White-footed Mouse Populations Research Articles

Genetic variation of Borreliella burgdorferi in Fairfax County, Virginia, targeting the OspC gene in white-footed mice.

Outer surface protein C (OspC) is a commonly used marker in population studies of Borreliella to differentiate types and establish evolution over time. Investigating the ospC genetic types of Borreliella burgdorferi across multiple organ tissues of white-footed mice has the potential to contribute to our understanding of Lyme disease and the wide spectrum of clinical presentation associated with infection. In this study, five unique tissue types were sampled from 90 mice and screened for B. burgdorferi infections. This initial screening revealed a 63% overall B. burgdorferi infection rate in the mice collected (57/90). A total of 163 tissues (30.4%) tested positive for B. burgdorferi infections and when mapped to Borreliella types, 143,894 of the initial 322,480 reads mapped to 10 of the reference sequences in the ospC strain library constructed for this study at a 97% MOI. Two tissue types, the ear and the tongue, each accounted for 90% of the observed Borreliella sequence diversity in the tissue samples surveyed. The largest amount of variation was observed in an individual ear tissue sample with six ospC sequence types, which is equivalent to 60% of the observed variation seen across all tested specimens, with statistically significant associations observed between tissue type and detected Borreliella. There is strong evidence for genetic variability in B. burgdorferi within local white-footed mouse populations and even within individual hosts by tissue type. These findings may shed light on drivers of infection sequalae in specific tissues in humans and highlights the need for expanded surveillance on the epigenetics of B. burgdorferi across reservoirs, ticks, and infected patients.

Demography of botfly (Cuterebra fontinella) parasitism in white-footed mice (Peromyscus leucopus) in Kansas

Abstract Botflies (Diptera: Cuterebra sp.) are widely distributed and seasonally abundant parasites of small mammals in North America. To assess their effect on rodent survival, we studied the demography of botfly parasitism on small mammals in northeastern Kansas during 1995–1999. Additional comparisons on botflies parasitizing small mammals were made to a nearby old-field grid where mark–recapture studies continued from 1975 to 2003. White-footed mice, Peromyscus leucopus, were infected by botfly larvae (Cuterebra fontinella) each year during our study. The period of infection for P. leucopus was June–December, with the highest proportion of mice infected in July and August. A smaller, second peak of infection in October and November indicated that C. fontinella reproduced bimodally at this latitude with distinct summer and autumn population peaks. During the botfly seasons of the 5 years, 140 (23.3%) of 602 P. leucopus captured were infected. The percentages of individuals infected during 1995, 1996, 1997, and 1999 were not significantly different (22.4%, 17.3%, 17.1%, and 22.7%, respectively). However, in 1998, 32.4% individuals were infected with a higher incidence of multiple infections and a greater number of botfly larvae per host. Nearly half (47%) of the infected individuals caught in 1998 had infections in both summer and autumn, compared with 25% during the 1995–1997 seasons. In 1999, 63% of infected mice had at least a 2nd infection during the autumn. For all 5 years, 82%, 14%, and 4% of the infected mice had one, two, or three botflies, respectively. In 1998, 24% of the infected mice carried two or more botflies at one time, compared to 14% from 1995–1997 and 11% in 1999. Overall 94% of the infected mice were adults, with no differences between sexes. The variation seen in the prevalence of botfly parasitism of white-footed mice correlates to yearly weather fluctuations. Other demographic aspects of the infected mice, such as sex ratio, age, and longevity of those infected, did not change year to year. Our data suggest that infection with Cuterebra has little negative or positive impact upon populations of white-footed mice.

Effects of physical impairments on fitness correlates of the white-footed mouse, Peromyscus leucopus.

Physical impairments are widely assumed to reduce the viability of individual animals, but their impacts on individuals within natural populations of vertebrates are rarely quantified. By monitoring wild populations of white-footed mice over 26 years, we assessed whether missing or deformed limbs, tail or eyes influenced the survival, body mass, movement and ectoparasite burden of their bearers. Of the 27 244 individuals monitored, 543 (2%) had visible physical impairments. Persistence times (survival) were similar between mice with and without impairments. Mice with eye and tail impairments had 5% and 6% greater mass, respectively, than unimpaired mice. Mice with tail impairments had larger home ranges than did unimpaired mice. Burdens of black-legged ticks (Ixodes scapularis) were higher among mice with tail and limb impairments while burdens of bot fly larvae (Cuterebra) were higher among mice with cataracts compared to mice without impairments. Our findings do not support the presupposition that physical impairments reduce viability in their bearers and are inconsistent with the devaluation of impaired individuals that pervaded early thinking in evolutionary biology.

Evolution and developmental plasticity of lung structure in high-altitude deer mice.

Hypoxia at high altitudes can constrain the ability of endotherms to maintain sufficient rates of pulmonary O2 transport to support exercise and thermogenesis. Hypoxia can also impede lung development during early post-natal life in some mammals, and could thus accentuate constraints on O2 transport at high altitude. We examined how these challenges are overcome in deer mice (Peromyscus maniculatus) native to high altitude. Lung structure was examined in highland and lowland populations of deer mice and lowland populations of white-footed mice (P. leucopus; a congener restricted to low altitude) that were bred in captivity. Among mice that were born and raised to adulthood in normoxia, highland deer mice had higher alveolar surface density and more densely packed alveoli. The increased alveolar surface density in highlanders became fully apparent at juvenile life stages at post-natal day 30 (P30), after the early developmental period of intense alveolus formation before P21. Alveolar surface density was maintained in highlanders that were conceived, born, and raised in hypoxia (~ 12kPa O2), suggesting that lung development was not impaired by post-natal hypoxia as it is in many other lowland mammals. However, developmental hypoxia increased lung volume and thus augmented total alveolar surface area from P14. Overall, our findings suggest that evolutionary adaptation and developmental plasticity lead to changes in lung morphology that should improve pulmonary O2 uptake in deer mice native to high altitude.

Signatures of positive selection and local adaptation to urbanization in white-footed mice (Peromyscus leucopus).

Urbanization significantly alters natural ecosystems and has accelerated globally. Urban wildlife populations are often highly fragmented by human infrastructure, and isolated populations may adapt in response to local urban pressures. However, relatively few studies have identified genomic signatures of adaptation in urban animals. We used a landscape genomic approach to examine signatures of selection in urban populations of white-footed mice (Peromyscus leucopus) in New York City. We analysed 154,770 SNPs identified from transcriptome data from 48 P.leucopus individuals from three urban and three rural populations and used outlier tests to identify evidence of urban adaptation. We accounted for demography by simulating a neutral SNP data set under an inferred demographic history as a null model for outlier analysis. We also tested whether candidate genes were associated with environmental variables related to urbanization. In total, we detected 381 outlier loci and after stringent filtering, identified and annotated 19 candidate loci. Many of the candidate genes were involved in metabolic processes and have well-established roles in metabolizing lipids and carbohydrates. Our results indicate that white-footed mice in New York City are adapting at the biomolecular level to local selective pressures in urban habitats. Annotation of outlier loci suggests selection is acting on metabolic pathways in urban populations, likely related to novel diets in cities that differ from diets in less disturbed areas.

Effects of parasite and historic driven selection on the diversity and structure of a MHC-II gene in a small mammal species (Peromyscus leucopus) undergoing range expansion

Genetic diversity may decrease from the centre to the margin of a species distribution range due to neutral stochastic processes. Selection may also alter genetic diversity in non-neutral markers, such as genes associated with the immune system. Both neutral processes and selection on the immune system are thus expected to affect the spatial distribution of such markers, but the relative strength of each has been scarcely studied. Here, we compared the diversity of a neutral marker (mitochondrial cytochrome b) and a selected marker (DRB gene from the MHC-II), in eastern-North American populations of white-footed mice (Peromyscus leucopus), a species known for its role of main reservoir of the Lyme disease. We observed distinct phylogeographic patterns with these two markers, which may be the result of selection pressure acting upon the DRB gene. As predicted by the central marginal hypothesis, we observed a loss of neutral genetic diversity toward the margin of the species distribution. A decrease in diversity was also observed for the DRB gene, likely due to genetic drift and positive selection operated by helminth parasites. Such a loss in genetic diversity at the range margin may slow down the ongoing expansion of P. leucopus, by counterbalancing the effect of global warming on the mouse survival at higher latitude.

Inbreeding and selection shape genomic diversity in captive populations: Implications for the conservation of endangered species

Captive breeding programs are often initiated to prevent species extinction until reintroduction into the wild can occur. However, the evolution of captive populations via inbreeding, drift, and selection can impair fitness, compromising reintroduction programs. To better understand the evolutionary response of species bred in captivity, we used nearly 5500 single nucleotide polymorphisms (SNPs) in populations of white-footed mice (Peromyscus leucopus) to measure the impact of breeding regimes on genomic diversity. We bred mice in captivity for 20 generations using two replicates of three protocols: random mating (RAN), selection for docile behaviors (DOC), and minimizing mean kinship (MK). The MK protocol most effectively retained genomic diversity and reduced the effects of selection. Additionally, genomic diversity was significantly related to fitness, as assessed with pedigrees and SNPs supported with genomic sequence data. Because captive-born individuals are often less fit in wild settings compared to wild-born individuals, captive-estimated fitness correlations likely underestimate the effects in wild populations. Therefore, minimizing inbreeding and selection in captive populations is critical to increasing the probability of releasing fit individuals into the wild.

Genetic variation in total number and locations of GnRH neurons identified using in situ hybridization in a wild-source population.

The evolution of brain function in the regulation of physiology may depend in part upon the numbers and locations of neurons. Wild populations of rodents contain natural genetic variation in the inhibition of reproduction by winter-like short photoperiod, and it has been hypothesized that this functional variation might be due in part to heritable variation in the numbers or location of gonadotropin releasing hormone (GnRH) neurons. A naturally variable wild-source population of white-footed mice was used to develop lines artificially selected for or against mature gonads in short, winter-like photoperiods. We compared a selection line that is reproductively inhibited in short photoperiod (Responsive) to a line that is weakly inhibited by short photoperiod (Nonresponsive) for differences in counts of neurons identified using in situ hybridization for GnRH mRNA. There was no effect of photoperiod, but there were 60% more GnRH neurons in total in the Nonresponsive selection line than the Responsive selection line. The lines differed specifically in numbers of GnRH neurons in more anterior regions, whereas numbers of GnRH neurons in posterior areas were not statistically different between lines. We compare these results to those of an earlier study that used immunohistochemical labeling for GnRH neurons. The results are consistent with the hypothesis that the selection lines and natural source population contain significant genetic variation in the number and location of GnRH neurons. The variation in GnRH neurons may contribute to functional variation in fertility that occurs in short photoperiods in the laboratory and in the wild source population in winter.

Genetic variation in male sexual behaviour in a population of white-footed mice in relation to photoperiod

In natural populations, genetic variation in seasonal male sexual behaviour could affect behavioural ecology and evolution. In a wild-source population of white-footed mice, Peromyscus leucopus, from Virginia, U.S.A., males experiencing short photoperiod show high levels of genetic variation in reproductive organ mass and neuroendocrine traits related to fertility. We tested whether males from two divergent selection lines, one that strongly suppresses fertility under short photoperiod (responder) and one that weakly suppresses fertility under short photoperiod (nonresponder), also differ in photoperiod-dependent sexual behaviour and responses to female olfactory cues. Under short, but not long, photoperiod, there were significant differences between responder and nonresponder males in sexual behaviour and likelihood of inseminating a female. Males that were severely oligospermic or azoospermic under short photoperiod failed to display sexual behaviour in response to an ovariectomized and hormonally primed receptive female. However, on the day following testing, females were positive for spermatozoa only when paired with a male having a sperm count in the normal range for males under long photoperiod. Males from the nonresponder line showed accelerated reproductive development under short photoperiod in response to urine-soiled bedding from females, but males from the responder line did not. The results indicate genetic variation in sexual behaviour that is expressed under short, but not long, photoperiod, and indicate a potential link between heritable neuroendocrine variation and male sexual behaviour. In winter in a natural population, this heritable behavioural variation could affect fitness, seasonal life history trade-offs and population growth.

The lyme disease pathogen has no effect on the survival of its rodent reservoir host.

Zoonotic pathogens that cause devastating morbidity and mortality in humans may be relatively harmless in their natural reservoir hosts. The tick-borne bacterium Borrelia burgdorferi causes Lyme disease in humans but few studies have investigated whether this pathogen reduces the fitness of its reservoir hosts under natural conditions. We analyzed four years of capture-mark-recapture (CMR) data on a population of white-footed mice, Peromyscus leucopus, to test whether B. burgdorferi and its tick vector affect the survival of this important reservoir host. We used a multi-state CMR approach to model mouse survival and mouse infection rates as a function of a variety of ecologically relevant explanatory factors. We found no effect of B. burgdorferi infection or tick burden on the survival of P. leucopus. Our estimates of the probability of infection varied by an order of magnitude (0.051 to 0.535) and were consistent with our understanding of Lyme disease in the Northeastern United States. B. burgdorferi establishes a chronic avirulent infection in their rodent reservoir hosts because this pathogen depends on rodent mobility to achieve transmission to its sedentary tick vector. The estimates of B. burgdorferi infection risk will facilitate future theoretical studies on the epidemiology of Lyme disease.

The impacts of inbreeding, drift and selection on genetic diversity in captive breeding populations.

The goal of captive breeding programmes is often to maintain genetic diversity until re-introductions can occur. However, due in part to changes that occur in captive populations, approximately one-third of re-introductions fail. We evaluated genetic changes in captive populations using microsatellites and mtDNA. We analysed six populations of white-footed mice that were propagated for 20 generations using two replicates of three protocols: random mating (RAN), minimizing mean kinship (MK) and selection for docility (DOC). We found that MK resulted in the slowest loss of microsatellite genetic diversity compared to RAN and DOC. However, the loss of mtDNA haplotypes was not consistent among replicate lines. We compared our empirical data to simulated data and found no evidence of selection. Our results suggest that although the effects of drift may not be fully mitigated, MK reduces the loss of alleles due to inbreeding more effectively than random mating or docility selection. Therefore, MK should be preferred for captive breeding. Furthermore, our simulations show that incorporating microsatellite data into the MK framework reduced the magnitude of drift, which may have applications in long-term or extremely genetically depauperate captive populations.

Patterns of Mortality in a Wild Population of White-Footed Mice

Abstract Understanding causes of mortality in a population provides information about the selection pressures acting on individuals in that population. To obtain unbiased estimates of mortality, researchers must track individual animals until they die, retrieve dead animals, and make a forensic estimation to determine the cause. Given the limitations of traditional radio-tracking technology and the biases inherent in other methods of estimating mortality causes, little is known about causes of mortality in small mammals (<1 kg). Here we used an automated radio telemetry system to record activity patterns and detect mortality events in a wild population of Peromyscus leucopus (White-footed Mouse). Mice (n = 32) were fitted with small (1 g) radio-collars. Daily survival rate was 0.98 ± 0.01, which translates to a 50% chance of survival over a 29-day period. Although the species is well known as a vector for various diseases, we found only one death attributable to disease. Most mouse mortality (93%) was cau...

Heritable variation in reaction norms of metabolism and activity across temperatures in a wild-derived population of white-footed mice (Peromyscus leucopus)

Heritable variation in metabolic traits is likely to affect fitness. In this study, white-footed mice from wild-derived photoresponsive [R, infertile in short day length (SD)] and non-photoresponsive (NR, fertile in SD) selection lines were maintained under short-day (SD 8Light:16Dark), sub-thermoneutral conditions (22 or 12°C). Mice had significantly higher levels of food intake and resting metabolic rates (RMR) at low temperature. RMR differed significantly between lines (greater in NR mice). In contrast to previous work under thermoneutral conditions, there was no significant difference in overall activity or average daily metabolic rates (ADMR) of mice from the two lines. Reduced activity may reflect behavioral changes under cooler conditions (e.g., nest building) reducing the overall energetic cost of fertility (for NR mice). There was no significant difference in maximal rate of oxygen consumption ([Formula: see text]) between lines. R mice had significantly greater brown adipose tissue and white abdominal fat mass due to both line and temperature. Reaction norms for intake, resting metabolism (RMR/BMR) and level of activity from current (12 and 22°C) and previously published data (28°C) demonstrate independent effects of genetics (line) and environment (temperature) for resting metabolism, but a clear interaction between these for activity. The results suggest that fertility under winter conditions imposes metabolic costs that are related to the level of reproductive development. Under the coldest conditions tested, however, mice that remained fertile in SD reduced activity, ADMR and food requirements, decreasing the differential between selection lines. Heritable variation in reaction norms suggests a genetic by environment effect that could be subject to selection.

Congruent morphological and genetic differentiation as a signature of range expansion in a fragmented landscape.

Phenotypic differentiation is often interpreted as a result of local adaptation of individuals to their environment. Here, we investigated the skull morphological differentiation in 11 populations of the white-footed mouse (Peromyscus leucopus). These populations were sampled in an agricultural landscape in the Montérégie region (Québec, Canada), at the northern edge of the distribution of the white-footed mouse. We found a strong pattern of phenotypic differentiation matching the genetic structure across these populations. Landscape fragmentation and the presence of geographic barriers, in particular north–south oriented rivers, contribute to this differentiation and modulate the pattern of rapid ongoing northward range expansion of the white-footed mouse in response to climate warming. We conclude that while large rivers and postglacial recolonization routes have shaped the current pattern of distribution and differentiation of white-footed mouse populations, further local differentiation is occurring, at the scale of the landscape. We posit that the northern expansion of the white-footed mouse is achieved through successive independent founder events in a fragmented landscape at the northern range edge of the species. The phenotypic differentiation we observe is thus a result of a number of mechanisms operating at different spatial and temporal scales.

Demographic Effects of White-tailed Deer (Odocoileus virginianus) Exclosures on White-footed Mice (Peromyscus leucopus)

In 1994 two 0.65 ha deer exclosures were built in a second growth forest in northeastern Pennsylvania, one under an oak-maple canopy and the other under eastern hemlock. We hypothesized that populations of white-footed mice (Peromyscus leucopus) outside the exclosures were depressed because of herbaceous cover removal by white-tailed deer (Odocoileus virginianus) and/or by acorn competition with deer and would be higher inside the exclosures. For 10 y, the small mammal communities were censured with one 10 × 4 Sherman live-trap grid in each exclosure and one control grid of identical dimensions immediately outside each exclosure. Paired-T comparisons between the oak-maple exclosure and control grids, and the hemlock exclosure and control grids indicated that more total males were captured in the oak-maple exclosure grid than in the oak-maple control grid. Although repeated-measures ANOVA indicated that habitat (tree canopy type) produced more differences in mice numbers than did the presence of the exclosures, it also demonstrated that numbers of large females fluctuated less in the oak-maple exclosure than in the corresponding control. A repeated measures ANOVA of Jolly-Seber population estimate indicated that more mice were trapped in the combined oak-maple and hemlock exclosure grids than in the combined control grids.

Impacts of generalist mesopredators on the demography of small-mammal populations in fragmented landscapes

A consequence of the reduction and subsequent fragmentation of native habitats has been the loss or severe reduction of specialist predator populations from these altered ecosystems, resulting in a “release” of generalist predators. Demographic aspects of small-rodent populations, especially predator-driven density cycles, have been extensively studied. However, the majority of studies examining predator–prey dynamics have been conducted in relatively undisturbed ecosystems, while more limited data are available for regions that have been greatly modified by human settlement. Using raccoons ( Procyon lotor (L., 1758)) and white-footed mice ( Peromyscus leucopus (Rafinesque, 1818)) as focal species, we used an experimental framework to evaluate the hypothesis that generalist mesopredators limit small-mammal abundance in landscapes that have been significantly altered by human land use. Both parametric and nonparametric analyses indicated that populations of white-footed mice exhibited a significant increase (32%) in density where raccoon abundance was reduced when compared with control populations. Our study highlights an important role that superabundant mesopredators can play in ecosystems through the limitation of secondary prey populations. This research suggests that further investigation of the trophic dynamics of agricultural ecosystems is critical if we are to elucidate the fundamental ecological mechanisms associated with the persistence of species in disturbed environments.

Tissue–diet discrimination factors and turnover of stable carbon and nitrogen isotopes in white-footed mice (Peromyscus leucopus)

Stable isotope analysis has become an increasingly valuable tool in investigating animal ecology. Here we document the turnover rates for carbon in the liver, muscle, and whole blood tissue, as well as the tissue–diet discrimination values for carbon and nitrogen isotopes in the liver, whole blood, muscle, and hair, of the white-footed mouse ( Peromyscus leucopus (Rafinesque, 1818)). A 168-day diet-switching experiment was conducted with a laboratory population of white-footed mice. The δ13C values for all tissues deviated less than 1‰ from those of the diet except for whole blood, which had a slightly higher tissue–diet discrimination factor of 1.8‰. All tissues were enriched in 15N by approximately 3‰ relative to the diet except for liver tissue, which was 4.5‰ higher than the dietary δ15N value. Turnover rates for tissues of white-footed mice were ranked liver &gt; whole blood &gt; muscle. The half-lives calculated for liver tissue differed significantly between the two diet switches performed in this experiment. We demonstrate that there is potential for variation in tissue–diet discrimination values and tissue turnover rates between even closely related species. These findings highlight the importance of determining species-specific estimates of these parameters prior to the use of stable isotope analysis in field investigations of animal ecology.

Rapid, pervasive genetic differentiation of urban white-footed mouse (Peromyscus leucopus) populations in New York City

We investigated genetic diversity and structure of urban white-footed mouse, Peromyscus leucopus, populations in New York City (NYC) using variation at 18 microsatellite loci. White-footed mice are 'urban adapters' that occur at higher population densities as habitat fragments are reduced in area but have a limited ability to disperse through urbanized areas. We hypothesized that this combination of traits has produced substantial genetic structure but minimal loss of genetic variation over the last century in NYC. Allelic diversity and heterozygosity in 14 NYC populations were high, and nearly all of our NYC study sites contained genetically distinct populations of white-footed mice as measured by pairwise FST , assignment tests, and Bayesian clustering analyses performed by Structure and baps. Analysis of molecular variance revealed that genetic differences between populations separated by a few kilometres are more significant than differences between prehistorically isolated landmasses (i.e. Bronx, Queens, and Manhattan). Allele size permutation tests and lack of isolation by distance indicated that mutation and migration are less important than drift as explanations for structure in urban, fragmented P.leucopus populations. Peromyscus often exhibit little genetic structure over even regional scales, prompting us to conclude that urbanization is a particularly potent driver of genetic differentiation compared to natural fragmentation.

Variation in levels of luteinizing hormone and reproductive photoresponsiveness in a population of white-footed mice (Peromyscus leucopus)

Natural genetic variation in reproduction and life history strategies is a manifestation of variation in underlying regulatory neuronal and endocrine systems. A test of the hypothesis that genetic variation in luteinizing hormone (LH) level could be related to a life history trait, seasonal reproduction, was conducted on artificial selection lines from a wild-source population of white-footed mice (Peromyscus leucopus). Variation exists in the degree of suppression of reproduction by winter short-day photoperiods (SD) in wild-source individuals and in the laboratory population. In this population, most individuals from a photoperiod-responsive (R) artificial selection line are strongly suppressed reproductively in SD, while most individuals from a photoperiod-nonresponsive (NR) artificial selection line are only weakly reproductively suppressed in SD. We assayed levels of LH to test for genetic variation between lines that could contribute to variation in reproductive status in SD. Females from both lines were raised in long-day photoperiods (LD) or SD, ovariectomized under isoflurane anesthesia, and given estradiol implants. Levels of LH were significantly higher in the NR line than in the R line, indicating genetic variation for levels of LH. Levels of LH were higher in LD than in SD, indicating that levels of LH were sensitive to photoperiod treatment even with a controlled level of estradiol negative feedback. The results indicate that there is genetic variation in levels of LH that could be functionally important both in the laboratory in SD and in the wild population in winter. Thus genetic variation in levels of LH is a plausible causal factor determining winter reproductive phenotype in the wild population.

Microevolution of neuroendocrine mechanisms regulating reproductive timing in Peromyscus leucopus

A key question in the evolution of life history and in evolutionary physiology asks how reproductive and other life-history traits evolve. Genetic variation in reproductive control systems may exist in many elements of the complex inputs that can affect the hypothalamic-pituitary-gonadal (HPG) or reproductive axis. Such variation could include numbers and other traits of secretory cells, the amount and pattern of chemical message released, transport and clearance mechanisms, and the number and other traits of receptor cells. Selection lines created from a natural population of white-footed mice (Peromyscus leucopus) that contains substantial genetic variation in reproductive inhibition in response to short winter daylength (SD) have been used to examine neuroendocrine variation in reproductive timing. We hypothesized that natural genetic variation would be most likely to occur in the inputs to GnRH neurons and/or in GnRH neurons themselves, but not in elements of the photoperiodic pathway that would have pleiotropic effects on nonreproductive functions as well as on reproductive functions. Significant genetic variation has been found in the GnRH neuronal system. The number of GnRH neurons immunoreactive to an antibody to mature GnRH peptide under conditions maximizing detection of stained neurons was significantly heritable in an unselected control (C) line. Furthermore, a selection line that suppresses reproduction in SD (photoperiod responsive, R) had fewer IR-GnRH neurons than a selection line that maintains reproduction in SD (photoperiod nonresponsive, NR). This supports the hypothesis that genetic variation in characteristics of GnRH neurons themselves may be responsible for the observed phenotypic variation in reproduction in SD. The R and NR lines differ genetically in food intake and iodo-melatonin receptor binding, as well as in other characteristics. The latter findings are consistent with the hypothesis that genetic variation occurs in the nutritional and hormonal inputs to GnRH neurons. Genetic variation also exists in the phenotypic plasticity of responses to two combinations of treatments, (1) food and photoperiod, and (2) photoperiod and age, indicating genetic variation in individual norms of reaction within this population. Overall, the apparent multiple sources of genetic variation within this population suggest that there may be multiple alternative combinations of alleles for both the R and NR phenotypes. If that interpretation is correct, we suggest that this offers some support for the evolutionary "potential" hypothesis and is inconsistent with the evolutionary "constraint" and "symmorphosis" hypotheses for the evolution of complex neuroendocrine pathways.