Body and organ mass in agouti and non-agouti deer mice (Peromyscus maniculatus).

Body, adrenal, brain, heart, liver, kidney, spleen and testis masses were determined for agouti and non-agouti deer mice (Peromyscus maniculatus gracilis) of both sexes. Body mass was highest for non-agouti females and lowest for agouti females; and sex differences in body mass were significant for agouti, but not non-agouti, deer mice. Adrenal, brain and liver masses were similar between color morphs; heart mass was greater in agouti males; and kidney, spleen and testis masses were all significantly greater for non-agouti deer mice. Splenomegaly in non-agouti deer mice was prominent, as spleens of non-agouti deer mice were 50% larger than those of agouti animals. Sex differences varied across organs and color morphs. For both color morphs, males had heavier adrenals and brains, whereas females had heavier livers and spleens. Kidney and heart mass was greater for female non-agouti deer mice, but for agouti animals, heart mass was greater in males and kidney mass differed little between the sexes. For both color morphs, testes and spleen mass was altered by photoperiod in 72 deer mice housed under short- or long-day conditions and the effect was stronger in non-agouti animals. This is the first report of splenomegaly and sex-specific body mass differences associated with the non-agouti allele.     

Thyroxine levels in agouti and non-agouti deer mice (Peromyscus maniculatus).

Total thyroxine was assessed by radioimmunoassay for 58 female deer mice of two subspecies (Peromyscus maniculatus bairdii and P. m. gracilis) and two color morphs (agouti and non-agouti). P. m. bairdii of both color-morphs had significantly higher mean thyroxine levels than P. m. gracilis. Non-agouti deer mice of both subspecies had significantly higher mean thyroxine levels than agouti deer mice. This is the first report of thyroid hormone differences associated with the non-agouti allele.     

Variations in the Coding Region of the Agouti Signaling Protein Gene Do Not Explain Agouti/Non-agouti Phenotypes in Macaques

Agouti is a common pigmentation phenotype in mammals including primates. Mutations in the agouti signaling protein gene (ASIP) are known to result in non-agouti black hairs in laboratory mice. It is still unclear whether sequence variation in ASIP is linked with the agouti/non-agouti phenotypes in macaques (Genus Macaca). To address this issue, we have determined and compared nucleotide sequences of protein coding region of ASIP in 18 macaque species and have identified 16 different sequences of the ASIP. Macaca nemestrina, which showed yellow agouti hairs, shared an identical amino acid sequence of ASIP with several non-agouti species. No sequence changes were found in functionally important sites of ASIP in the macaques showing non-agouti dark hair color. These results indicated that the variation in the protein coding region of ASIP did not explain the non-agouti dark coat color in the macaques. Upstream regulatory regions of ASIP and other genes participating in pigmentation system remain to be investigated for the hair color variation in the macaques.     

The genetics of coat colors in the mongolian gerbil (Meriones unguiculatus)

Genetic studies demonstrated three loci controlling coat colors in the Mongolian gerbil. F1 hybrids of white gerbils with red eyes and agouti gerbils with wild coat color had the agouti coat color. The segregating ratio of agouti and white in the F2 generation was 3:1. In the backcross (BC) generation (white x F1), the ratio of the agouti and white coat colors was 1:1. Next, inheritance of the agouti coat color was investigated. Matings between agouti and non-agouti (black) gerbils produced only agouti gerbils. In the F2 generation, the ratio of agouti to non-agouti (black) was 3:1. There was no distortion in the sex ratios within each coat color in the F1, F2 and BC generations. This indicated that the white coat color of gerbils is governed by an autosomal recessive gene which should be named the c allele of the c (albino) locus controlling pigmentation, and the agouti coat color is controlled by an autosomal dominant gene which might be named the A allele of the A (agouti) locus controlling pigmentation patterns in the hair. The occurrence of the black gerbil demonstrated clearly the existence of the b (brown) locus, and it clearly indicated that the coat colors of gerbils can basically be explained by a, b, and c loci as in mice and rats.     

Fecal glucocorticoids: a noninvasive method of measuring adrenal activity in wild and captive rodents

To determine the utility of fecal corticosteroid concentration as a measure of chronic stress under laboratory and field conditions, we biochemically and physiologically validated a radioimmunoassay for corticosteroids in three rodent species, house mice (Mus musculus), deer mice (Peromyscus maniculatus), and red-back voles (Clethrionomys gapperi). The biochemical validations demonstrated that the assay accurately and precisely measured corticosteroid concentration in the feces. The physiological validation indicated that the assay was sensitive enough to detect the stress associated with (a) brief handling and bleeding of animals, (b) chronic caloric restriction, (c) exposure to a novel environment, and (d) exposure to a novel cold environment. Our results suggest that fecal measurements reflect stress levels experienced by these animals approximately 6-12 h before defecation. Therefore, given a judicious trapping and trap-monitoring protocol, this assay has considerable utility for measuring the stress levels at which animals actually exist in the field.     

Genetic Instability at the Agouti Locus of the Mouse (Mus Musculus). I. Increased Reverse Mutation Frequency to the A(w) Allele in a/a Heterozygotes

We have compiled the reverse mutation rate data to the white bellied agouti (A(w)) allele in heterozygous A/a mice and shown it to be increased by a factor of at least 350 in comparison to the reverse mutation rate in homozygous a/a mice. Employing tightly linked flanking restriction fragment length polymorphism DNA markers, we have shown that reversion to A(w) is associated with crossing over in the vicinity of the agouti locus. The non-agouti (a) allele has been recently shown to contain an 11-kb insert within the first intron of the agouti gene. Together with our present results, these observations suggest possible mechanisms to explain the reversion events.     

Summer predation rates on ungulate prey by a large keystone predator: how many ungulates does a large predator kill?

Estimates of predation rates by large predators can provide valuable information on their potential impact on their ungulate prey populations. This is especially the case for pumas Puma concolor and its main prey, mule deer Odocoileus hemionus . However, only limited information on predation rates of pumas exist where mule deer are the only ungulate prey available. I used VHF telemetry data collected over 24-h monitoring sessions and once daily over consecutive days to derive two independent estimates of puma predation rates on mule deer where they were the only large prey available. For the 24-h data, I had 48 time blocks on female pumas with kittens, 43 blocks on females without kittens and 30 blocks on males. For the daily consecutive data, the average number of consecutive days followed was 51.5±4.2 days. There were data on five female pumas with kittens, five pregnant females and nine females without kittens. Predation rates over an average month of 30 days from the 24-h monitoring sessions were 2.0 mule deer per puma month for males (15.1 days per kill), 2.1 mule deer per puma month (14.3 days per kill) for females without kittens and 2.5 mule deer per puma month (12.0 days per kill) for pregnant females and females with kittens. For the consecutive daily data, females without kittens had an estimated predation rate of 2.1±0.14 mule deer per puma month (14.9±0.90 days per kill). Pregnant and females with kittens had predation rates of 2.7±0.18 and 2.6±0.21 mule deer per puma month, respectively (11.4±0.72 and 12.0±1.1 days per kill, respectively). Predation rates estimated in this study compared with those estimated by energetic demand for pumas in the study area but were lower than other field derived estimates. These data help increase our understanding of predation impacts of large predators on their prey.     

Environmental Enrichment Alters Splenic Immune Cell Composition and Enhances Secondary Influenza Vaccine Responses in Mice

Chronic stress has deleterious effects on immune function, which can lead to adverse health outcomes. However, studies investigating the impact of stress reduction interventions on immunity in clinical research have yielded divergent results, potentially stemming from differences in study design and genetic heterogeneity, among other clinical research challenges. To test the hypothesis that reducing glucocorticoid levels enhances certain immune functions, we administered influenza vaccine once (prime) or twice (boost) to mice housed in either standard control caging or environmental enrichment (EE) caging. We have shown that this approach reduces mouse corticosterone production. Compared with controls, EE mice had significantly lower levels of fecal corticosterone metabolites (FCMs) and increased splenic B and T lymphocyte numbers. Corticosterone levels were negatively associated with the numbers of CD19⁺ (r² = 0.43, p = 0.0017), CD4⁺ (r² = 0.28, p = 0.0154) and CD8⁺ cells (r² = 0.20, p = 0.0503). Vaccinated mice showed nonsignificant differences in immunoglobulin G (IgG) titer between caging groups, although EE mice tended to exhibit larger increases in titer from prime to boost than controls; the interaction between the caging group (control versus EE) and vaccine group (prime versus boost) showed a strong statistical trend (cage-group*vaccine-group, F = 4.27, p = 0.0555), suggesting that there may be distinct effects of EE caging on primary versus secondary IgG vaccine responses. Vaccine-stimulated splenocytes from boosted EE mice had a significantly greater frequency of interleukin 5 (IL-5)-secreting cells than boosted controls (mean difference 7.7, IL-5 spot-forming units/10⁶ splenocytes, 95% confidence interval 0.24–135.1, p = 0.0493) and showed a greater increase in the frequency of IL-5–secreting cells from prime to boost. Our results suggest that corticosterone reduction via EE caging was associated with enhanced secondary vaccine responses, but had little effect on primary responses in mice. These findings help identify differences in primary and secondary vaccine responses in relationship to stress mediators that may be relevant in clinical studies.     

The calm mouse: an animal model of stress reduction

Chronic stress is associated with negative health outcomes and is linked with neuroendocrine changes, deleterious effects on innate and adaptive immunity, and central nervous system neuropathology. Although stress management is commonly advocated clinically, there is insufficient mechanistic understanding of how decreasing stress affects disease pathogenesis. Therefore, we have developed a "calm mouse model" with caging enhancements designed to reduce murine stress. Male BALB/c mice were divided into four groups: control (Cntl), standard caging; calm (Calm), large caging to reduce animal density, a cardboard nest box for shelter, paper nesting material to promote innate nesting behavior, and a polycarbonate tube to mimic tunneling; control exercise (Cntl Ex), standard caging with a running wheel, known to reduce stress; and calm exercise (Calm Ex), calm caging with a running wheel. Calm, Cntl Ex and Calm Ex animals exhibited significantly less corticosterone production than Cntl animals. We also observed changes in spleen mass, and in vitro splenocyte studies demonstrated that Calm Ex animals had innate and adaptive immune responses that were more sensitive to acute handling stress than those in Cntl. Calm animals gained greater body mass than Cntl, although they had similar food intake, and we also observed changes in body composition, using magnetic resonance imaging. Together, our results suggest that the Calm mouse model represents a promising approach to studying the biological effects of stress reduction in the context of health and in conjunction with existing disease models.     

Effects of different photoperiods and handling stress on spawning and reproductive performance of pikeperch Sander lucioperca

The objective of this study was to control the reproductive cycle of pikeperch (Sander lucioperca) through determining the effects of different photoperiods and handling stress on the reproduction quality, timing and quality of spawning, fertilization, sex steroids, and cortisol concentrations. In this study, 72 pikeperch broodstocks with an average weight of 1367 ± 55.3 g were exposed to different photoperiods including constant light (24L:0D), constant darkness (0L:24D), and 12 h of light, 12 h of darkness (12L:12D) for 40 days. Half of the broodstocks of each photoperiod treatment were exposed to handling stress at a specific time of the day. Applying different photoperiods caused changes in the timing of broodstocks' spawning, so that fish under 24L:0D spawned earlier than those of other photoperiods, and stressed fish of the 0L:24D photoperiod had a delayed spawning compared to others. Also, the spawning of the broodstocks at different photoperiods which were exposed to handling stress was either delayed or did not occur at all. The highest and lowest spawnings were observed in the morning and at night, respectively. Fertilization percentage, number of eggs per gram, sex steroids including estradiol, progesterone, and testosterone, as well as cortisol and calcium concentrations did not show any significant difference in different photoperiods and handling stress. In stressed males of the 24L:0D photoperiod, there only was a significant decrease of testosterone concentration compared to the beginning of the experiment. Results indicated that the spawning performance of pikeperch broodstocks could be considerably stimulated using an effective photoperiod. Similarly, pikeperch broodstocks in culture systems are usually affected by handling stress, and this stress could lead to a poor reproductive performance and inhibition of spawning.     

Psychological Stress on Female Mice Diminishes the Developmental Potential of Oocytes: A Study Using the Predatory Stress Model

Although the predatory stress experimental protocol is considered more psychological than the restraint protocol, it has rarely been used to study the effect of psychological stress on reproduction. Few studies exist on the direct effect of psychological stress to a female on developmental competence of her oocytes, and the direct effect of predatory maternal stress on oocytes has not been reported. In this study, a predatory stress system was first established for mice with cats as predators. Beginning 24 h after injection of equine chorionic gonadotropin, female mice were subjected to predatory stress for 24 h. Evaluation of mouse responses showed that the predatory stress system that we established increased anxiety-like behaviors and plasma cortisol concentrations significantly and continuously while not affecting food and water intake of the mice. In vitro experiments showed that whereas oocyte maturation and Sr²⁺ activation or fertilization were unaffected by maternal predatory stress, rate of blastocyst formation and number of cells per blastocyst decreased significantly in stressed mice compared to non-stressed controls. In vivo embryo development indicated that both the number of blastocysts recovered per donor mouse and the average number of young per recipient after embryo transfer of blastocysts with similar cell counts were significantly lower in stressed than in unstressed donor mice. It is concluded that the predatory stress system we established was both effective and durative to induce mouse stress responses. Furthermore, predatory stress applied during the oocyte pre-maturation stage significantly impaired oocyte developmental potential while exerting no measurable impact on nuclear maturation, suggesting that cytoplasmic maturation of mouse oocytes was more vulnerable to maternal stress than nuclear maturation.     

Applying modern analogs to understand the pollen content of coprolites

Most scientists working with coprolites from archaeological contexts assume that human fecal specimens reflect the mixing of the pollen ingested during the period in which the contribution to the coprolite, 19 to 37.5 h, was ingested, that the amount of pollen in a fecal sample directly reflects the amount of pollen originally ingested during that interval, and that differences between the amounts of pollen in different fecal specimens reflect differences in the quantities of pollen ingested at different times. These assumptions were tested and found wanting in an experiment in which two persons sequentially ate separate quantities of 15 pollen types in meals over a four-day interval. The pollen was retrieved and analyzed from feces produced during those four days and five days of subsequent fecal production. Pollen ingested first appeared in relatively small amounts, usually the day after it was ingested. Its concentration per gram of sample then increased rapidly and remained high over a one to three day interval relative to the amounts in previous and subsequent fecal specimens deposited. When pollen concentrations declined some pollen was retained in the gastrointestinal system and much lower concentrations per gram of sample of each type continued to appear in fecal samples for several days. These relatively low pollen concentrations appeared in fecal samples approximately twice as often as did higher concentrations. Our results indicate that comparatively high pollen concentrations can be used to determine that a given pollen type was ingested, but comparisons between pollen concentrations of the same pollen type in different fecal specimens or between different pollen types in the same fecal specimen, cannot be used to determine whether different amounts of pollen were ingested, or what was the relative amount of each ingested. Because pollen concentrations per gram of sample varied widely with time since ingestion, percentages of given pollen types did not occur in predictable patterns and could actually increase as the concentration of the pollen type decreases. Hence, percentages should not be used in coprolite pollen analysis. The experimental results also suggest that variations in the pollen content of different portions of a coprolite are meaningful only in terms of the overall pattern of a sequential group of coprolites.     

Influence of experimental context on the development of anhedonia in male mice exposed to chronic social stress

Consumption of 1% sucrose solution supplemented with 0.2% vanillin was studied in two experimental contexts in male mice living under chronic social stress induced by daily experience of defeats in agonistic interactions and leading to development of depression. In the first experiment, vanillin sucrose solution was made available as an option along with water during 10 days for mice living in group home cages. Then the mice were subjected to repeated social defeat stress and during exposure to stress they were provided with both vanillin sucrose solution and water using a free two-bottle choice paradigm. In the other experiment, vanillin sucrose solution was first offered to mice after 8 days of exposure to social defeat stress. Males familiar with vanillin sucrose solution showed vanillin sucrose preference while experiencing defeat stress: consumption of vanillin sucrose solution was about 70% of total liquid consumption. However, the consumption of vanillin sucrose solution per gram of body weight in mice exposed to social stress during 20 days was significantly lower than in control males. In the second experiment, males after 8 days of social defeat stress were found to consume significantly less vanillin sucrose solution as compared to control males. On average, during two weeks of measurements, vanillin sucrose solution intake was less than 20% of total liquid consumption in males. Consumption per gram of body weight also appeared to be significantly lower than in control group. The influence of experimental context on the development of anhedonia measured as a reduction of sucrose solution intake by chronically stressed male mice is discussed.     

Testosterone production in mice lacking inducible nitric oxide synthase expression is sensitive to restraint stress

Immobilization stress (IMO) induces a rapid increase in glucocorticoid secretion [in rodents, corticosterone CORT)] and this is associated with decreased circulating testosterone (T) levels. Nitric oxide (NO), a reactive free radical and neurotransmitter, has been reported to be produced at higher rates in tissues such as brain during stress. The biosynthesis of T is also known to be dramatically suppressed by NO. Specifically, the inducible isoform of nitric oxide synthase (iNOS) was directly implicated in this suppression. To assess the respective roles of CORT and NO in stress-mediated inhibition of T production, adult wild-type (WT) and inducible nitric oxide synthase knockout (iNOS(-/-)) male mice were evaluated. Animals of each genotype were assigned to either basal control or 3-h IMO groups. Basal plasma and testicular T levels were equivalent in both genotypes, whereas testicular weights of mutant mice were significantly higher compared with WT animals. Exposure to 3-h IMO increased plasma CORT and decreased T concentrations in mice of both genotypes. Testicular T levels were also affected by stress in WT and mutant males, being sharply reduced in both genotypes. However, the concentrations of nitrite and nitrate, the stable metabolites of NO measured in testicular extracts, did not differ between control and stressed WT and iNOS(-/-) mice. These results support the hypothesis that CORT, but not NO, is a plausible candidate to mediate rapid stress-induced suppression of Leydig cell steroidogenesis.     

Role of oviposition preference in an invasive crambid impacting two graminaceous host crops

Oviposition preference studies of the Mexican rice borer, Eoreuma loftini (Dyar), on sugarcane, Saccharum spp., and rice, Oryza sativa L., showed that drought stressed sugarcane was 1.8-fold more attractive based on egg masses/plant than well watered sugarcane. The E. loftini susceptible sugarcane cultivar LCP 85-384 was 1.6-fold more attractive than HoCP 85-845 based on numbers of eggs per egg mass. Egg masses were 9.2-fold more abundant and 2.3-fold larger on sugarcane than on rice. Rice, however, was preferred to sugarcane on a plant biomass basis. Oviposition on sugarcane occurred exclusively on dry leaf material, which increased under drought stress. Egg masses per plant increased on drought stressed sugarcane and were correlated with several foliar free amino acids essential for insect growth and development. The more resistant (based on injury) but more attractive (based on oviposition) rice cultivar XL8 had higher levels of several free amino acids than the susceptible cultivar Cocodrie. The association of host plant characteristics to oviposition preference is discussed. Projected oviposition patterns relative to sugarcane and rice production areas were estimated for Texas and Louisiana based on the availability of each host in different regions of each state. These results suggest that, where sugarcane and rice co-occur, the majority of eggs would be found on sugarcane early in the season, because of this crop's substantially greater biomass compared with rice. Abundance later in the season would also favor sugarcane; however, the abundance on rice would be greater than expected solely based on host availability, largely because of the greater preference per gram of rice plant dry weight.     

Relationship between Ethylene and Spermidine in the Leaves of Glycyrrhiza uralensis Seedlings under Root Osmotic Stress1

The correlation between ethylene (ETH) production and spermidine (Spd) content was studied in the leaves of Glycyrrhiza uralensis seedlings under root osmotic stress. After 4-h root osmotic stress, Spd inhibited ETH production significantly while ETH had no significant influence on Spd content in leaves. After 24-h root osmotic stress, ETH production and Spd content showed significant negative correlations in leaves, and the negative correlations were still significant even if the common precursor S-adenosylmethionine (SAM) was abundant. This result suggested that the significant negative correlations were not mainly caused by the competition for the common precursor SAM. The results also showed that, after 24-h root osmotic stress, ETH enhanced while Spd reduced the production rate of H₂O₂ and markedly in leaves; furthermore, exogenous H₂O₂ accelerated the increase in ETH production and the decrease in Spd content caused by deep stress. So it could be concluded that reactive oxygen species played important roles in the significant negative correlations in the deeply stressed leaves of G. uralensis seedlings.     

Platinum coat color locus in the deer mouse

Platinum coat color in the deer mouse, Peromyscus maniculatus, is an autosomal recessive trait marking a locus, pt, distinct from silver (si), albino (c), blonde (bl), brown (b), and agouti (a). Platinum deer mice are conspicuously pale, with light ears and tail stripe. The pewter trait is allelic with and phenotypically identical to platinum, and represents an independent recurrence of this mutant. The rate of recoveries of coat color mutations from wild deer mice is consistent with available data for recurring mutation rates balanced by strong selection against the recessive phenotype.     

Altered adrenal gland cholesterol metabolism in the apoE-deficient mouse

Previous studies suggest the hypothesis that apoE produced by adrenocortical cells modulates cellular cholesterol metabolism to enhance the storage of esterified cholesterol (EC) at the expense of cholesterol delivery to the steroidogenic pathway. In the present study, parameters of adrenal cholesterol metabolism and corticosteroid production were examined in wild type and apoE-deficient (apoe(-/-)) mice. Adrenal gland EC content and the EC/free cholesterol (FC) ratio in mice stressed by adrenocorticotropin (ACTH) treatment or saline injection were reduced in apoe(-/-) compared to apoe(+/+) mice. Relative to apoe(+/+) mice, apoE deficiency also resulted in increased levels of plasma corticosterone in the basal state, in response to acute or long-term ACTH treatment, and after a swim-induced neuroendocrine-directed stress test. Measurements of adrenal gland scavenger receptor class B, type I (SR-BI), LDL receptor, and LDL receptor related protein (LRP) levels and the activities of ACAT or HMG-CoA reductase showed no difference between genotypes. Apoe(-/-) and apoe(+/+) mice showed similar quantitative increases in LDL receptors, SR-BI, adrenal weight gain, and ACAT activities in response to ACTH, and both genotypes had similar basal plasma ACTH concentrations. These results suggest that the effects of apoE deficiency reflect events at the level of the adrenal gland and are specific to changes in cholesterol accumulation and corticosterone production. Further, these findings support the hypothesis that apoE acts to enhance adrenocortical EC accumulation and diminish corticosterone production.     

Microtransferrinuria and microalbuminuria. II. In the rat

We studied albumin and transferrin excretion in the normal and diabetic rat: (1) The rat secretes small concentrations of albumin and transferrin in the urine. (2) The secretion depends on the strain and was highest in the Kyoto spontaneously hypertensive rat. (3) The secretion of these two proteins in the rat is quite dependent on age and sex. The level increases dramatically with age. The secretion is much higher in the male compared to the female. This difference is observed after puberty. The changes in transferrin relative to those in albumin are much higher. (4) In streptozotocin-induced diabetes, the concentration of albumin and transferrin expressed as milligrams per liter decreases; however, the output/24 h or per gram creatinine is increased with a greater increase in transferrin output relative to that of albumin. The similarities and differences between excretion of these two proteins in the human and the rat as well as their importance are discussed.     

Verification of the color genotype based on morphological analysis of fur pigmentation of brown and black water voles Arvicola terrestris L.]

Qualitative and quantitative analysis of fur pigmentation in brown and black water voles (Arvicola terrestris L.) was performed. Morphology of pigment granules, their distribution along the hair layers and histology of hair bulbs were studied. Morphological data and the results of the analysis of segregation in the progeny, when brown voles were self-crossed and crossed with black ones, led to conclusion that fur colour of water voles is mediated by genes from the agouti series, precisely, black colour is determined by the extreme non-agouti allele (aeae genotype), and in brown voles which are homo- or heterozygous the colour is determined by the agouti allele (AA or Aae genotypes).