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There is great uncertainty about how Ireland attained its current fauna and flora. Long-distance human-mediated colonization from southwestern Europe has been seen as a possible way that Ireland obtained many of its species; however, Britain has (surprisingly) been neglected as a source area for Ireland. The pygmy shrew has long been considered an illustrative model species, such that the uncertainty of the Irish colonization process has been dubbed 'the pygmy shrew syndrome'. Here, we used new genetic data consisting of 218 cytochrome (cyt) b sequences, 153 control region sequences, 17 Y-intron sequences and 335 microsatellite multilocus genotypes to distinguish between four possible hypotheses for the colonization of the British Isles, formulated in the context of previously published data. Cyt b sequences from western Europe were basal to those found in Ireland, but also to those found in the periphery of Britain and several offshore islands. Although the central cyt b haplotype in Ireland was found in northern Spain, we argue that it most likely occurred in Britain also, from where the pygmy shrew colonized Ireland as a human introduction during the Holocene. Y-intron and microsatellite data are consistent with this hypothesis, and the biological traits and distributional data of pygmy shrews argue against long-distance colonization from Spain. The compact starburst of the Irish cyt b expansion and the low genetic diversity across all markers strongly suggests a recent colonization. This detailed molecular study of the pygmy shrew provides a new perspective on an old colonization question.

Chromosomal races of the common shrew differ in sets of metacentric chromosomes and on contact may produce hybrids with extraordinarily complex configurations at meiosis I that are associated with reduced fertility. There is an expectation that these may be some of the most extreme tension zones available for study and therefore are of interest as potential sites for reproductive isolation. Here, we analyse one of these zones, between the Novosibirsk race (characterized by metacentrics go, hn, ik, jl, mp and qr) and the Tomsk race (metacentrics gk, hi, jl and mn and acrocentrics o, p, q and r), which form hybrids with a chain-of-nine (CIX) and a chain-of-three (CIII) configuration at meiosis I. At the Novosibirsk-Tomsk hybrid zone, the CIX chromosomes form clines of 8.53 km standardized width on average, whereas the cline for the CIII chromosomes was 52.83 km wide. The difference in these cline widths fits with the difference in meiotic errors expected with the CIX and CIII configuration, and we produce estimates of selection against hybrids with these types of configurations, which we relate to dispersal and age of the hybrid zone. The hybrid zone is located at the isocline at 200 m altitude above sea level; this relationship between the races and altitude is suggested at both coarse and fine scales. This indicates adaptive differences between the races that may in turn have been promoted by the chromosome differences. Thus, the extreme chromosomal divergence between the Novosibirsk and Tomsk may be associated with genic differentiation, but it is still striking that, despite the large chromosomal differences, reproductive isolation between the Novosibirsk and Tomsk races has not occurred., (© 2011 The Authors. Journal of Evolutionary Biology © 2011 European Society For Evolutionary Biology.)

The Moscow and Seliger chromosomal races of the common shrew differ by Robertsonian fusions and possibly whole-arm reciprocal translocations (WARTs) such that their F₁ hybrids produce a chain-of-eleven configuration at meiosis I and are expected to suffer substantial infertility. Of numerous hybrid zones that have been described in the common shrew, those between the Moscow and Seliger races involve the greatest chromosomal difference. We collected 211 individuals from this zone to generate a total dataset of 298 individuals from 187 unique global positioning system (GPS) locations within the vicinity of interracial contact. We used a geographic information system (GIS) to map the location of the hybrid zone, which follows a direct route between two lakes, as would be anticipated from tension zone theory. Even within the central area of the hybrid zone, there is a much higher frequency of pure race individuals than hybrid, making this a clear example of a bimodal zone in the sense of Jiggins & Mallet (2000). The zone runs through good habitat for common shrews, but nevertheless it is very narrow (standard cline widths: 3-4 km), as would be anticipated from low hybrid fitness. There is clear potential for an interruption to gene flow and build-up of reproductive isolation. As found in some other hybrid zones, there is a high frequency of novel genetic variants, in this case, new chromosomal rearrangements. Here, we report a de novo Robertsonian fission and a de novo reciprocal translocation, both for the first time in the common shrew. There is an extraordinarily high frequency of de novo mutations recorded in F₁ hybrids in the zone and we discuss how chromosomal instability may be associated with such hybrids. The occurrence of a de novo Robertsonian fission is of considerable significance because it provides missing evidence that fissions are the basis of the novel acrocentric forms found and apparently selected for in certain common shrew hybrid zones., (© 2010 The Authors. Journal of Evolutionary Biology © 2010 European Society For Evolutionary Biology.)

We investigated the evolution of the biomechanics of the mandible in island and mainland populations of the common shrew on the west coast of Scotland. We predicted that climatic differences between populations should cause differences in prey composition leading to changes in the mechanical potential (MP) of the mandible. In females, MP was correlated with climate, with greater MP in warmer and drier habitats. In males, MP was significantly greater than in females but there was no relationship between male MP and climate. This led to increased sexual dimorphism in colder and wetter climates. The same pattern was found after a phylogenetic least squares analysis was conducted to account for shared phylogenetic history. We discuss possible reasons for this pattern, including male-male combat and the greater necessity of females to feed as efficiently as possible to meet their extremely high energy requirements during lactation.

Mandibles from 13 island and six mainland populations of common shrews from the west coast of Scotland were subjected to geometric morphometric analysis in order to investigate the relationship between genetic diversity and fluctuating asymmetry. Although population mean shape fluctuating asymmetry (FA) and size FA were significantly inversely correlated with population genetic diversity this result was substantially due to one island. Sanda, the smallest island with by far the lowest genetic diversity, also had the highest FA. When Sanda was removed from the analysis, the relationship was not significant. There was no relationship between genetic diversity and FA at the individual level, whether measured as mean locus heterozygosity or d(2). In general, if genetic variation affects FA at all, the effect is weak and may only be of biological interest in very small populations.

A major problem in studies of synaptonemal complexes (SC) is the difficulty in distinguishing individual chromosomes. This problem can be solved combining SC immunostaining with FISH of chromosome-specific sequences. However, this procedure is expensive, time-consuming and applicable only to a very limited number of species. In this paper we show how a combination of SC immunostaining and DAPI staining can allow identification of all chromosome arms in surface-spreads of the SC of the common shrew (Sorex araneus L.). Enhancement of brightness and contrast of the images with photo editing software allowed us to reveal clear DAPI-positive and negative bands with relative sizes and positions similar to DAPI landmarks on mitotic metaphase chromosomes. Using FISH with DNA probes prepared from chromosome arms m and n we demonstrated correct recognition of the chromosomes mp and hn on the basis of their DAPI pattern. We show that the approach we describe here may be applied to other species and can provide an important tool for identification of individual bivalents in pachytene surface-spreads.

Crucial to our understanding of chromosomal variation and evolution in mammals are detailed studies of chromosomal heterozygotes, with analyses of chromosomal segregation and chromosome-derived infertility. We studied segregation and fertility in hybrids between karyotypic races of the house musk shrew Suncus murinus. These individuals were heterozygous for up to five Robertsonian fusions (Rbs) and an insertion of heterochromatin in an autosome. All variant chromosomes showed Mendelian segregation and all Rbs segregated independently of each other in the progeny of double heterozygotes. Litter size in single and even multiple Rb heterozygotes was no smaller than that in the less fertile parental strain. The effects of genetic background were more important in determining litter size than Rb heterozygosity for the shrews that we examined.

There is a general perception that central and northern Europe were colonized by range expansion from Mediterranean refugia at the end of the last glaciation. Data from various species support this scenario, but we question its universality. Our mitochondrial DNA studies on three widespread species of small mammal suggest that colonization may have occurred from glacial refugia in central Europe-western Asia. The haplotypes on the Mediterranean peninsulae are distinctive from those found elsewhere. Rather than contributing to the postglacial colonization of Europe, Mediterranean populations of widespread small mammals may represent long-term isolates undergoing allopatric speciation. This could explain the high endemism of small mammals associated with the Mediterranean peninsulae.

A Robertsonian fusion polymorphism in the common shrew (Sorex araneus L.), first described in Academgorodok near Novosibirsk (western Siberia) in 1970-72, was re-examined in 1994-95. The polymorphism in the 1970s involved chromosome arm combinations go, jl, mp and qr, i.e. each of these combinations was present in both a metacentric and a twin-acrocentric state in the population at that time. The twin-acrocentric morph for go occurred at low frequency in 1970-72 and was not observed in 1994-95. The polymorphism for arm combinations jl, mp and qr was still observed in 1994-95 and there was no significant difference in metacentric/twin-acrocentric frequencies compared with the previous sample. This is the third well-documented example in which the chromosome polymorphism in the common shrew has been found to be unchanged over a period of 20+ years. Although the polymorphism for qr may be associated with a chromosomal hybrid zone with a cline centre 200 km away, there is no definitive explanation for the other polymorphisms.

A high-resolution G-banding technique was used to identify five metacentrics that characterize Suncus murinus from Sri Lanka. These metacentrics were shown to be the product of Robertsonian fusion of acrocentric chromosomes identical to those in the standard karyotype defined by M.B. Rogatcheva et al. Two of the metacentrics in the Sri Lankan shrews (Rb(10.12) and Rb(14.15)) were the same as those reported by C.H. Sam et al. in Malayan populations of S. murinus. This finding provides strong support for the suggestion of T.H. Yosida that metacentric-carrying shrews colonized Malaya from Sri Lanka and hybridized with individuals of standard karyotype, generating the Robertsonian polymorphism now observed. In addition to the Robertsonian variation in S. murinus, we have used our high resolution technique (G- and C-banding) to characterize variants on chromosome 7, the X chromosome, and the Y chromosome.

This report extends the genetic map of the common shrew (Sorex araneus) by use of a clone panel of shrew-Chinese hamster and shrew-mouse hybrid cells (Pack et al., 1995; Matyakhina et al., 1996). This set of hybrid clones made it possible to assign the shrew genes for isocitrate dehydrogenase 2 (IDH2), inorganic pyrophosphatase (PP), glutamicpyruvate transaminase (GPT), adenosine kinase (ADK), glucuronidase 2 (GUSB) and acid phosphatase 2 (ACP2) to chromosome ik; the genes for adenylate kinases 1 and 3 (AK1 and AK3) to chromosome af; the genes for glutamate-oxaloacetate transaminase 2 (GOT2), peptidase D (PEPD) and growth hormone (GH) to chromosome hn; the gene for phosphoglucomutase 2 (PGM2) to chromosome go, the gene for enolase 1 (ENO1) to chromosome ji, the gene for ornithine carbamoyl-transferase (OTC) to chromosome de, the gene for aminoacylase 1 (ACY1) to arm m (chromosome mp), the gene for glutamate-oxaloacetate transaminase 1 (GOT1) to arm q (chromosome qr). Thus, the genetic map of the common shrew now contains 33 genes and it is possible to compare the syntenic associations with other species.

This report extends the genetic map of the common shrew (Sorex araneus) by adding chromosome assignments for ten genes to the seven already mapped (Pack et al. 1995). A somatic cell hybrid panel was used for the mapping. The genes for peptidase A (PEPA) and isocitrate dehydrogenase-1 (IDH1) map to chromosome de; the genes for phosphoglucomutase-1 (PGM1), superoxide dismutase-1 (SOD1), and mannosephosphate isomerase (MPI) are located on chromosome af; the genes for nucleoside phosphorylase (NP) and glutathione reductase (GSR) are on chromosome ik; and the genes for peptidase S (PEPS), malic enzyme-1 (ME1), peptidase B (PEPB) are found on chromosomes jl, go, and mp respectively.

The chromosome localization of the gene encoding growth hormone (GH) was determined by Southern blotting of DNA obtained from a panel of common shrew x Chinese hamster and common shrew x mouse hybrid somatic cell clones using mink GH DNA as a probe. The GH gene was found to be localized on chromosome hn of the common shrew.

We selected the common shrew (Sorex araneus) to generate the first insectivore gene map. Shrew-Chinese hamster and shrew- mouse somatic cell hybrid cells were constructed. When the 119 shrew-rodent clones were characterized, only shrew chromosomes were found to have segregated. A panel of hybrid clones was selected for gene assignment. The genes for hypoxanthine phosphoribosyl transferase (HPRT), glucose-6- phosphate dehydrogenase (G6PD), and malate dehydrogenase 1 (MDH1) were assigned to shrew Chromosome (Chr) de [which is the product of a tandem fusion between the 'original' mammalian X Chromosome (Chr) and an autosome], the gene for adenosine deaminase (ADA) and 6-phosphogluconate dehydrogenase se (PGD) to Chromosome jl, the gene for thymidine kinase (TK) to Chromosome hn, and the gene for lactate dehydrogenase (LDHA) to chromosome ik. Further studies in progress.

Female common shrews in their first pregnancy were collected near Oxford (UK) from the hybrid zone between the Oxford and Hermitage races. These races differ by Robertsonian chromosomal rearrangements and both Robertsonian heterozygotes and homozygotes are found in the zone. The three homozygotes examined in this study had significantly more oocytes than the six heterozygotes. Among the heterozygotes, the number of oocytes tended to be lower in more severely heterozygous individuals; one double heterozygote was particularly depleted. Although there were, on average, 41% fewer primordial follicles in the ovaries of the heterozygotes than those of the homozygotes, there was no significant difference in the numbers of growing follicles between the karyotypic classes. These data suggest that Robertsonian heterozygotes may have a shorter reproductive lifespan than do homozygotes, but the numbers of follicles being recruited for ovulation at a particular instance during the fertile period does not appear to differ between homozygotes and heterozygotes. Morphological differences between the follicles of homozygotes and heterozygotes were not detected. Only 0.12% of healthy growing follicles were biovular.

Three hypotheses concerning potential genetic benefits of female multiple mating behaviour are evaluated for the common shrew. In a high-density population, many successful copulations took place between individuals estimated to be close relatives (e.g. full or half siblings). Juveniles resulting from such matings tended to be relatively small at weaning, and were generally less likely to survive to sexual maturity than more outbred individuals. Multiple paternity was discovered in eight of nine litters examined. The incidence of matings between close relatives, the cost of inbreeding, and the high incidence of multiple paternity presented are each consistent with the hypothesis that female multiple mating is a strategy to reduce inbreeding. That is, if females cannot always distinguish close kin, then they may copulate with several different males and so reduce the risk that all their offspring will be sired by a close relative.

Two chromosomal races of common shrews (Sorex araneus) were crossed in captivity to generate chain VII-forming complex Robertsonian heterozygotes. Meiosis and gametogenesis were studied in three male hybrids. Regular chain VII configurations were observed at both pachytene and diakinesis/metaphase I, although in many pachytene spreads the chain configuration was incomplete (the basis of this peculiarity is unknown). From metaphase II counts, the frequency of anaphase I nondisjunction in the complex heterozygotes was estimated to be 13%. Germ-cell death in the chain VII-forming complex heterozygotes was 22% greater than it was in controls, but this difference is unlikely to have greatly influenced the capacity of the heterozygotes to sire offspring. Thus, the fecundity of these complex heterozygous common shrews would probably have been only slightly reduced relative to homozygous or simple heterozygous shrews. These results call into question the generality of speciation models based on the presumed sterility of complex heterozygotes.

We have tested for the occurrence of multiple paternity in wild common shrews by karyotypic analysis and DNA-fingerprinting of five wild-caught females and their litters. Karyotypic data suggest that some litters were sired by more than one male, but provide no definitive evidence. By using DNA-fingerprinting, it was possible to establish that two males sired the litter of two females. The present report shows that multiple paternity is not a rare phenomenon in the common shrew and by using DNA-fingerprinting it is possible to assign individual offspring to different male parents even when none of the putative fathers are available for inspection.

Twenty-two adult male common shrews were collected from 5 sites in the vicinity of Oxford (UK) close to the zone of hybridization between two karyotypic races. The shrews were subdivided into 3 karyotypic categories: homozygotes, simple Robertsonian heterozygotes (which form one or more trivalents at prophase I of meiosis) and complex Robertsonian heterozygotes (which form a quadrivalent). The ratio of primary spermatocytes to round spermatids was determined from transverse sections of seminiferous tubules, to provide an indication of germ cell death. In no individual was there severe germ cells loss. Homozygotes had the highest mean spermatocyte: spermatid ratio and complex heterozygotes the lowest, but there was substantial individual variation and the differences were not significant. Complex heterozygotes also had a higher proportion of defective seminiferous tubules and lower testis weights than did other categories and it is reasonable to propose that, as a population, complex heterozygotes had reduced fitness relative to other categories on the basis of spermatogenic performance. However, there is no evidence from studies of spermatogenesis that simple Robertsonian heterozygotes are less fit than homozygotes.

A variable kidney esterase found in the common shrew has a trimeric structure as indicated by the phenotypes of heterozygotes. From substrate specificity and tissue distribution this enzyme appears to be homologous with esterase-6 of the mouse, also a trimeric enzyme. It is suggested that a trimeric esterase may have been a feature of early eutherian mammals.

We present the G-band karyotype of Sorex granarius (Miller, 1910). With minor exceptions, each of the acrocentric autosomes in this karyotype is completely homologous to one of the chromosome arms in the karyotype of the common shrew (Sorex araneus L., 1758). If, as is the simplest interpretation, the karyotypic evolution in S. araneus is by repeated Robertsonian fusion, the karyotype in S. granarius represents the ancestral condition. We urge breeding studies to establish whether the designation of S. granarius as a full species is justified.

Samples of common shrews with an Aberdeen race, Oxford race and Hermitage race karyotype and from the Oxford-Hermitage hybrid zone were screened for genotype at six polymorphic enzyme loci. The common allele at all loci was the same in all samples suggesting that the degree of genic divergence between the karyotypic races is not great. However, shrews of the Aberdeen race appear to be somewhat distinct. There were differences in allele frequencies, range of alleles and heterozygosities at the Mpi-1, Pgm-2 and Pgm-3 loci between the Aberdeen race samples and samples of other karyotypic categories. The allele frequency variation across the Oxford-Hermitage hybrid zone is not substantially linked with karyotype frequency change, but there is notable allele frequency variation between close sites of similar habitat, particularly at the Pgm-2 and Pgm-3 loci. This suggests that gene flow between close sites may be reduced sufficiently in some instances to allow allele frequency change by genetic drift. This may have implications for the mode of origin of karyotypic races of common shrew.

Pregnant female common shrews were collected from an area of Robertsonian polymorphism, involving five different arm combinations, around Oxford (England). The females and their fetuses were karyotyped, and the karyotypes of the sires were deduced. Ten pregnancies where at least one parent was known to be either a single or double Robertsonian heterozygote were available for analysis. From these pregnancies, upper and lower estimates of anaphase I nondisjunction arising from Robertsonian heterozygosity were calculated to be 1.0-2.5% per heterozygous arm combination and 1.5-3.7% per heterozygous parent. One trisomic fetus with retarded development was identified. This trisomy can be attributed to anaphase I nondisjunction of a trivalent in a female Robertsonian heterozygote.

A method for the systematic breeding of the common shrew (Sorex araneus) is described. By use of this method 20 litters of common shrew, conceived in the wild and in captivity, were reared to weaning. Successful fertilization was recorded for 74% of pairings and 68% of young known to have been born were reared to weaning. Breeding records indicate that common shrews will mate within a few days of pairing and that the duration of pregnancy and lactation is about 43 days.

XY/XYY sex-chromosome mosaicism was demonstrated in both bone marrow and germ cells of a wild adult common shrew. Secondary sexual characteristics were those of a normal male, but the testes were small, and the sperm count was only about 3% of normal. Most of the seminiferous tubule cross-sections examined revealed serious spermatogenic impairment and a reduced diameter. A range of sex-chromosome pairing configurations was observed in XYY primary spermatocytes, including configurations involving the X and both Y chromosomes in a linear or radial array. The presence of metaphase II (MII) spreads with an XY sex-chromosome complement indicated that XYY primary spermatocytes could contribute products to MII. Following Burgoyne (1979) and Burgoyne and Biddle (1980), a number of models of spermatocyte loss were tested. The data indicated that there was an association between the sex-chromosome complement of primary spermatocytes and their contribution to MII. The best fit to the observed MII frequency data was provided by a model which assumed that all XYY primary spermatocytes with a univalent Y chromosome and a high proportion of XYY primary spermatocytes with an unpaired X chromosome failed to contribute products to MII.

An adult male common shrew with an XXY sex chromosome constitution was found in a natural population. The external appearance of the animal and size of the accessory glands were normal but the testes were as small as those of immature shrews. Histological preparations of the testis revealed seminiferous tubules containing only one type of cell, probably Sertoli cells, and interstitial cell hyperplasia.

A Robertsonian karyotypic polymorphism in the common shrew in the Oxford area, first described in the 1950s, was re-examined. The polymorphism involves chromosome arm combinations kq, no and pr (characteristic of the Oxford karyotypic race), ko (characteristic of the Hermitage karyotypic race) and jl (found in both races). The polymorphism for jl was sporadic along a north-south transect through the Oxford area, with the frequency of the twin-acrocentric morph never exceeding 10%. The frequency of the Oxford race-specific metacentrics decreased and the frequency of the Hermitage race-specific metacentric ko increased from north to south along the transect. At a latitudinal grid reference of about 180 km, there was a high frequency of individuals with chromosome arms k, n, o and q in the ancestral acrocentric state. This was coincident with the area of occurrence of ko-kq and ko-no Oxford-Hermitage hybrids. Such hybrids are double Robertsonian heterozygotes with monobrachial homology and are likely to suffer reduced fertility in consequence. It is proposed that this is a source of selection against the monobrachial hybrids and hence results in an increase in frequency of the acrocentric morphs. This scheme goes some way to explain the clines of polymorphism for arm combinations kq, no and ko, but it is suggested that other selective factors are involved. It cannot explain the cline of polymorphism for pr, which is in general terms similar to that for kq and no, but is more shallow and centred further north.

Adult male common shrews, both Robertsonian heterozygotes and homozygotes, were collected from Oxford and elsewhere in Britain. In both simple Robertsonian heterozygotes and Robertsonian heterozygotes with monobrachial homology, regular chain configurations were observed at meiosis I; only 1-2% were incomplete such that univalents were observed. On the average, there was one chiasma per chromosome arm among those that displayed Robertsonian variation, including both chain configurations and bivalents. According to one hypothesis, a single chiasma per chromosome arm may facilitate proper disjunction of chain trivalents of simple Robertsonian heterozygotes. Based on metaphase II counts, anaphase I nondisjunction frequency can be estimated as 1.0% per heterozygous individual and 0.7% per heterozygous arm combination.