Regulation of food intake in monkeys: Response to caloric dilution

Ability to regulate level of energy intake was studied in adult rhesus monkeys (Macaca mulatta) using calorically diluted diets. Twenty-four hour access to a complete liquid diet was provided via leakproof gravity feeders. The addition of water provided 4 caloric concentrations over the range of 0.5 to 1.35 kcal/ml. Average caloric intake per kg body weight was 84 ± 0.7 kcal/kg (mean ± SE). Seven of the eight monkeys maintained a constant caloric intake by adjusting oral intake in response to randomly ordered but sustained changes in caloric density. One monkey ingested a significantly higher caloric load while receiving the highest density diet. Rates of compensation for dilution following each diet change varied widely, occurring over periods of 3 days to 2 weeks. It was concluded that individual monkeys vary significantly in the rat of adjustment to caloric dilution, and thus long term studies must be used in studying controls of feeding in monkeys.     

Age-related decline in striatal volume in rhesus monkeys: assessment of long-term calorie restriction

Using magnetic resonance imaging (MRI), we measured striatal volume in 22 male rhesus monkeys undergoing calorie restriction (CR) for 11-13 years and 38 monkeys who were fed ad libitum (CON). CR delays the onset of many age-related processes, and this study tested whether it would alter the age-related decline in striatal volume. The CON and CR groups were sub-divided into middle age (less than 24 years old) and old age groups. Contrary to expectation, volumes of the putamen (not the caudate nucleus) were larger bilaterally in the CON than in the CR group both at middle age and senescence. Regression analysis (region volume versus age) indicated bilateral age-related declines in putamen and caudate nucleus volumes in the old CON monkeys, but only for the putamen in the old CR monkeys. Because tests for slopes found no differences between the groups, the data do not establish an effect of CR. Further study, involving sequential imaging, is warranted in order to clarify the possible effects of CR on age-related changes in striatal volume.     

Quantifying food intake in socially housed monkeys: social status effects on caloric consumption

Obesity results from a number of factors including socio-environmental influences and rodent models show that several different stressors increase the preference for calorically dense foods leading to an obese phenotype. We present here a non-human primate model using socially housed adult female macaques living in long-term stable groups given access to diets of different caloric density. Consumption of a low fat (LFD; 15% of calories from fat) and a high fat diet (HFD; 45% of calories from fat) was quantified by means of a custom-built, automated feeder that dispensed a pellet of food when activated by a radiofrequency chip implanted subcutaneously in the animal's wrist. Socially subordinate females showed indices of chronic psychological stress having reduced glucocorticoid negative feedback and higher frequencies of anxiety-like behavior. Twenty-four hour intakes of both the LFD and HFD were significantly greater in subordinates than dominates, an effect that persisted whether standard monkey chow (13% of calories from fat) was present or absent. Furthermore, although dominants restricted their food intake to daylight, subordinates continued to feed at night. Total caloric intake was significantly correlated with body weight change. Collectively, these results show that food intake can be reliably quantified in non-human primates living in complex social environments and suggest that socially subordinate females consume more calories, suggesting this ethologically relevant model may help understand how psychosocial stress changes food preferences and consumption leading to obesity.     

Controlling caloric consumption: protocols for rodents and rhesus monkeys.

One approach for investigating biological aging is to compare control-fed animals with others restricted in calorie intake by 20% or more. Caloric restriction (CR) is the only intervention shown to extend the maximum lifespan of several invertebrates and vertebrates including spiders, fish, rats and mice. The capacity of CR to retard aging in nonhuman primates is now being explored. The rodent studies show that CR opposes the development of many age-associated pathophysiological changes, including changes to the brain and changes in learning and behavior. One goal of studying CR in rodent is to determine the mechanisms by which it retards aging to design interventions that duplicate those effects. The methods that we use for conducting CR studies on mice and rhesus monkeys are described. We employ procedures designed to achieve a high degree of caloric control for all animals in the study. As used in our studies, this control includes the following features: 1) animals are individually housed, and 2) all individuals in the control group eat the same number of calories (i.e., they are not fed ad lib). Although this method results in strict caloric control for all animals, there seems to be considerable procedural flexibility for the successful conduct of CR studies.     

Food intake and the menstrual cycle in rhesus monkeys

Fourteen adult female rhesus monkeys were observed for 1 complete menstrual cycle, five of them for 2 cycles. Changes in LH, FSH, estrogens and progesterone were monitored daily. Mean hormonal concentrations followed patterns previously demonstrated in primates, with a typical late follicular phase estrogen peak preceeding the ovulatory LH and FSH surges. Maximal sexual skin color intensity paralleled the midcycle increase in estrogens. The results indicate that food intake fluctuated with changes in hormonal secretion. A significant decrease in the amount of food consumed correlated well with the midcycle estrogen and gonadotropin surges. The amount of food consumed during the luteal phase was greater than that of the early follicular phase.     

Control of food intake and meal patterns in monkeys

Feeding patterns have been studied in 10 adult male rhesus monkeys (Macaca mulatta) adapted to primate restraint chairs for physiological studies. Using an automated computer-monitored liquid diet feeding system, feeding behavior was studied under 8 hour and 24 hour feeding schedules. Calorie/kg intake was significantly reduced with the 8 hour schedule (p<0.05). Average rate of weight gain and feeding efficiency were also lower with 8 hour feeding, but not significantly different from 24 hour ad lib feeding (p>0.10). Meal size was negatively correlated with meal frequency across monkeys, but not within monkeys. Although monkeys ingested 78% of their voluntary intake during the light hours, there was no difference in meal size, meal duration, or rate of feeding between light and dark periods. With feeding restricted to 8 hours during the light period, meal size was positively correlated with the length of the interval preceding the meal in 4 of 6 monkeys, and meal size was also positively correlated with length of the postmeal interval in 3 of 6 monkeys. In contrast, 24 hour ad lib fed monkeys showed no relationship between premeal interval and subsequent meal size, but a positive correlation between meal size and postmeal interval in 3 of 4 monkeys. We conclude that feeding schedule or deprivation state may alter the relative roles of “hunger” and “satiety” signals in regulating food intake amounts and patterns.     

Homocysteine,neural atrophy,and the effect of caloric restriction in rhesus monkeys

Higher serum homocysteine (Hcy) levels in humans are associated with vascular pathology and greater risk for dementia, as well as lower global and regional volumes in frontal lobe and hippocampus. Calorie restriction (CR) in rhesus monkeys (Macaca mulatta) may confer neural protection against age- or Hcy-related vascular pathology. Hcy was collected proximal to a magnetic resonance imaging (MRI) acquisition in aged rhesus monkeys and regressed against volumetric and diffusion tensor imaging indexes using voxel-wise analyses. Higher Hcy was associated with lower white matter volume in pons and corpus callosum. Hcy was correlated with lower gray matter volume and density in prefrontal cortices and striatum. CR did not influence Hcy levels. However, control monkeys exhibited a strong negative correlation between Hcy and global gray matter, whereas no relationship was evident for the CR monkeys. Similar group differences were also seen across modalities in the splenium of the corpus callosum, prefrontal cortices, hippocampus, and somatosensory areas. The data suggest that CR may ameliorate the influence of Hcy on several important age-related parameters of parenchymal health.     

Cytokine responses in young and old rhesus monkeys: effect of caloric restriction.

Caloric restriction (CR) is the only known intervention demonstrated to retard a great variety of aging processes, extend median and maximum life-span, and decrease the incidence of age-associated diseases in mammals. Paralleling findings from rodent studies, studies in rhesus monkeys (Macaca mulatta) suggest that CR may retard many age-sensitive parameters in primates. A recent study in rhesus monkeys showed age-related dysregulation of cytokine levels. Specifically, age-related increases in interleukin-10 (IL-10) and IL-6 proteins were observed in supernatants from lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMCs), and interferon-gamma (IFN-gamma) protein exhibited an age-related decrease in phytohemagglutinin (PHA)-stimulated PBMCs. To investigate effects of CR on age-related changes in cytokine production, we obtained PBMCs from control and CR rhesus monkeys aged 6-7 and 22-25 years. We evaluated IL-10 and IL-6 protein and gene expression after exposure to LPS and IFN-gamma protein and gene expression after PHA stimulation. The results revealed significantly higher levels of IFN-gamma protein and gene expression in aged monkeys on CR for 2 years compared with controls. No significant CR effects were observed on IL-10 and IL-6 protein levels. IFN-gamma plays an important role in the initial defense mechanism against viral and microbial disease and cancer. Altered regulation of IFN-gamma in old CR rhesus monkeys may be a key factor in reducing cancer incidence and other age-associated diseases.     

Age-related changes in glial cells of dopamine midbrain subregions in rhesus monkeys

Aging remains the strongest risk factor for developing Parkinson's disease (PD), and there is selective vulnerability in midbrain dopamine (DA) neuron degeneration in PD. By tracking normal aging-related changes with an emphasis on regional specificity, factors involved in selective vulnerability and resistance to degeneration can be studied. Towards this end, we sought to determine whether age-related changes in microglia and astrocytes in rhesus monkeys are region-specific, suggestive of involvement in regional differences in vulnerability to degeneration that may be relevant to PD pathogenesis. Gliosis in midbrain DA subregions was measured by estimating glia number using unbiased stereology, assessing fluorescence intensity for proteins upregulated during activation, and rating morphology. With normal aging, microglia exhibited increased staining intensity and a shift to more activated morphologies preferentially in the vulnerable substantia nigra-ventral tier (vtSN). Astrocytes did not exhibit age-related changes consistent with an involvement in regional vulnerability in any measure. Our results suggest advancing age is associated with chronic mild inflammation in the vtSN, which may render these DA neurons more vulnerable to degeneration.     

Age-related decline in striatal volume in monkeys as measured by magnetic resonance imaging

Age-related declines in striatal markers for the dopamine system have been demonstrated in several species. The current study investigated structural changes during aging in the rhesus monkey striatum. Male monkeys were studied using a volumetric spoiled gradient recall (SPGR) magnetic resonance imaging protocol. The caudate nucleus and putamen were segmented by manual tracing using landmarks made in the orthogonal planes. The whole brain volume (defined as volume of gray and white matter plus cerebrospinal fluid in ventricles and sulci) was measured using a semi-automated algorithm. There was no correlation between age and whole brain volume. There were age-related declines in normalized (i.e. brain region/whole brain volume) caudate nucleus and putamen volumes. Monkeys in the young group (n = 7, 39-45 months old) had larger volumes of both the caudate nucleus and putamen than animals in the middle-age (n = 5, 120-180 months) or old (n = 7, 291-360 months) groups. The current results provide normative data to assess potential interventions (e.g. caloric restriction) in the aging process.     

Palatability and caloric density as determinants of food intake in hyperphagic and normal rats

Female albino rats with lesions of the ventromedial hypothalamus and without any lesions were presented with powdered diets, the caloric density and palatability of which were varied independently by adding kaolin and sodium saccharin or confectioner's sugar. As the dilution of the unsweetened diet increased above 20% the lesioned rats decreased food consumption more than did the controls, and they increased consumption less than did the controls over successive days of exposure to the diluted diets. Sweetening the dilute diets usually produced a larger increase in food consumption among the hyperphagic rats, especially those in the dynamic phase, than among their controls. The data are interpreted in terms of the hypothesis that lesions of the ventromedial hypothalamus remove or attenuate the long term influence of nutritional balance, leaving body weight to be adjusted to a set point defined by the interaction of prevailing values of other stimuli such as diet palatability that are known to influence initiation and termination of eating.     

Food intake and meal patterns in rhesus monkeys: significance of chronic hyperinsulinemia

To investigate the role of plasma insulin on food intake, we have examined the effect of naturally occurring chronic hyperinsulinemia on the feeding behavior of male rhesus monkeys. Two groups of monkeys, a group with normal fasting insulin concentrations (52.4 +/- 2.2 microU/ml) (mean +/- SE) and a hyperinsulinemic group (148.6 +/- 14.5 microU/ml), were selected to be similar in weight, 13.0 +/- 1.0 and 15.3 +/- 0.5 kg, respectively, prior to study. Food intake and feeding patterns were recorded and analyzed. No differences in either daily caloric intake, 815.2 +/- 27.4 versus 890.0 +/- 64.2 kcal (p less than 0.32), or feeding patterns were found. The number of meals taken per day did not differ between the two groups, 8.7 +/- 1.7 versus 6.7 +/- 1.1 (p less than 0.35), nor did meal size differ, 129 +/- 16.5 versus 110.5 +/- 16.3 (p less than 0.45). We conclude that chronic endogenous hyperinsulinemia as it occurs naturally in some obese rhesus monkeys has no significant effect on daily feeding behavior.     

Iron accumulation in the striatum predicts aging-related decline in motor function in rhesus monkeys

Changes in the nigrostriatal system may be involved with the motor abnormalities seen in aging. These perturbations include alterations in dopamine (DA) release, regulation and transport in the striatum and substantia nigra, striatal atrophy and elevated iron levels in the basal ganglia. However, the relative contribution of these changes to the motor deficits seen in aging is unclear. Thus, using the rhesus monkey as a model, the present study was designed to examine several of these key alterations in the basal ganglia in order to help elucidate the mechanisms contributing to age-related motor decline. First, 32 female rhesus monkeys ranging from 4 to 32 years old were evaluated for their motor capabilities using an automated hand-retrieval task. Second, non-invasive MRI methods were used to estimate brain composition and to indirectly measure relative iron content in the striatum and substantia nigra. Third, in vivo microdialysis was used to evaluate basal and stimulus-evoked levels of DA and its metabolites in the striatum and substantia nigra of the same monkeys. Our results demonstrated significant decreases in motor performance, decreases in striatal DA release, and increases in striatal iron levels in rhesus monkeys as they age from young adulthood. A comprehensive statistical analysis relating age, motor performance, DA release, and iron content indicated that the best predictor of decreases in motor ability, above and beyond levels of performance that could be explained by age alone, was iron accumulation in the striatum. This suggests that striatal iron levels may be a biomarker of motor dysfunction in aging; and as such, can be monitored non-invasively by longitudinal brain MRI scans. The results also suggest that treatments aimed at reducing accumulation of excess iron in the striatum during normal aging may have beneficial effects on age-related deterioration of motor performance.     

The effects of caloric dilution on meal patterns and food intake of ponies

In order to determine if horses will increase their intake in response to caloric dilution, four pony geldings were fed ad lib a mixed grain diet either undiluted (3.4 Mcal/kg of digestible energy) or diluted (wt/wt) with 25% sawdust (2.6 Mcal/kg) or with 50% sawdust (1.7 Mcal/kg). The mean daily caloric intake was 17,457 kcal (3.4 Mcal diet), 17,546 kcal (2.6 Mcal diet) and 12,844 kcal (1.7 Mcal). The mean time spent eating was 246 (3.4 Mcal), 351 (2.6 Mcal), and 408 (1.7 Mcal) minutes/day. Meal size increased and meal frequency decreased with increasing dilution. The median long survivorships of intermeal intervals were 6.4 min (3.4 Mcal), 3.95 min (2.6 Mcal) and 4.91 min (1.7 Mcal). Ponies responded to caloric dilution by increasing the volume of intake to maintain caloric intake when the diet had 25% diluent. When the diet was diluted by 50%, intake was increased, but not at a rate adequate to maintain caloric intake. However, the ponies were able to maintain body weight.     

Age-related cognitive deficits in rhesus monkeys mirror human deficits on an automated test battery

Aged non-human primates are a valuable model for gaining insight into mechanisms underlying neural decline with aging and during the course of neurodegenerative disorders. Behavioral studies are a valuable component of aged primate models, but are difficult to perform, time consuming, and often of uncertain relevance to human cognitive measures. We now report findings from an automated cognitive test battery in aged primates using equipment that is identical, and tasks that are similar, to those employed in human aging and Alzheimer's disease (AD) studies. Young (7.1+/-0.8 years) and aged (23.0+/-0.5 years) rhesus monkeys underwent testing on a modified version of the Cambridge Automated Neuropsychological Test Battery (CANTAB), examining cognitive performance on separate tasks that sample features of visuospatial learning, spatial working memory, discrimination learning, and skilled motor performance. We find selective cognitive impairments among aged subjects in visuospatial learning and spatial working memory, but not in delayed recall of previously learned discriminations. Aged monkeys also exhibit slower speed in skilled motor function. Thus, aged monkeys behaviorally characterized on a battery of automated tests reveal patterns of age-related cognitive impairment that mirror in quality and severity those of aged humans, and differ fundamentally from more severe patterns of deficits observed in AD.     

Age-related decline in immunity: implications for vaccine responsiveness

Aging is associated with declines in immune system function, or 'immunosenescence', leading to progressive deterioration in both innate and adaptive immunity. These changes contribute to the decreased response to vaccines seen in many older adults, and morbidity and mortality from infection. Infections (e.g., influenza, pneumonia and septicemia) appear among the top ten most-common causes of death in adults in the USA aged 55 years and older. As immunosenescence has gathered more attention in the scientific and healthcare communities, investigators have demonstrated more links between immunosenescent changes and morbidity and mortality related to infections and declining vaccine responses. This review summarizes the recent literature on age-dependent defects in adaptive and innate immunity, data linking these defects to poor vaccine response and morbidity and mortality, current recommendations for vaccinations and potential strategies to improve vaccine efficacy in older adults.