Reproductive isolation between chromosomal races of the house mouse Mus musculus domesticus in a parapatric contact area revealed by an analysis of multiple unlinked loci

The house mouse, Mus musculus domesticus, exhibits a high level of chromosomal polymorphism because of the occurrence and fast fixation of Robertsonian fusions between telocentric chromosomes. For this reason, it has been considered a classical speciation model to analyse the role of the chromosomal changes in reproductive isolation. In this study, we analysed a parapatric contact area between two metacentric races in central Italy, the Cittaducale race (CD: 2n = 22) and the Ancarano race (ACR: 2n = 24), to estimate gene flow at the boundary. Hybrids between these two races show high levels of structural heterozygosity and are expected to be highly infertile. A sample of 88 mice from 14 sites was used. The mice were genotyped by means of eight microsatellite loci mapped in four different autosomal arms. The results show clear genetic differentiation between the CD and ACR races, as revealed by differences in allele frequencies, factorial correspondence analysis and indexes of genetic population (e.g. F(ST) and R(ST)) along the contact zone. The genetic differentiation between the races was further highlighted by assignation and clustering analyses, in which all the individuals were correctly assigned by their genotypes to the source chromosomal race. This result is particularly interesting in view of the absence of any geographical or ecological barrier in the parapatric contact zone, which occurs within a village. In these conditions, the observed genetic separation suggests an absence of gene flow between the races. The CD-ACR contact area is a rare example of a final stage of speciation between chromosomal races of rodents because of their chromosomal incompatibility.     

The structure of a chromosomal hybrid zone of house mice (Mus domesticus) in central Italy: cytogenetic analysis

Parapatric hybridization between the chromosomal race “CD” (2n = 22) and standard karyotype populations (2n = 40) of Mus domesticus occurs extensively in central Italy. The present paper reports the results of a ctogenetic surve on a transect crossin the hybrid zone north of Rome. No clinal variation in eitier diloid nuders and chromosome frequencies was found to occur in this area, and drift seems to be responsible for the observed atchy pattern of variation. The previous assumption of a strong fertility reduction in structuray heterozyous hybrids contrasts sharply with the width (32 km) of the zone and the average structural aeterozygosity of the hybrid poulations. It is suggested that fitness of structural heterozygotes in nature is not strongly aPfected as has been inferred from laboratory experiments. The results of this study are discussed in context together with the role of hybrid zones in chromosomal speciation in Mus domesticus.     

Phylogenetic analysis of the distribution of chromosomal races of Mus mnscuius domesticus Rutty in Europe

Robertsonian (Rb) translocation is a common chromosomal rearrangement in the house mouse. In free-living populations, 79 fusions with different combinations of chromosomes 1 to 18 have been found in some 45 populations. An updated list of these fusions is presented and analysed in order to reveal the possible processes by which the fusions spread within or among populations. A widespread hypothesis is that when two populations share the same fusion, it can be assumed that they have a common ancestor. This can serve as the basis for the use of the cladistic methods. While I present such an analysis on the updated list of Rbs, I also point to the problems associated with it in this case because many fusions have multiple origins and exchanges of Rbs between populations are frequent. I have tried to use a different approach, based on a critical and quantitative evaluation of the hypothesis of common ancestry. Assuming that the 153 possible fusions have an equal probability of occurrence, I give the formula to compute the probability that populations share a given number of fusions by chance alone. Only when this probability is lower than a chosen level (say 5%) can the populations be inferred to have a non-independent origin (i.e. they have a common ancestor or they have exchanged chromosomes by introgression). This probability measure is then used as a distance estimate to show the relationship between all the Rb populations. This analysis suggests that although some Rbs must have occurred more than once, most of the populations have non-independent origins. Almost all the populations from northern Africa to Belgium and Germany appear to have close karyotopic relationships and form a major group. Clearly independent Rb populations are mainly found in the periphery of this major group, for example in Scotland, Denmark and Spain. 'Chromosomal' flow between Rb populations appears to be a very important process.     

Variation in aggressiveness in house mouse populations

Clearly the ability of 'house mice' to vary their social structures is an important feature contributing to their success in a wide range of habitats. Social structure is strongly influenced by aggressiveness and other behaviours in male and female mice. Material is presented illustrating how genotype, intrauterine location and social experiences influence dyadic encounters in this 'species'.     

Isolation and gene flow: inferring the speciation history of European house mice

Inferring the history of isolation and gene flow during species differentiation can inform us on the processes underlying their formation. Following their recent expansion in Europe, two subspecies of the house mouse (Mus musculus domesticus and Mus musculus musculus) have formed a hybrid zone maintained by hybrid incompatibilities and possibly behavioural reinforcement, offering a good model of incipient speciation. We reconstruct the history of their divergence using an approximate Bayesian computation framework and sequence variation at 57 autosomal loci. We find support for a long isolation period preceding the advent of gene flow around 200,000 generations ago, much before the formation of the European hybrid zone a few thousand years ago. The duration of the allopatric episode appears long enough (74% of divergence time) to explain the accumulation of many post-zygotic incompatibilities expressed in the present hybrid zone. The ancient contact inferred could have played a role in mating behaviour divergence and laid the ground for further reinforcement. We suggest that both subspecies originally colonized the Middle East from the northern Indian subcontinent, domesticus settling on the shores of the Persian Gulf and musculus on those of the Caspian Sea. Range expansions during interglacials would have induced secondary contacts, presumably in Iran, where they must have also interacted with Mus musculus castaneus. Future studies should incorporate this possibility, and we point to Iran and its surroundings as a hot spot for house mouse diversity and speciation studies.     

Gene flow and hybridization following introduction of Mus domesticus into an established population

Seventy-seven house mice ( Mus domesticus ) from Eday, Orkney were released into a long-established population on the Isle of May (56^o 12'N) in 1982. Introduced allozymes, mt and Y-chromosome DNA, and Robertsonian chromosomes spread rapidly, reaching approximate stability c . 3 years later at frequencies different to those in both parental populations. The hybrid population was morphometrically intermediate between the two parents. This is a preliminary summary only; full details will be published elsewhere.     

Bufadienolides in different chromosomal races of Indian squill

Scilliphaeoside and anhydroscilliphaeosidin have been isolated from tetraploid Indian squill. The absence of scilliphaeoside in all populations of diploids and hexaploids and anhydrophaeosidin in diploids, triploids and hexaploids might be one of the reasons why they were not detected in previous studies.     

Re-examination of a proposed case of stasipatric speciation: phylogeography of the Australian morabine grasshoppers (Vandiemenella viatica species group)

Karyotypic differences have been used for delimiting populations or species, although whether these mutations provide strong barriers to gene flow between populations and promote speciation remains contentious. In this study, we assessed whether 11 chromosomal races of Australian morabine grasshoppers ( Vandiemenella viatica species group) represent genetically distinct populations by analyses of cytological and allozyme (35 loci) data and DNA sequences of the elongation factor-1 alpha ( EF -1α), anonymous Mvia11 , and mitochondrial cytochrome c oxidase subunit I ( COI ) loci. While the Vandiemenella chromosomal taxa generally represent genetically distinct units, a substantial portion of the total genetic variation in our samples was not explained by the chromosomal variation. Mantel tests indicated that Vandiemenella populations were spatially structured and have maintained gene flow at a local scale within each of the taxa. The group was subdivided into 13 genetic clusters; four chromosomal taxa comprised single exclusive clusters, while others comprised more than one cluster or clusters shared with other taxa. Boundaries of these cryptic population subdivisions correspond with several biogeographical barriers, such as straits, gulfs, the Murray River, and an ancient mega-lake, Lake Bungunnia. The viatica species group was previously proposed to have diversified without major geographical separation based on the stasipatric speciation model; however, the present study suggests the involvement of allopatric fragmentation. Given extensive nonmonophyly of chromosomal taxa and incomplete barriers to gene flow among taxa, all Vandiemenella chromosomal taxa and genetically distinct populations within chromosomal taxa, except Vandiemenella pichirichi , should be regarded as populations of one species: Vandiemenella viatica .     

Spatio-temporal variation in the structure of a chromosomal polymorphism zone in the house mouse

Several long-term temporal analyses of the structure of Robertsonian (Rb) hybrid zones in the western house mouse, Mus musculus domesticus, have been performed. Nevertheless, the detection of gradual or very rapid variations in a zone may be overlooked when the time elapsed between periods of study is too long. The Barcelona chromosomal polymorphism zone of the house mouse covers about 5000, km(2) around the city of Barcelona and is surrounded by 40 chromosome telocentric populations. Seven different metacentrics and mice with diploid numbers between 27 and 40 chromosomes and several fusions in heterozygous state (from one to seven) have been reported. We compare the present (period 2008-2010) and past (period 1996-2000) structure of this zone before examining its dynamics in more detail. Results indicate that there is not a Rb race in this area, which is consistent with the proposal that this zone was probably originated in situ, under a primary intergradation scenario. The lack of individuals with more than five metacentrics in heterozygous state in the current period suggests that selection acted against such mice. By contrast, this situation did not occur for mice with fewer than five fusions in heterozygous condition. Changes in human activity may affect the dynamics of gene flow between subpopulations, thus altering the chromosomal composition of certain sites. Although these local variations may have modified the clinal trend for certain metacentrics, the general staggered structure of the zone has not varied significantly in a decade.     

Asymmetrical reproductive character displacement in the house mouse

Our study addressed reproductive character displacement between two subspecies of the house mouse, Mus musculus musculus and Mus musculus domesticus that hybridize in Europe along a zone where selection against hybridization is known to occur. Based on a multi-population approach, we investigated spatial patterns of divergence of mate preference in the two taxa. Mate preference was significantly higher in the contact zone than in allopatry in both subspecies, suggesting that reproductive character displacement occurs. Moreover, patterns of preference were stronger in M. m. musculus than in M. m. domesticus, indicating an asymmetrical divergence between the two. In the context of selection against hybridization, our results may provide empirical support for the hypothesis of reinforcement in a parapatric hybrid zone. We discuss factors that could explain the asymmetrical pattern of divergence and the possible impact of a unimodal structure on the maintenance of premating divergence between the two subspecies.     

On the origin and colonization of house mice in the Madeira Islands

The skulls and skins of adult house mice from the Madeira Islands have been studied and compared with those from the Salvage Islands and with material from the neighbouring Portuguese mainland man-associated and wild forms, respectively Mus musclus domesticus Rutty, 1772 and M. sprelus Lataste, 1883. Differences between island and mainland populations were found in some of the analysed features. Insular skins of mice were found to be smaller than those of specimens from the mainland. However, in Madeiran and Salvage mice the toothrows were much more developed than in the mainland house mice. It is considered that the causes of these differences lie in the different characteristics of the habitats, mainly food availability, and also in the isolation of populations. Mus musculus domesticus appears to be the only form of the house mouse to have so far successfully colonized the Madeiras.     

Inferences of selection and migration in the Danish house mouse hybrid zone

We analysed the patterns of allele frequency change for ten diagnostic autosomal allozyme loci in the hybrid zone between the house mouse subspecies Mus musculus domesticus and M. m. musculus in central Jutland. After determining the general orientation of the clines of allele frequencies, we analysed the cline shapes along the direction of maximum gradient. Eight of the ten clines are best described by steep central steps with coincident positions and an average width of 8.9 km (support limits 7.6–12.4) flanked by tails of introgression, indicating the existence of a barrier to gene flow and only weak selection on the loci studied. We derived estimates of migration from linkage disequilibrium in the centre of the zone, and by applying isolation by distance methods to microsatellite data from some of these populations. These give concordant estimates of σ =  0.5–0.8 km generation     . The barrier to gene flow is of the order of 20 km (support limits 14–28), and could be explained by selection of a few per cent at 43–120 underdominant loci that reduces the mean fitness in the central populations to 0.45. Some of the clines appear symmetrical, whereas others are strongly asymmetrical, and two loci appear to have escaped the central barrier to gene flow, reflecting the differential action of selection on different parts of the genome. Asymmetry is always in the direction of more introgression into musculus , indicating either a general progression of domesticus into the musculus territory, possibly mediated by differential behaviour, or past movement of the hybrid zone in the opposite direction, impeded by potential geographical barriers to migration in domesticus territory.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 84 , 593–616.     

The genetic mosaic suggests a new role for hitchhiking in ecological speciation

Early in ecological speciation, the genomically localized effects of divergent selection cause heterogeneity among loci in divergence between incipient species. We call this pattern of genomic variability in divergence the 'genetic mosaic of speciation'. Previous studies have used F(ST) outliers as a way to identify divergently selected genomic regions, but the nature of the relationship between outlier loci and quantitative trait loci (QTL) involved in reproductive isolation has not yet been quantified. Here, we show that F(ST) outliers between a pair of incipient species are significantly clustered around QTL for traits that cause ecologically based reproductive isolation. Around these key QTL, extensive 'divergence hitchhiking' occurs because reduced inter-race mating and negative selection decrease the opportunity for recombination between chromosomes bearing different locally adapted QTL alleles. Divergence hitchhiking is likely to greatly increase the opportunity for speciation in populations that are sympatric, regardless of whether initial divergence was sympatric or allopatric. Early in ecological speciation, analyses of population structure, gene flow or phylogeography based on different random or arbitrarily chosen neutral markers should be expected to conflict--only markers in divergently selected genomic regions will reveal the evolutionary history of adaptive divergence and ecologically based reproductive isolation. Species retain mosaic genomes for a very long time, and gene exchange in hybrid zones can vary dramatically among loci. However, in hybridizing species, the genomic regions that affect ecologically based reproductive isolation are difficult to distinguish from regions that have diverged for other reasons.     

Differential gene flow of mitochondrial and nuclear DNA markers among chromosomal races of Australian morabine grasshoppers (Vandiemenella, viatica species group)

Recent theoretical developments have led to a renewed interest in the potential role of chromosomal rearrangements in speciation. Australian morabine grasshoppers (genus Vandiemenella, viatica species group) provide an excellent study system to test this potential role of chromosomal rearrangements because they show extensive chromosomal variation and formed the basis of a classic chromosomal speciation model. There are three chromosomal races, viatica19, viatica17, and P24(XY), on Kangaroo Island, South Australia, forming five parapatric populations with four putative contact zones among them. We investigate the extent to which chromosomal variation among these populations may be associated with barriers to gene flow. Population genetic and phylogeographical analyses using 15 variable allozyme loci and the elongation factor-1alpha (EF-1alpha) gene indicate that the three races represent genetically distinct taxa. In contrast, analyses of the mitochondrial cytochrome c oxidase subunit I (COI) gene show the presence of three distinctive and geographically localized groups that do not correspond with the distribution of the chromosomal races. These discordant population genetic patterns are likely to result from introgressive hybridization between the chromosomal races and range expansions/contractions. Overall, these results suggest that reduction of nuclear gene flow may be associated with chromosomal variation, or underlying genetic variation linked with chromosomal variation, whereas mitochondrial gene flow appears to be independent of this variation in these morabine grasshoppers. The identification of an intact contact zone between P24(XY) and viatica17 offers considerable potential for further investigation of molecular mechanisms that maintain distinct nuclear genomes among the chromosomal races.     

Microsatellite variability in wild populations of the house mouse is not influenced by differences in chromosomal recombination rates

Differences in recombination rates along the chromosomes can influence the evolution of neutral loci via hitchhiking effects. Generally, these effects should be stronger in regions of low recombination than in regions of high recombination. Detailed information on physical and genetic maps in the house mouse now allows an assessment of the correlation between neutral variability and recombination rates at given chromosomal regions. We chose 29 microsatellite loci from chromosomal regions which show differences in recombination rates and tested their variability in samples from five wild populations of Mus musculus musculus and M. m. domesticus . Our results provide no evidence for a correlation between microsatellite variability and recombination rates. This suggests that the high average mutation rate of microsatellites in mammals counterbalances the effects of long-range hitchhiking in the mouse genome.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 84 , 629–635.     

Hybridization, ecological races and the nature of species: empirical evidence for the ease of speciation

Species are generally viewed by evolutionists as 'real' distinct entities in nature, making speciation appear difficult. Charles Darwin had originally promoted a very different uniformitarian view that biological species were continuous with 'varieties' below the level of species and became distinguishable from them only when divergent natural selection led to gaps in the distribution of morphology. This Darwinian view on species came under immediate attack, and the consensus among evolutionary biologists today appears to side more with the ideas of Ernst Mayr and Theodosius Dobzhansky, who argued 70 years ago that Darwin was wrong about species. Here, I show how recent genetic studies of supposedly well-behaved animals, such as insects and vertebrates, including our own species, have supported the existence of the Darwinian continuum between varieties and species. Below the level of species, there are well-defined ecological races, while above the level of species, hybridization still occurs, and may often lead to introgression and, sometimes, hybrid speciation. This continuum is evident, not only across vast geographical regions, but also locally in sympatry. The existence of this continuum provides good evidence for gradual evolution of species from ecological races and biotypes, to hybridizing species and, ultimately, to species that no longer cross. Continuity between varieties and species not only provides an excellent argument against creationism, but also gives insight into the process of speciation. The lack of a hiatus between species and ecological races suggests that speciation may occur, perhaps frequently, in sympatry, and the abundant intermediate stages suggest that it is happening all around us. Speciation is easy!     

Evolution rampant: house mice on Madeira

House mice are extra-ordinary animals – extra -ordinary in the literal sense of that word. They are pests – but also a valued laboratory animal. They are generalized rodents – and successful in habitats from tundra to tropics and from sea-level to high altitudes. They have differentiated into a perplexity of taxa, yet differ little in their general morphology. They were long scorned by ecologists as recently arrived commensals, but are increasingly illuminating evolutionary processes as new techniques are applied to their study. Local forms, once valued only by taxonomists, are proving ever more interesting as their genetics are probed. In 1992, Mathias & Mira described the apparently unexciting characteristics of mice living on the two main islands of the Madeira group, 600 km west of continental Portugal. Then in 2000, Britton-Davidian et al. discovered that there were at least six chromosomal (Robertsonian) races on the main island. In the past decade, studies of molecular and mitochondrial genomes have shown an array of variables and posed questions about the origins and subsequent evolution of these island mice. In this issue of Molecular Ecology , Förster et al. report on the mtDNA haplotypes found on the island and in mainland Portugal, discuss the probable source of the island colonizers, and consider data which might give information about the timing of the colonizing event(s).     

Meeting review: American Genetics Association Symposium on the genetics of speciation

Yeast can be engineered to carry human chromosomes; highly diverged ducks can produce viable, fertile offspring; and mitochondrial genes can move between widely divergent groups of plants. Some sunflower or oak species have porous genomes; mice, crickets, birds, and butterflies form hybrid zones; and bacterial lineages have been exchanging genes for several billion years. Even so, nature is discrete and full of species. Here, we discuss some of the ingredients that make nature discrete and can lead to clustering even in the presence of gene flow. Many of these results have been recently published, in this issue and elsewhere, and were discussed at the Genetics of Speciation Symposium held at the annual meeting of the American Genetics Association, Vancouver, Canada, in 2006.     

Robertsonian translocations in free-living populations of the house mouse in Belgium

Mice with Robertsonian (Rb) translocations have been discovered in some 45 populations in Europe. In Belgium, we have observed either Rb(4.12) in a heterozygous or homozygous state (2 n = 39 and 38 respectively), or Rb(4.12) homozygous with a heterozygous Rb(5.10), or both fusions in a homozygous state (2 n = 37 and 36, respectively). Some locations are highly polymorphic and hétérozygotes are found frequently. These observations suggest that the Rb population in Belgium could be of recent origin and probably is the result of introgression from Alsace (France) or Germany.