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.     

Reductions in maternal food and water intake account for prenatal stress effects on neurobehavioral development in B6D2F2 mice

This study evaluated the role of stress-induced reductions in food intake in pregnant B6D2F1 mice in the production of developmental abnormalities in the offspring. One group of dams underwent one hour of physical restraint stress twice daily from days 12 to 17 of gestation. A second group was not restrained but, during this period, each dam was pair-fed to a weight-matched partner in the stressed group. A third group was left undisturbed and allowed unlimited access to lab chow and water. The restraint stress procedure reduced the average daily food and water intake in the dams, resulting in lower maternal weights, with the unstressed pair-fed group being affected more severely than the stressed group. The offspring of both the stressed and pair-fed dams were lighter than the offspring of the dams fed ad lib on day 20 postconception, and exhibited lower brain weights on day 32 postconception. Neurobehavioral development, as assessed on a battery of sensorimotor tests on day 32 postconception, was retarded in the prenatally stressed pups in comparison to the ad lib pups, while the pair-fed pups did not differ from either group. There were no differences among any of the groups on day 50 body or brain weight, body or tail length, anogenital distance, or locomotion, rearing and defecation in the open field.(ABSTRACT TRUNCATED AT 250 WORDS)     

Hypothalamic hyperphagia despite imposed diurnal or nocturnal feeding and drinking rhythms.

Rats normally do most of their eating at night. When ad lib fed rats are made hyperphagic with lesions or parasagittal hypothalamic knife cuts the increases in eating occur primarily during the day. This suggests that a disruption of circadian rhythms may mediate the overeating. However, when knife cut rats were food and water deprived all day excessive eating occurred at night. Similarly, when they were deprived all night overeating occurred during the day. Under both conditions od deprivation the food intakes and rapid weight gains of the ad lib fed knife cut group were defended. It was concluded that: (1) in hypothalamic hyperphagia either the excessive food intake or the excessive weight gain is defended when food and water are available only half of each day, and (2) disruption of nocturnal feeding and drinking rhythms is not the cause of hypothalamic hyperhagia.     

Maternal malnutrition in the rat: Effects on food intake and body weight

The effects of dietary protein level on food intake and body weight were examined in adult female rats during a 35-day pre-mating period and during gestation and lactation. During the pre-mating period, no differences in daily food intake were observed among rats fed a 6% casein, 8% casein or 25% casein diet. However, during this period, rats fed the 6% casein diet gained significantly less weight than those with ad lib access to the 8% or 25% casein diets or than rats pair-fed the 25% casein diet in amounts equivalent to that consumed by rats in the 6% or 8% casein groups. Additionally, rats fed the 6% casein diet displayed decreased efficiency of energy utilization, calculated as weight gain per 100 kilocalories consumed, relative to rats fed the 8% or 25% casein diets. No differences in food intake were observed among the groups during gestation. However, rats fed the 6% casein diet gained less weight than rats fed the 8% or 25% casein diets. During lactation rats fed either the 6% or 8% casein diet consumed significantly less food than animals given the 25% casein diet ad lib. During the second week of lactation, rats receiving ad lib access to the 25% casein diet gained weight while those receiving the 6% or 8% casein diets continued to lose weight. At parturition, body weights of pups did not differ as a function of dietary condition. However, by day 12 of life, pups whose dams had ad lib access to the 25% casein diet weighed significantly more than pups whose dams consumed the 6% or 8% casein diet or whose dams were pair-fed the 25% casein diet in amounts equivalent to those consumed by rats fed the 6% or 8% casein diet.     

Synergistic effects of T-2 toxin and a low protein diet on erythropoiesis in mice

Young male Swiss mice fed on a semipurified diet containing 8% protein and 10 ppm of T-2 toxin developed erythroid hypoplasia within 2 weeks. Red blood cell counts declined to 36% of control values by 6 weeks but had risen to 45% of control values by 8 weeks. Between 4 and 8 weeks, erythropoietic tissues regenerated and reticulocyte counts became greatly elevated. The toxin-free semipurified diet was adequate for normal growth and did not cause anemia in control mice fed either ad lib, or at a restricted rate. Anemia did not occur in mice fed the 10-ppm level to T-2 toxin in either a semipurified diet containing 16% protein, or in a balanced natural-ingredient mouse diet. These observations demonstrated that the inhibitory effect of T-2 toxin in erythropoiesis in mice was transient, and depended on the nutritional composition of the diet.     

Effects of prenatal nutrition on learning and motivation in rats.

The results of two learning experiments revealed that adult male rats whose dams suffered either prenatal zinc deficiency or undernutrition showed, as compared to rats whose dams were fed ad lib, normal diets during pregnancy (1) more resistance to extinction following partial reinforcement, and (2) a faster occurring negative contrast effect when shifted from large to small reward. An incentive-motivation interpretation of the data suggests that the nutritionally deprived and normal animals differ with respect to the incentive value of the consummatory reward. This effect appears to be long-term.     

Effects of paternal and maternal undernutrition on growth of offspring in rats

In adult males, undernutrition is associated with altered spermatogenesis. The present study was performed to evaluate the possible role of paternal undernutrition on growth and development of rat offspring and to compare those effects with undernutrition in pregnant animals. Male rats were fed 100%, 80% or 60% of their ad lib food intake for eight weeks and were then bred to untreated females. Pregnant rats were mated with untreated males and received their food allotments throughout pregnancy. Although male body weights and rate of successful matings were decreased by paternal undernutrition, offspring litter size, birth weights, and weights at 21 days were not significantly altered. By contrast, maternal undernutrition was associated with decreased maternal weight gain, lower offspring birth weights and body weights at weaning.     

Duration of paternal alcohol consumption does not influence offspring growth and development

Three groups of male mice consumed liquid alcohol diets containing 25%, 10% or 0% ethanol derived calories (EDC) ad lib. Two additional groups were pair-fed to two of these groups, one to the 25% EDC group and one to the 10% EDC group. This pair feeding procedure was performed on an individual basis creating a dyad in which each individual in the pair received the same amount of calories but the source of the calories for alcohol varied. Males were bred to non-treated females after 1-2, 3-4 or 6-7 weeks of consumption. Male fertility was not affected by alcohol consumption. Offspring sired by alcohol consuming males did not differ from either ad lib or pair-fed controls in litter size, birth weight or weight at weaning regardless of duration of paternal alcohol consumption. These results indicate that neither duration of paternal alcohol consumption nor alcohol-induced undernutrition affects these gross morphological parameters in offspring.     

Learning deficits in first generation OF1 mice deficient in (n-3) polyunsaturated fatty acids do not result from visual alteration.

The effects of (n-3) polyunsaturated fatty acids (PUFA) diet deficiency on learning, electroretinogram and retinal fatty acid composition were assessed for the first time in OF1 mice. Pups fed the same diets (deficient in alpha-linolenic acid or a control) as their dams were used aged 7 weeks for passive avoidance test and fatty acid analysis of retinal phospholipids. Visual function was measured by electroretinography in 4- and 7-week-old mice. The (n-3) PUFA-deficient diet significantly decreased learning performance and retinal docosahexaenoic acid level in adult mice. The electroretinogram showed a significant alteration of b-wave amplitude in deficient mice at 4 weeks but not at 7 weeks. These results show that learning deficits in mice fed a diet deficient in (n-3) PUFA were not due to visual alteration.     

Arousal defect in L-phenylalanine fed rats

Rats fed a 5% L-phenylalanine diet from 28 days of age were tested 33 to 44 days later (61–72 days old) for spontaneous alternation, habituation to auditory stimulation, and passive avoidance. In spontaneous alternation and passive avoidance tests there was no significant difference from control rats. Habituation was faster than in the controls, interpreted as an inability to sustain arousal and attention. This may be a basic behavioral deficit in L-phenylalanine fed rats, accounting for deficits reported in more complex learning tests. Phenistix tests for phenylpyruvic acid in the urine were positive only during the first 4 weeks the 5% diet was fed. The behavioral deficit was present even after this period. To reestablish positive Phenistix tests the excess L-phenylalanine in the diet had to be raised to 10%. This diet severely affected the nutrition of the animals.     

Oral low-carbohydrate alcohol liquid diet induces experimental steatohepatitis in the rat

The intragastric tube feeding model of alcoholic liver disease in the rat induces significant liver histopathology, including steatohepatitis and fibrosis. The question is, if the same low-carbohydrate diet is fed ad lib, will the same pathology develop? Rats were fed a liquid diet with ethanol ad lib that was low in calories derived from carbohydrates for 2 months. The urinary ethanol levels (UALs) were monitored at hourly, daily, and weekly intervals, and the growth of the rats was charted. The liver histopathology and blood transaminase levels were determined. Rats fed ethanol grew 1 g/day, which was 2 g/day less than when they were fed the same diet intragastrically. UALs varied hourly between 150 and 500 mg%, daily between 120 and 360 mg%, and weekly between 0 and 500 mg%. Individual rat UALs showed no predictable pattern. The pair-fed controls ate all of their daily ration within 12 h, then fasted until the next day. The histopathology and blood alanine aminotransferase were similar to those seen with the intragastric tube feeding of the same diet, except that necrosis, inflammation, and fibrosis did not develop. The conclusion was that the oral feeding of a low-carbohydrate diet produces less liver injury than that produced by the same diet fed intragastrically. The UALs varied hourly, daily, and weekly in individual rats, making it difficult to synchronize UALs at the time of sacrifice.     

Zinc requirement for macrophage function: effect of zinc deficiency on uptake and killing of a protozoan parasite.

The effects of suboptimal levels of zinc, an essential trace element, on the ability of murine macrophages to associate with and destroy a pathogenic parasite, Trypanosoma cruzi, were evaluated. Young adult A/J mice were fed zinc-deficient, zinc-adequate or restricted amounts of a zinc-adequate diet for 28 days. On the basis of weight loss and parakeratosis, the zinc-deficient mice were further divided into moderately and severely zinc-deficient on Day 28. Both the percentage of mouse peritoneal macrophages (MPM) with associated parasites and the number of parasites per 100 macrophages were significantly lower for macrophages from moderately and severely deficient mice compared to MPM from mice fed restricted or zinc-adequate diet. Furthermore, MPM from both zinc-deficient groups of mice killed fewer internalized parasites than did MPM from restricted or zinc-adequate mice. Pretreatment of MPM from zinc-deficient mice with 5 micrograms zinc/ml for 30 min completely restored both their capacity to take up and kill the parasites. Other trace metals tested, including copper, manganese and nickel, failed to reverse the effects of zinc deficiency. These results point to an important role for zinc in the biochemical events associated with macrophage uptake and killing of the parasite.     

Developmental delays in offspring of rats undernourished or zinc deprived during lactation

Offspring of rats who were zinc or calorie deprived during lactation were administered a battery of reflex and motor tests from postnatal Day 4 to Day 21. Compared to offspring of ad lib-fed control rats, both zinc deprived and undernourished offspring exhibited developmental delays in reflexes which appeared after the first postnatal week (auditory startle, air righting, and rope descent). As the deficiencies continued the delays appeared to be more pronounced. The zinc deficiency did not add to the deficits associated with calorie restriction alone because there were no significant differences between the zinc deficient and undernourished pups on any of the measures except eye opening. When rehabilitated offspring were tested at 45 and 60 days of age for motor deficits there were no significant impairments resulting from preweaning dietary conditions. However, the growth retardation of zinc deprived and undernourished rats persisted long after dietary rehabilitation was implemented.     

Differential response of synaptic zinc levels to sensory deprivation in the barrel cortex of young and adult mice

The distribution of synaptic zinc after short-term (up to 48 h) tactile deprivation of vibrissae was investigated in the barrel cortex of mice using histochemical staining. In adult mice, 12 h after trimming selected rows of vibrissae, an increase in zinc staining in the deprived barrels was observed. This increase was still present 48 h after trimming. These results indicate that the level of synaptic zinc is rapidly regulated by neuronal activity in adult mice. In young (8-day-old) mice, the short-term deprivation did not alter zinc staining and only chronic sensory deprivation produced an increase in zinc staining. However, after long-term deprivation no changes were found in adult mice. These results suggest that different mechanisms might be involved in functional reorganization of zinc containing terminals in young and fully mature cerebral cortex.     

Effects of age and dietary restriction on the kidney glomeruli of mice: Observations by scanning electron microscopy

A study was undertaken to examine, by scanning electron microscopy (SEM), the kidney glomeruli of control mice 1 mo, 10 mo, and 24 mo of age, as well as dietarily restricted mice 10 mo and 24 mo of age. One month old female C57BL/6J mice were offered one of the following: (1) a control diet containing 24% protein fed ad lib; (2) the control diet fed on alternate days (intermittently fed); or (3) a diet containing 4% protein fed ad lib. Animals were sacrificed, by aldehyde perfusion at 1 mo, 10 mo, and 24 mo of age. The kidneys were sliced and prepared for SEM. There was a significant age-related increase in glomerular diameter and amount of microvilli on the podocyte surface (microvillus index). Although the diameters of the podocytes increased approximately 20% with age, these differences were not statistically significant. Feeding a 4% protein diet resulted in smaller diameters of glomeruli and podocytes as well as smaller microvilli indices as compared to those of control animals. Although similar differences were observed in the kidneys of intermittently fed animals, only the microvillus index was statistically significant. Therefore, dietary manipulations, which have been shown to increase life span, result in marked morphological differences when compared to control animals.     

Prenatal ethanol and stress in mice: 1. Pup behavioral development and maternal physiology

On days 12 to 17 of pregnancy, B6D2F1 mice were pair-fed liquid diets containing either 25% ethanol-derived calories or an isocaloric amount of maltose-dextrin. During this period, half the mice in each dietary condition also underwent two daily one-hour periods of restraint stress. A fifth group, given lab chow and water ad lib, was left undisturbed throughout gestation. Neither treatment affected offspring body weight on days 22 or 32 postconception, but undernutrition produced by the pair feeding procedure reduced day 32 body weight in all groups relative to the ad lib-fed group. Both prenatal ethanol and pair feeding led to delayed neurobehavioral development on day 32, while prenatal stress significantly reduced the degree of developmental delay caused by these factors. In a second study, restraint stress significantly reduced blood alcohol concentrations in pregnant dams on day 15 of gestation while elevating plasma corticosterone concentrations, and this elevation was consistent regardless of the dietary condition of the dam. The pair feeding procedure also produced corticosterone elevations but the effect of ethanol was not significant. These results suggest that prenatal stress in the presence of other physiological insults may act to counter the actions of those insults.     

The effects of intermittent dietary restriction on weight gain and body fat in white-footed mice, Peromyscus leucopus

The effects of repeated food restriction-refeeding cycles were investigated in second generation captive Peromyscus leucopus. Mice were maintained on 50% of their individual predetermined ad lib diet for 5 days followed by 10 days of refeeding ad lib. Cycles were repeated 3 times. When compared with controls maintained ad lib for the same time period, no significant differences in final weight or fat content were observed. Restricted-refed mice were, however, observed to both lose and regain weight at increasing rates with each subsequent cycle.     

Oscillatory changes in muscle lipoprotein lipase activity of fed and starved rats.

Lipoprotein lipase activity was measured at short time intervals in cardiac and skeletal muscles of normal and streptozotocin-treated diabetic rats fed ad libitum or deprived of food. In normal animals fed ad libitum, lipoprotein lipase activities of heart, diaphragm, soleus, and fast-twitch red fibers of the quadriceps muscle showed rhythmic oscillations that appeared to coincide with the nocturnal feeding habits of the animals. During the day (7 A.M. to 7 P.M.), when food consumption by the rats was greatly reduced, lipoprotein lipase activity in all muscles increased, followed by a decline to basal levels during the night. Similar oscillatory changes in lipoprotein lipase activity were observed in the muscles of diabetic rats fed ad libitum. In normal rats deprived of food, however, the oscillatory changes in muscle lipoprotein lipase activity were not abolished and persisted for at least 48 h. In diabetic rats starved during a 48-h period, the oscillatory changes in muscle lipoprotein lipase activity were markedly altered. In all animals, muscle lipoprotein lipase activities were not correlated to plasma glucagon levels.     

Effects of long-term restricted feeding on radial maze performance by aged rats

Rats were fed every other day (EOD) from 3-21 months of age prior to training in an 8-arm radial maze with retention intervals of 0, 1 or 5 hr imposed between the fourth and fifth choices. Their performance was compared to that of another aged group fed ad lib throughout life prior to radial maze training and to 3 month old controls. Rats fed ad lib until training adapted to the maze more slowly than young controls and were impaired in acquiring accurate spatial memory. EOD feeding eliminated age-related differences in the initial rate of adaptation to the radial maze but had no effect on the development of accurate spatial memory at any of the retention intervals. These results indicate that differences in deprivation history are probably not responsible for previous findings that radial maze training early in life prevents the appearance of age-related deficits in accurate spatial memory.