Vasopressin and aggression in cross-fostered California mice (Peromyscus californicus) and white-footed mice (Peromyscus leucopus)

To examine how developmental experiences alter neural pathways associated with adult social behavior, we cross-fostered pups between the more aggressive and monogamous California mouse (Peromyscus californicus) and the less aggressive and polygamous white-footed mouse (P. leucopus). Cross-fostered males became more like their foster parents when tested as adults. Male white-footed mice became more aggressive only in an aggression test in a neutral arena, whereas the territorial California mice became less aggressive in resident-intruder aggression test, as measured by attack latency. Only the species that displayed a change in resident-intruder aggression showed a change in arginine vasopressin (AVP) levels: cross-fostered California mice had significantly lower levels of AVP-immunoreactive (AVP-ir) staining than controls in the bed nucleus of the stria terminalis (BNST) and the supraoptic nucleus (SON) and a nonsignificant trend toward lower levels in the medial amygdala (MA). Neither species showed changes in AVP-ir staining in a control area, the paraventricular nucleus (PVN). The changes in AVP-ir staining in the BNST and SON may not be caused by stress because cross-fostering was not associated with changes in adult plasma concentrations of two steroid hormones, corticosterone and testosterone, that have been associated with stress-related alterations in AVP pathways. These results suggest that manipulating the early parental environment can directly alter both a neurotransmitter system and species-typical patterns of social behavior, but that these effects may vary between species and under different social contexts.     

Bioenergetic benefits of huddling by deer mice (Peromyscus maniculatus)

Both short photoperiod and communal social living conserve metabolic energy by deer mice held in thermal neutral ambient temperatures. Initial socialization was energetically more costly than solitary living, but huddling behaviors reduced thermal conductance and mass specific metabolic rate by 30% within 5 days. While short photoperiod reduced metabolic energy expenditure by decreasing thermoregulatory demand, huddling mediated behavioral conservation was achieved with hyperthermic core temperatures.     

Developmental plasticity in aerobic performance in deer mice (Peromyscus maniculatus)

While several studies have examined the abiotic effects of altitude (low ambient temperatures and hypoxia) on the aerobic performance of small mammals, few have explored the effects of development and maturation at different altitudes on aerobic performance as adults. We examined the basal metabolism and aerobic performance of deer mice (Peromyscus maniculatus) under four different developmental and testing regimes: (1) reared (gestation through weaning) and tested at high altitude; (2) reared and tested at low altitude; (3) reared at low altitude and tested at high altitude after acclimation; and (4) reared at low altitude and tested in hypoxia without acclimation. We found that mice that developed and were tested at low altitudes had a higher aerobic capacity (both aerobic performance and basal metabolic rate) than those that developed, or were acclimated as adults, at high altitudes. In addition, we found that mice that developed at high altitude did not have a higher aerobic capacity than those that developed at low altitude and were acclimated to high altitude as adults. Both groups tested at high altitudes had higher hematocrits (% red blood cells) and hemoglobin than mice tested at low altitudes. Surprisingly, mice acclimated to low altitudes and given an instantaneous exposure to hypoxia did not suffer a depression in aerobic performance.     

Fecal corticosteroids in agouti and non-agouti deer mice (Peromyscus maniculatus)

Total and per gram fecal corticosteroid concentrations were determined for agouti and non-agouti deer mice (Peromyscus maniculatus gracilis) over 24 h under normal caging conditions and after exposure to the stress of novel caging. Per gram corticosteroid concentrations, fecal output, and 24-h corticosteroid production were greater in stressed compared with unstressed deer mice of both color morphs, whereas stressed agoutis had a greater increase in per gram corticosteroid concentrations when compared with non-agoutis. However, due to increased fecal output, stressed non-agouti deer mice had greater 24-h corticosteroid production. Thus, agouti and non-agouti deer mice differ in their hormonal reaction to stress. This is the first demonstration of corticosteroid differences associated with the agouti locus.     

Ultrasonic vocalizations and ontogenetic development in California mice (Peromyscus californicus)

This experiment investigated the time-course of behavioral and physical development in 18 male and 18 female pups from 18 litters of California mice. From 1 to 23 days of age animals were observed for a 6-min observation period every other day, followed by a neurobehavioral testing session. Ultrasonic vocalizations (UVs), coordinated movements, neuromotor indicators and physical parameters were measured. The production of UVs peaked during the first week of development, remained stable from 9 to 15 days of age, and decreased abruptly thereafter. Females vocalized more than males during the first 9 days after birth, although this difference only was statistically significant on the third day. There was an inverse relationship between the emission of UVs and coordinated movements. The frequency of UVs dropped significantly 1 day after the pups had displayed surface righting, were able to hear, and had begun opening their eyes compared with 1 day before. In conclusion, it appears that there is a sex difference in the emission of UVs, which is more intense at the beginning of development. Furthermore, the ontogenetic patterns of changes in the emission of UVs, by both males and females in California mice, are inversely related to the development of coordinated movements, such as rearing, locomotion and self-grooming.     

Paternal experience and stress responses in California mice (Peromyscus californicus)

Paternal behavior greatly affects the survival, social development, and cognitive development of infants. Nevertheless, little research has been done to assess how paternal experience modifies the behavioral characteristics of fathers, including fear and stress responses to a novel environment. We investigated long-term behavioral and physiologic effects of parental experience in mice (Peromyscus californicus) and how this response activates the hypothalamic-pituitary-adrenal axis (as measured by corticosterone and dehydroepiandrosterone [DHEA] levels) and interacts with anxiety-related behaviors. Three groups of adult males were tested--fathers exposed to pups, virgins exposed to pups, and virgins never exposed to pups--in 2 environments designed to elicit anxiety response: an open field with a novel object placed in the center and a closed cage containing a sample of a component of fox feces. Behavioral responses were measured by using traditional methods (duration and frequency) and behavioral-chain sequences. Results indicated that paternal experience significantly modifies a male mouse's behavioral and physiologic responses to stress-provoking stimuli. Compared with inexperienced male mice, experienced male mice had a significant decrease in the occurrence of incomplete behavioral chains during the exposure to the novel object, an index of reduced stress. Further, even moderate pup exposure induced behavioral modifications in virgin male mice. These behavioral responses were correlated with changes in corticosterone and DHEA levels. Together, these data provide evidence that interactions between male mice and offspring may have mutually beneficial long-term behavioral and physiologic effects.     

Frequent Harderian gland adenocarcinomas in inbred white-footed mice (Peromyscus leucopus)

In 1997, three lines of inbred Peromyscus leucopus--GS109A, GS16A1, and GS16B--were acquired by the Peromyscus Genetic Stock Center. Since then, records have been kept on tumors detected by visible inspection of live animals. The inbred lines GS109A and GS16A1 presented tumors with frequencies substantially higher than that of the other inbred line or of random-bred P. leucopus stock. The average age of detection was 456 +/- 75 days (n = 24) for GS109A and 568 +/- 168 days (n = 12) for GS16A1 respectively. Surprisingly, the majority of the tumors (23 of 24 for GS109A and 8 of 12 for GS16A1) appeared to be Harderian gland lesions. During the same time period only a single tumor, a fibrosarcoma, was noted in the other inbred strain (GS16B), and one Harderian gland tumor was detected in the random bred stock. On the basis of the number of animals born to each group, tumor frequencies were approximately 22.7%, 8.3%, 0.67%, and 0.07%, for GS109A, GS16A1, GS16B, and randombred P. leucopus stock, respectively. The periocular tumors appeared to be highly malignant, with elevated mitotic indices, marked anaplasia, and metastases to regional lymph nodes and lungs. The tumors were readily transplantable to other animals of the same line. Among various other species, malignant Harderian gland tumors are relatively rare.     

Modoc-like virus isolated from wild deer mice (Peromyscus maniculatus) in Alberta

Small mammals were trapped in northeastern Alberta, Canada during 1976. Blood samples from these animals were tested for virus by inoculation of suckling mice. Blood clots from two deer mice yielded isolates of the same virus. The virus was related antigenically to a number of flaviviruses which have been isolated from mammals in Central America and North America and was related most closely to Modoc virus. Physical, chemical, and biological properties of the virus were similar also to those of Modoc virus. It did not produce illness or death in deer mice inoculated in the laboratory. Neutralization tests indicated that 1/38 (3%) red squirrels (Tamiasciurus hudsonicus), 3/35 (9%) least chipmunks (Eutamius minimus), 13/109 (12%) deer mice, and 3/50 (6%) humans were infected naturally. This is the first reported evidence of infection of red squirrels and chipmunks with a Modoc-like virus. These data extend the range of Modoc-like viruses northward by 1,500 km and comprise the first isolate from mammals in the boreal forest of Canada.     

Effects of heavy metals on immunocompetence of white-footed mice (Peromyscus leucopus)

Continued human population growth and industrialization result in increased contamination of wildlife habitats. Effects of such habitat deterioration on the well-being of natural populations are unclear. Exposure to contaminants may impair immunocompetence, thereby increasing disease susceptibility. The mammalian immune system is important in maintaining health and in its sensitivity to toxins. In our study conducted from May 1999 through May 2001, we examined assays of immnnoompetence in the white-footed mouse (Peromyscus leucopus) that inhabited reference sites and sites significantly contaminated with mixtures of heavy metals. We estimated potential exposure and uptake of heavy metals by measuring the level of each contaminant in representative soil and tissue samples. Intraindividual variation across mice, but not sex, explained a large portion of the overall variance in immune response, and spleen weight was significanltly afflicted by mouse age. We found no evidence that residence on contaminated sites had any effect on immunopathology and humoral immunity as measured in our study. We suggest that field and laboratory studies in ecotoxicology provide estimates of exposure to contaminants (i.e., tissue analyses) to establish a database suitable to clarify the dose-response relationship between contaminants and target systems.     

Estrogen-dependent modifications to hippocampal plasticity in paternal California mice (Peromyscus californicus)

In many biparental species, mothers and fathers experience similar modifications to circulating hormones. With these modifications come alterations in neural structure and function suggesting that neuroendocrine mechanisms may underlie postpartum plasticity in both males and females. In the biparental California mouse (Peromyscus californicus), adult neurogenesis is maintained and anxiety-like behavior is attenuated in fathers during the mid-postpartum period. Given a causal relationship between estrogen and regulation of both adult neurogenesis and anxiety, we aimed to elucidate the role of estrogen-dependent mechanisms in paternal experience-related modifications to hippocampal neuroplasticity in California mice. In Experiment 1, hippocampal estrogen receptor beta (ERβ) mRNA expression, along with circulating estradiol concentrations, were determined throughout the postpartum period. An upregulation in ERβ expression was observed in postnatal day 16 males compared to virgins. Additionally, a rise in circulating estradiol concentrations was detected on postnatal day 2 compared to virgins; levels began to decline toward virgin levels on postnatal day 16 and postnatal day 30. In Experiment 2, we determined the role of estrogen-dependent mechanisms in adult neurogenesis and anxiety-like behavior by treating virgin and paternal males with saline or the selective estrogen receptor modulator, tamoxifen (TMX), during the time of axon extension (i.e., one week after bromodeoxyuridine injection). While TMX failed to alter elevated plus maze performance, TMX treatment inhibited survival of adult born neurons but only in paternal mice. These findings highlight the potential for estrogen-dependent pathways to mediate hippocampal adult neurogenesis in paternal mice.     

Responses to lactation and cold exposure by deer mice (Peromyscus maniculatus)

Recently, much interest has been expressed in understanding how animals use phenotypic plasticity of tissue size and function to meet increased metabolic demands. We set out to learn (i) whether female deer mice (Peromyscus maniculatus) given lactation (two to seven pups per litter), cold (5 degrees C), or cold plus lactation as energy demands display phenotypic plasticity in organ size and function; (ii) whether that plasticity is similar to laboratory mice given the same demands; and (iii) whether lactational performance in deer mice is derived from limits on central or peripheral organs. We found that deer mice responded to lactation by increasing digestible food intake and increasing the masses of the stomach, small intestine, cecum and liver, and the length of the small intestine. Heart mass was lower in lactating than in nonlactating mice. Cold exposure also caused increases in digestible food intake and increases in the masses of the small intestine, kidney, and heart. We conclude that deer mice display organ tissue plasticity in response to both lactation and cold exposure in a similar manner to laboratory mice. We also conclude that deer mice are not limited by central processing organs because they are able to increase digestive organ size continuously with increased energetic demands.     

Population delimitation across contrasting evolutionary clines in deer mice (Peromyscus maniculatus)

Despite current interest in population genetics, a concrete definition of a "population" remains elusive. Multiple ecologically and evolutionarily based definitions of population are in current use, which focus, respectively, on demographic and genetic interactions. Accurate population delimitation is crucial for not only evolutionary and ecological population biology, but also for conservation of threatened populations. Along the Pacific Coast of North America, two contrasting patterns of geographic variation in deer mice (Peromyscus maniculatus) converge within the state of Oregon. Populations of these mice diverge morphologically across an east-west axis, and they diverge in mitochondrial DNA haplotypes across a north-south axis. In this study, we investigate these geographically contrasting patterns of differentiation in the context of ecological and evolutionary definitions (paradigms) of populations. We investigate these patterns using a new and geographically expansive sample that integrates data on morphology, mitochondrial DNA, and nuclear DNA. We found no evidence of nuclear genetic differentiation between the morphologically and mitochondrially distinct populations, thus indicating the occurrence of gene flow across Oregon. Under the evolutionary paradigm, Oregon populations can be considered a single population, whereas morphological and mitochondrial differentiations do not indicate distinct populations. In contrast, under the ecological paradigm morphological differentiation indicates distinct populations based on the low likelihood of demographic interactions between geographically distant individuals. The two sympatric but mitochondrially distinct haplogroups form a single population under the ecological paradigm. Hence, we find that the difference between evolutionary and ecological paradigms is the time-scale of interest, and we believe that the more chronologically inclusive evolutionary paradigm may be preferable except in cases where only a single generation is of interest.     

The biology and methodology of assisted reproduction in deer mice (Peromyscus maniculatus)

Although laboratory-reared species of the genus Peromyscus—including deer mice—are used as model animals in a wide range of research, routine manipulation of Peromyscus embryogenesis and reproduction has been lagging. The objective of the present study was to optimize conditions for oocyte and/or embryo retrieval and for in vitro culturing. On average, 6.4 oocytes per mouse were recovered when two doses of 15 IU of pregnant mare serum gonadotropin (PMSG) were given 24 h apart, followed by 15 IU of hCG 48 h later. Following this hormone priming, females mated overnight with a fertile male yielded an average of 9.1 two-cell stage embryos. Although two-cell stage embryos developed to 8-cell stage in Potassium Simplex Optimized Medium (KSOM; Millipore-Chemicon, Billerica, MA, USA) in vitro, but not further, embryos recovered at the 8- to 16-cell stages developed into fully expanded blastocysts when cultured in M16 media in vitro. These blastocysts had full potential to develop into late stage fetuses and possibly into live pups. As a result of the present work, all stages of Peromyscus preimplantation development are now obtainable in numbers sufficient for molecular or other analyses. These advances provide the opportunity for routine studies involving embryo transfer (e.g., chimeras, transgenics), and preservation of genetic lines by cryopreservation.     

Annual cyclicity of gall stone prevalence in deer mice (Peromyscus maniculatus gambelii)

The prevalence of gall stones (100% cholesterol) in a deer mouse (Peromyscus maniculatus) population located in Siskiyou County, California (USA) was studied on a monthly basis from February 1985 through January 1988. During each year we documented a pronounced annual cyclicity with peak prevalence (31 to 53%) during the winter and low prevalence (2 to 3%) during late summer. Gall stone prevalence was not related to sex or age of the animal. The earliest onset of gall stone production and the maximum prevalence achieved were associated with the greatest abundance of deer mice; lower levels of population abundance were associated with later onset of gall stone production and lower peak prevalences. This association of gall stone prevalence with both season and population abundance levels suggests that the causative factor(s) is/are related to the quality and availability of the diet.     

Experimental infection and horizontal transmission of Modoc virus in deer mice (Peromyscus maniculatus)

Deer mice (Peromyscus maniculatus) were inoculated with a sublethal dose of a field strain of Modoc virus to determine patterns of viral persistence, shedding, and transmission. Blood, serum, urine, fecal, and oral swab samples were collected at selected intervals until 63 days postinoculation (PI) after which lung, liver, spleen, kidney, and salivary glands were explanted. Viral assays were conducted by intracranial inoculations of suckling mice and antibody titers were determined by the micro-complement-fixation test. Viremias lasted for up to 4 days PI. Antibody titers were present by day 8 PI, peaked at day 13-20 PI, and persisted until day 63 PI. There was no evidence of viral shedding in urine, fecal, or oral swab samples. Virus was detected in explanted lungs only. In a separate experiment, deer mice were inoculated with virus and lungs were removed from five mice per wk for 10 wk. Indirect fluorescent antibody (IFA) techniques were used to determine the location of virus in lung tissue and to examine fixed tissue for lesions. IFA showed virus in lung parenchymal cells beginning 42 days PI and persisting at least 70 days PI. No histopathologic changes were seen. Horizontal transmission of the virus was studied by placing uninoculated mice with inoculated mice for 42 days and determining if the test animals developed antibodies or had virus in their lungs. Fifty-percent of the uninoculated mice developed antibody. One of these animals had virus in its lungs. Therefore, Modoc virus may be transmitted by direct contact.     

Bone loss is a physiological cost of reproduction in white-footed mice (Peromyscus leucopus)

Investment in offspring production often requires the mobilization of endogenous resources, a strategy that may negatively impact maternal condition. In mammals, skeletal ossification in growing offspring requires a large investment of calcium by mothers, and bone loss has been described in several species as a means of supporting this demand. Although bone loss can have adverse effects on the mother, its potential role in a reproductive trade-off has not been addressed. Using white-footed mice (Peromyscus leucopus), we tested the effect of dietary calcium availability on maternal skeletal condition during reproduction to assess if calcium availability drives a trade-off between maternal skeletal condition and offspring production. We provided mice with a low-calcium or standard diet and monitored reproductive output along with changes in bone mineral density and bone resorption (via serum concentrations of pyridinoline cross-links) throughout reproduction. Reproductive performance was not impaired by low calcium intake. Reproductive females on the low-calcium diet showed a significant reduction in bone mineral density relative to reproductive females consuming the standard diet and non-reproductive mice consuming the low-calcium diet, but no difference in bone resorptive activity. Our results suggest that when dietary calcium is limited white-footed mice reproduce at the expense of their skeletal condition, and may do so by limiting bone mineral accretion relative to resorption.