Mus Spretus Research Articles

Integrated application of transcriptomics, proteomics, and metallomics in environmental studies

Abstract Here we report a preliminary working scheme for the integrative application of transcriptomic, proteomic, and metallomic methodologies in environmental monitoring, by using as sentinel the wildlife species Mus spretus and as reference the gene/protein sequence databases from the key model species Mus musculus. We have demonstrated that the absolute transcript expression signatures quantified by reverse transcription (RT) and real-time polymerase chain reaction (PCR) of selected key genes (e.g., those coding for biotransformation enzymes) in M. spretus is a useful and reliable novel biomonitoring end-point. The suitability of commercial M. musculus oligonucleotide arrays for genome-wide transcriptional profiling in M. spretus has been also shown. Transcriptomic studies indicate considerable gene sequence similarities between both mouse species. Based on these similarities, we have demonstrated the applicability in free-living M. spretus of high-throughput proteomic methods, based on matrix-assisted laser desorption/ionization with time-of-flight mass spectrometry (MALDI-TOFMS) analysis of tryptic 2D electrophoresis (2-DE) spot digest and peptide matching with M. musculus database. A metallomic approach based on size exclusion chromatography inductively coupled plasma-mass spectrometry (SEC-ICP-MS) was applied to trace metal-biomolecule profiles. A preliminary integration of these three -omics has been addressed to M. musculus/M. spretus couple, two rodent species that separated 3 million years ago. The integrated application of transcriptomic and proteomic data and the bidirectional use of metallomics and proteomics for selective isolation of metal-biomolecules are covered in the working scheme MEPROTRANS-triple-OMIC reported in this study.

Gene expression regulation in the context of mouse interspecific mosaic genomes

The testis transcriptome of mouse strains containing homozygous segments of Mus spretus origin in a Mus musculus background was analyzed.

Interspecific Recombinant Congenic Strains Between C57BL/6 and Mice of the Mus spretus Species: A Powerful Tool to Dissect Genetic Control of Complex Traits

Complex traits are under the genetic control of multiple genes, often with weak effects and strong epistatic interactions. We developed two new collections of mouse strains to improve genetic dissection of complex traits. They are derived from several backcrosses of the Mus spretus SEG/Pas or STF/Pas strains on the C57BL/6J background. Each of the 55 interspecific recombinant congenic strains (IRCSs) carries up to eight SEG/Pas chromosomal segments with an average size of 11.7 Mb, totalizing 1.37% of the genome. The complete series covers 39.7% of the SEG/Pas genome. As a complementary resource, six partial or complete interspecific consomic strains were developed and increased genome coverage to 45.6%. To evaluate the usefulness of these strains for QTL mapping, 16 IRCSs were compared with C57BL/6J for seven hematological parameters. Strain 66H, which carries three SEG/Pas chromosomal segments, had lower red blood cell volume and higher platelet count than C57BL/6J. Each chromosomal segment was isolated in a congenic strain to evaluate individual effects. Congenic strains were combined to assess epistasis. Our data show that both traits were controlled by several genes with complex epistatic interactions. IRCSs are therefore useful to unravel QTL with small effects and gene-by-gene interactions.

Proteomics in free‐living Mus spretus to monitor terrestrial ecosystems

We evaluated the suitability of high-throughput proteomic methods to monitor terrestrial ecosystems. Free-living Mus spretus from three sites along the "Domingo Rubio" (DR) stream were compared with mice from Doñana Biological Reserve ("Santa Olalla" lagoon (SOL) negative control), using specimens from an industrial settlement (phosphogypsum stacks (PS)) and rice fields ("Matochal" rice fields (ARZ)) as positive controls. Our 2-DE analysis showed 36 spots with significantly altered expression. Sixteen were identified by MALDI-TOF-PMF and peptide matching with Mus musculus databases. Identified proteins play different roles: cytoskeletal dynamics, proteolysis, biotransformation, oxidative-stress adaptation, and metabolism. Animals from different polluted environments showed contrasting differences in their proteomes, with specific increases and decreases in selected groups of proteins that seem to be co-ordinately regulated. Proteomic data were consistent with metal biomonitoring and conventional biomarker responses, indicating that DR (and PS/ARZ) animals sustained a heavier pollutant burden than SOL specimens and suffered a chronic oxidative stress. Whereas some protein expression differences may protect mice from pollutant toxicity, others should make them more susceptible. Transcript expression signatures agree with the documented lack of correlation between mRNA and protein levels. Nonetheless, a positive significant correlation was found between the gpx1 mRNA molecules and the intensity of one of the two identified GPX1 isospots.

Seed choice by rodents: learning or inheritance?

Learning plays a central role in animal life, and it has received special attention in the context of foraging. In this study, we have tested whether learning operates in seed choices by rodents using the Algerian mouse (Mus spretus) and Holm oak (Quercus ilex) acorns as a model. At the laboratory, those rodents captured in the field during the acorn fall period (experienced individuals) rejected weevil-infested acorns, presumably because of their lower energetic value. By contrast, rodents born in captivity and reared without any contact with acorns (naives) predated weevil-infested acorns at similar rates than sound ones. After exposing naives to infested and sound acorns during 15 days, they rejected infested acorns as the experienced individuals. In the field, predation on weevil-infested acorns was lower than on sound ones. Predation rates on infested acorns were lowest at the end of the acorn fall season, whereas predation rates of sound acorns increased along the same period. This might be explained by the improved ability to reject infested acorns because of the accumulated experience acquired by the initially naive rodents along the acorn fall season. We show that learning shapes strongly seed choices by rodents, and it may be advantageous over inherited behaviors in variable unpredictable situations, such as acorn infestation rates that vary strongly between years and trees. We consider that the role of learning has to be taken into account in future studies on seed predation by rodents.

Genotoxicity detected in wild mice living in a highly polluted wetland area in south western Spain

A field study was carried out in the south of the Iberian Peninsula in an industrial area in the neighbourhood of Huelva city, SW Spain, and in a natural area (Doñana National Park) for comparison, to estimate the genetic risk induced by environmental pollution in wild mice. Genotoxic effects in a sentinel organism, the Algerian mice ( Mus spretus) free living in the industrial area were compared with animals of the same species living in the natural protected area. The single cell gel electrophoresis, or Comet assay, was performed as a genotoxicity test in peripheral blood of mice. Our results clearly show that mice free living in the contaminated area bear a high burden of genetic damage as compared with control individuals. The results suggest that the assessing of genotoxicity levels by the Comet assay in wild mice can be used as a valuable test in pollution monitoring and environmental conservation.

Rodents change acorn dispersal behaviour in response to ungulate presence

Small rodents are prominent seed predators, but they also favour plant recruitment as seed dispersers. The direct interactions of ungulates on plants are more one‐sided and negative, as they mainly reduce plant recruitment through predation on seeds and seedlings. The effects of small rodents and ungulates on plant recruitment have been considered and studied as independent episodes within plant regeneration cycles. However, ungulate–rodent interactions and their potential effects on plant regeneration have not been considered so far. A number of studies have recently documented ungulate effects on the abundance, diversity and spatial distribution of small rodents. Here, we hypothesize that ungulates may also affect rodent seed dispersal behaviour. We monitored acorn dispersal by small rodents (Mus spretus and Apodemus sylvaticus) in oak woodlands with and without exclosures for large ungulates, mainly red deer, Cervus elaphus, and wild boar, Sus scrofa. The study was carried out in a typical Mediterranean Holm oak, Quercus ilex, forest throughout the acorn fall season in 2003 and 2004. We found that, in both years, the proportion of acorns cached and not recovered in the short‐term was, on average, lower in the presence (1.4%) than in the absence (19.9%) of ungulates. Acorn dispersal distances were not affected by ungulate presence in either year. However, ungulates had an effect on the spatial distribution of dispersed seeds; rodents apparently avoided shrubs as caching sites in both years. This result was interpreted as a behavioural response to reduce the risk of cache pilferage by conspecifics, which are closely associated with shrubs in presence, but not in absence, of ungulates. Potential effects of different densities of rodents or predators were discarded, as none of them differed between the areas with and without ungulates. The present study found significant interactions between heterospecific seed and seedling consumers that had been considered as independent episodes within tree regeneration cycles. As a result of such interactions, ungulates may have negative indirect effects on oak recruitment by reducing (1) acorn caching frequency, and (2) the proportion of acorns cached under shrubs, key nurse‐plants for the establishment of Holm oak seedlings in Mediterranean areas.

Tint Maps to Mouse Chromosome 6 and May Interact With a Notochordal Enhancer of Brachyury

At the proximal part of mouse chromosome 17 there are three well-defined genes affecting the axis of the embryo and consequently tail length: Brachyury, Brachyury the second, and the t-complex tail interaction (T1, T2, and tct). The existence of T1 and tct in fact defines the classical "t-complex" that occupies approximately 40 cM of mouse chromosome 17. Their relationship to each other and various unlinked interacting genes has been enigmatic. The tint gene was the first of the latter to be identified. We report here its genetic mapping using a microsatellite scan together with outcrosses to Mus spretus and M. castaneous followed by a subsequent testcross to T, T1, and T2 mutants. Surprisingly, tint interacts with T2 but not with T1. The implications of our data suggest that T2 may be part of the T1 regulatory region through direct or indirect participation of tint.

Multigenic control of skin tumour development in mice.

Different inbred mouse strains vary greatly in their susceptibility to tumour development in a variety of tissues. Intraspecific and interspecific crosses can, therefore, be used to map the loci that control this predisposition. Crosses of Mus musculus with Mus spretus are highly resistant to tumour development in the skin, liver, lung and lymphoid system. M. spretus, therefore, has dominantly acting resistance loci, which we have attempted to map. More than 350 interspecific backcross mice were followed for 18 months to assess susceptibility to development of chemically induced papillomas and carcinomas. The results were analysed using a combination of MAPMAKER/QTL analysis and multiple regression analysis for the determination of linkage in multigenic quantitative traits. The results showed clearly that at least three genes on chromosomes 5 and 7 control resistance to tumour development. Importantly, some genes confer resistance to benign tumours but they have relatively little effect on malignant progression. This suggests the existence of different classes of benign tumours: those that are capable of tumour progression and those that have only a very low probability of becoming malignant. Identification of these genes will improve our understanding of mechanisms of carcinogenesis and may provide a novel route to the identification of "low-penetrance' human tumour susceptibility genes.

Pair bonding in the wild mouse Mus spretus: inference on the mating system

The mating system of murids remains largely unknown, with controversial reports. Reevaluation of previous data reveals monogamous features in the Algerian mouse from the South of France, Mus spretus. To test this hypothesis, we performed dyadic encounters toward three kinds of opponents from the other sex (his/her own mate, a mouse from an unknown pair, and an isolated individual), and compared the results to those of a related acknowledgedly polygynous related species, the house mouse M. musculus domesticus. We expected more affiliative behaviours toward its mate from the M. spretus male, and a higher interest toward unknown females in the house mouse. Females should not exhibit striking differences toward their partner, contrary to their behaviour toward unknown males. Results showed the predicted behaviour for males as well as for females: more amicable, but also sexual, toward its mate for M. spretus male, the unknown protagonists eliciting the greatest social interest in the case of the house mouse. To cite this article: J. Cassaing, F. Isaac, C. R. Biologies 330 (2007).

The helminth community of the Mediterranean mouse, Mus spretus, in a post-fire regenerated Mediterranean ecosystem

Abstract The helminth community of the Mediterranean mouse, Mus spretus, was analysed in a post-fire regenerated Mediterranean ecosystem. The study was carried out in the Spanish Natural Park of the Serra Calderona and comprised a 13 year period, from the 2nd to the 14th year after a wildfire. A total of 121 host individuals was analysed, 66 mice from the burned area and 55 from the non-burned area used as control. The results show a helminth community consisting of 10 helminth species, characterised by low diversity, with Syphacia obvelata as the only dominant helminth species. The helminth infracommunity, determined by its origin of capture, burned or non-burned areas, shows some significant differences. It is concluded that the helminth community of the Mediterranean mouse could be considered as a potential biological tag of the post-fire regeneration in Mediterranean ecosystems and therefore might explain some of the changes occurring and their repercussions in perturbed areas.

Eye Morphology in some Wild Rodents

The eye anatomy of six rodent species (Murinae: Apodemus sylvaticus, Mus domesticus, and Mus spretus; Arvicolinae: Clethrionomys glareolus, Arvicola terrestris and Microtus arvalis) was compared by means of light or electron microscopy to determine adaptive, and evolutive signals. Our observations revealed inter-specific morphological differences, which were moderate among representatives of the same subfamily. Specifically, traits that distinguished murines from arvicolines were the globe's relative size, the pupillary constrictor muscle, the amount of retinal epithelium melanin, and the thickness of certain ocular coats. Moreover, adaptations to new habitats and differences in temporal activity among species of the same subfamily determined discords respect to the phylogenetic patterns. This was true of the adaptations to underground conditions seen in A. terrestris, which involved the thickness of the cornea, sclera, and choroids. Likewise, A. sylvaticus had adaptations to its nocturnal lifestyle, as shown by the large overall size of the eye and lens, and by a large, thick cornea.

Centimorgan-Range One-Step Mapping of Fertility Traits Using Interspecific Recombinant Congenic Mice

In mammals, male fertility is a quantitative feature determined by numerous genes. Until now, several wide chromosomal regions involved in fertility have been defined by genetic mapping approaches; unfortunately, the underlying genes are very difficult to identify. Here, 53 interspecific recombinant congenic mouse strains (IRCSs) bearing 1-2% SEG/Pas (Mus spretus) genomic fragments disseminated in a C57Bl/6J (Mus domesticus) background were used to systematically analyze male fertility parameters. One of the most prominent advantages of this model is the possibility of analyzing stable phenotypes in living animals. Here, we demonstrate the possibility in one-step fine mapping for several fertility traits. Focusing on strains harboring a unique spretus fragment, we could unambiguously localize two testis and one prostate weight-regulating QTL (Ltw1, Ltw2, and Lpw1), four QTL controlling the sperm nucleus shape (Sh1, Sh2, Sh3, and Sh4), and one QTL influencing sperm survival (Dss1). In several cases, the spretus DNA fragment was small enough to propose sound candidates. For instance, Spata1, Capza, and Tuba7 are very strong candidates for influencing the shape of the sperm head. Identifying new genes implied in mammalian fertility pathways is a necessary prerequisite for clarifying their molecular grounds and for proposing diagnostic tools for masculine infertilities.

New chemically induced skin tumour susceptibility loci identified in a mouse backcross between FVB and dominant resistant PWK

BackgroundA variety of skin cancer susceptibility among mouse strains has allowed identification of genes responsible for skin cancer development. Fifteen Skts loci for skin tumour susceptibility have been mapped so far by using the two-stage skin carcinogenesis model [induced by 7.12-dimethylbenz(a)anthracene (DMBA)/12-O-tetradecanoylphorbol-13-acetate (TPA)]. A few responsible genes have been identified using wild-derived dominant resistant Mus spretus mice, and one has been confirmed as a low penetrance cancer susceptibility gene in a variety of human cancers.ResultsIn the present study, we found that wild-derived PWK mice developed no tumour by treatment with the two-stage skin carcinogenesis protocol. This phenotype is dominant resistant when crossed with the highly susceptible strain FVB. By analyzing the F1 backcross generation between PWK and FVB, we found empirical evidence of significant linkage at the new loci Skts-fp1 on chromosome 4 and suggestive linkage on chromosomes 1, 3, 11, 12 and 14 for skin tumour susceptibility. Skts-fp1 includes the Skts7 interval, which was previously mapped by a Mus spretus and NIH backcross. We also observed suggestive linkage on chromosomes 1 and 2 in the female population only, while suggestive linkage on chromosomes 14 and 15 only was observed in the male population. A significant genetic interaction was seen between markers of D11Mit339 and D16Mit14.ConclusionAnalysis of this new cross may facilitate the identification of genes responsible for mouse skin cancer susceptibility and may reveal their biological interactions.

Evolution of an invasive rodent on an archipelago as revealed by molar shape analysis: the house mouse in the Canary Islands

AbstractAim The aim of this paper is to identify the patterns in the morphological differentiation in Canary Island mice, based on fossil and modern samples. In order to achieve this, the mouse species present on the archipelago were first compared with a set of continental mice. The differences between the continental and Canary Island samples, and among the Canary Island samples, provide insights into the processes of colonization and the subsequent insular evolution.Location Canary archipelago.Methods An outline analysis based on Fourier transformation was used to quantify shape differences between lower molars. Together with the fossil and modern Canary Island samples, a reference set of genotyped continental populations of the commensal Mus musculus and the wild Mus spretus was used for comparison.Results The morphometric analysis showed that all the mouse specimens from the Canary Islands and Cape Verde belonged to Mus musculus domesticus. Lower molars of extant mice from La Gomera, El Hierro, Gran Canaria, Tenerife, and to a lesser degree from Lanzarote, were similar to those of genotyped M. m. domesticus from the continent, while teeth of extant mice from Fuerteventura were more divergent. Fossil mice from Fuerteventura were very similar to the extant representatives on this island, and similar to the fossil mice on the nearby islands of Lobos and La Graciosa.Main conclusions The mouse present on the Canary archipelago has been identified as the house mouse M. m. domesticus. Based on the shape of the lower molar, the Canary Island mice are divergent from the continental ones, but the degree of divergence varies with the geography of the archipelago. Overall, populations from eastern islands are more divergent from the continental mice than populations from western ones. Fossil populations indicate that this situation was established several centuries ago. Two main factors may have contributed to this pattern: the appearance of different types of environment on the islands since the successful settlement of the mouse, and/or the number of subsequent introductions of continental individuals via shipping.

Fine Mapping and Candidate Gene Analyses of Pulmonary Adenoma Resistance 1, a Major Genetic Determinant of Mouse Lung Adenoma Resistance

Pulmonary adenoma resistance 1 (Par1) is a major genetic determinant of mouse lung adenoma resistance. Although Par1 was previously mapped to mouse chromosome 11 by conventional linkage analyses, its candidate region was broad and undefined. In our present study, we generated Par1 congenic mice using two mouse strains A/J (Par1/-) and Mus spretus (Par1/+). Analyzing these congenic mice enabled us to fine map the Par1 quantitative trait loci (QTL) into a 2.0-cM (2.2 Mb) chromosomal region between genetic marker D11Mit70 and the gene Hoxb9. We then conducted systematic candidate gene screening through nucleotide polymorphism and expression analyses. Genes showing differential lung tissue expression or carrying nonsynonymous single nucleotide polymorphisms were identified and discussed. In particular, we evaluated tumor suppressor gene Tob1 for its Par1 candidacy. Our findings have narrowed the Par1 QTL region and will greatly facilitate the identification of the major genetic determinant of mouse lung adenoma resistance.

Habitat associations of small mammals in farmed landscapes: implications for agri-environmental schemes

Abstract The small mammal community in 21 localities of north-western Spain was evaluated in the light of land use composition. The two geomorphologic categories characterising the study area, the main use of the land (arable/pastoral) and main crop types of each sampling locality were used as potential predictors of the relative abundance of five common small mammal species. The Common vole, Microtus arvalis showed a weak relationship with land uses, probably due to the recent colonisation process this species experienced in the study area. The relative abundance of the Algerian mouse, Mus spretus and the Lusitanian pine vole, Microtus lusitanicus was best explained by models built at the broadest regional scale, the former being more abundant in the eastern area, the latter in the western area. The Greater white-toothed shrew, Crocidura russula showed a positive relationship with grassland coverage, whilst the Wood mouse, Apodemus sylvaticus benefited from increasing proportions of fallow lands within the landscape. These two species are then expected to respond positively to those agri-environmental schemes including the increase of fallows and grassy vegetation within the arable landscape (EU recommendations). However, further efforts are needed to predict, at least qualitatively, the response of other small mammal species to the changing farmed landscape. This is especially true for two endemic species occurring at this area: the Cabrera vole, Microtus cabrerae and the Lusitanian pine vole, and for which this kind of information is almost absent.

LIFE CYCLE AND DESCRIPTION OF A NEW SPECIES OF BRACHYLAIMID (TREMATODA: DIGENEA) IN SPAIN

The life cycle of Brachylaima llobregatensis n. sp. (Trematoda: Brachylaimidae) is elucidated. Embryonated, operculated, asymmetric eggs (30.9 x 18.2 microm) are eliminated with feces of natural hosts wood mouse Mus spretus; white-toothed shrew, Crocidura russula; and an experimental host, domestic mouse, Mus musculus var. domesticus. The eggs are ingested by the helicid gastropod Helix (Cornu) aspersa, the only natural and experimental first intermediate host. The miracidium hatches from the egg, infects the snails, and develops into a branched sporocyst in the digestive gland. Microcaudate cercariae emerge from this snail and develop into unencysted metacercariae in the kidney of second intermediate host snails H. (C.) aspersa and Otala punctata (natural hosts) and Theba pisana (experimental host). Ingestion of infected snails leads to the infection of definitive hosts, with the adults inhabiting the middle part of the small intestine. There is a chaetotaxic pattern specific on the acetabular (S(II) 5-6 papillae) and body (papillae absent on P(II)) levels. Three types of cercaria papillae were observed by scanning electron microscopy: argentophilic papillae with fingerlike processes (cephalic, body, and acetabular levels); argentophilic papillae with opening (2 papillae in the M body level); and nonargentophilic dome-shaped papillae (on the cephalic C(II) level, alternating with argentophilic S(II) papillae on the ventral sucker). Suckers are subequal, with the acetabulum located in the posterior part of the anterior third of body. Vitellaria extend from anterior margin of acetabulum to between middle level and anterior margin of anterior testis.

Contrasting evolution of expression differences in the testis between species and subspecies of the house mouse

Regulatory changes in genes involved in reproduction are thought to be prime targets for divergence during speciation, since they are expected to play an important role in sexual selection and sexual conflict. We used microarray analysis of RNA from different wild populations of house mouse subspecies (including Mus m. musculus, Mus m. domesticus, and Mus m. castaneus) and from the sister species Mus spretus to test this assumption. A comparison of expression divergence in brain, liver/kidney, and testis shows a major difference in the evolutionary dynamics of testis-related genes. While the comparison between species confirms an excess in divergence in testis genes, we find that all comparisons between subspecies yield only a very small number of genes with significantly different expression levels in the testis. These results suggest that the early phase of the speciation process may not be driven by regulatory changes in genes that are potential targets of sexual selection, and that the divergence in these genes is only established during a later phase of the speciation process.

Complex genetic control of susceptibility to Mycobacterium bovis (Bacille Calmette-Guérin) infection in wild-derived Mus spretus mice

Susceptibility to Mycobacterium bovis Bacille Calmette-Guérin (BCG) is genetically controlled by Nramp1 (Slc11a1). Inbred mouse strains harbor either the resistance (Nramp1(G169)) or the susceptibility (Nramp1(D169)) allele at Nramp1. Mus spretus (Nramp1(G169); SPRET/EiJ) is shown to display an intermediate level of BCG replication in the spleen (log(10) colony-forming units (CFU) approximately 5), compared to resistant A/J (log(10)CFU approximately 4.0) and susceptible C57BL/6J (log(10)CFU approximately 6.0) mice. The presence of genetic modifiers of Nramp1-dependent susceptibility to M. bovis (BCG) infection in Mus spretus was analyzed by whole-genome scanning in 175 mice of an informative (C57BL/6J x SPRET/EiJ) x C57BL/6J backcross. Nramp1 showed a major effect (D1Mcg4, P<1e(-4)), but additional single marker effects were identified on chromosomes 4 (D4Mit150) and x (DXMit249) in male mice, and on chromosome 9 (D9Mit77) and 17 (D17Mit81) in female mice. A strong interaction between Nramp1 and the major histocompatibility locus was also noted in female mice. The mapped loci may act as modifiers of Nramp1 action, and constitute novel entry points for the parallel search of loci regulating susceptibility to mycobacterial infections in humans.