsequent carcinogenesis of esophagus, and replenishment of zinc might improve the prognosis of patients with ESCC.     

Comparison of the interoceptive sensory consequences of CCK, LiCl, and satiety in rats

In three experiments we assessed the degree to which ad lib feeding, injection of cholecystokinin (CCK), and injection of lithium chloride (LiCl) produce states with similar sensory consequences. In each experiment, two groups of rats were trained to use cues arising from food deprivation and satiation as discriminative signals for shock. One group was shocked when deprived but not when nondeprived. The other group received the reversed discrimination. Testing began when incidence of freezing was greater under the shocked deprivation than under the nonshocked deprivation condition. In Experiment 1, the rats were tested under 24-hr food deprivation after injections of CCK, LiCl, and saline (in counterbalanced order). We reasoned that if either CCK or LiCl induce satiety-like states, they should promote patterns of responding different from those produced by saline but similar to those produced by ad lib feeding. The effects of CCK on freezing did not differ from those of saline, whereas both CCK and LiCl had effects that were different from ad lib feeding. This pattern of results was also obtained when deprivation level during training and testing was reduced to 8 hr (Experiment 1A) and also when rats received small amounts of food in conjunction with CCK (Experiment 2). The intubation of a high-calorie stomach load (Experiment 1A) produced a response profile like that observed after free feeding. Freezing after LiCl treatment differed from that observed after free feeding and from that found after injection of CCK. The results indicate that rats can differentiate between the sensory consequences of the states produced by CCK, by LiCl, and by ad lib feeding.     

Plasma ammonia and liver ornithine transcarbamoylase activity in zinc-deficient rats

The effects of zinc deficiency on the activity of hepatic ornithine carbamoyltransferase (OCT) and plasma ammonia were studied in rats. One group received (ad libitum) zinc-deficient diet containing 2 ppm zinc and the other group received a diet containing 110 ppm zinc (group pair-fed control) equal to the amount consumed by zinc-deficient rats during the previous 24 h. Rats were killed at weekly intervals. Blood urea nitrogen (BUN), plasma ammonia, and hepatic OCT activity were determined. By end of the 1st wk on zinc-deficient diet, the plasma ammonia levels became significantly higher than those of the controls and remained elevated thoughout the study period. BUN increased initially for 2 wk in the deficient rats, but by the end of 4 wk the levels were lower than in the controls. The hepatic OCT activity in deficient animals was significantly lowered as compared to the controls by the 3rd wk. It is concluded that an increase in plasma ammonia may occur as a result of deficiency of zinc.     

Intra-uterine nutrition and its effects on aggression

Prenatal zinc deficiency and prenatal undernutrition were found to have adverse effects on the food consumption and weight gain of pregnant dams and their offspring. Pups whose dams suffered prenatal zinc deficiency (ZD) consumed less food and gained less weight than pups whose dams suffered prenatal undernutrition (PF). The PF pups consumed less food and gained less weight than pups whose dams were normally fed (AL). The ZD females at age 75 days were significantly more aggressive than the PF females, while the PF females were more aggressive than the AL females. At age 105 days, ZD females were significantly more aggressive than the PF and AL females. There were no differences in aggression between the PF and AL females at 105 days. Among the ZD, PF, and AL male offspring, there were no differences in aggression at either age level except that the 75 day old PF males were significantly less aggressive than the AL males. Thus prenatal malnutrition, especially zinc deficiency, seems to have differential effects on the aggressive tendencies of female and male offspring.     

Schedule and quinine induced deprivation in feeding and refeeding

Twenty female albino rats were adapted to either 0 or 23 hr of food deprivation. Half of each group was then fed 0.125% quinine sulfate adulterated diet for seven days. Following the quinine feeding, ad lib feeding (refeeding) was instituted for 14 days. Several conclusions were drawn from the results: (1) rats on a deprivation schedule fail to show a predicted change to regulation on the basis of taste rather than calories; (2) rats on food deprivation actually increase their relative intake of water; (3) refeeding after a deprivation schedule does not lead to depression of initial intake below normal, but otherwise the process of recovery follows the same course as after total starvation.