Mitochondrial DNA Sequence Variation in the Eastern House Mouse, Mus Musculus: Comparison with Other House Mice and Report of a 75-Bp Tandem Repeat

The control region and flanking tRNAs were sequenced from 139 Mus musculus mitochondrial DNAs (mtDNAs) from mice collected at 44 localities extending from Germany to Japan. Among the 36 types of M. musculus mtDNA resolved, five have an added 75-bp direct repeat; the two copies within an individual differ by two to four base substitutions. Among 90 M. domesticus mtDNAs sequenced, 12 new types were found; 96 M. domesticus types have now been identified by sequencing this segment. Representative mtDNAs from M. castaneus, M. macedonicus, M. spicilegus and M. spretus were also sequenced. A parsimony tree for the M. musculus mtDNAs is about half as deep as the tree for the M. domesticus mtDNAs, which is consistent with the idea that M. musculus is genetically less diverse and younger than M. domesticus. The patterns of variation as a function of position are similar but not identical in M. musculus and M. domesticus mtDNAs. M. castaneus and M. musculus mtDNAs are allied, at a tree depth about three times as great as the start of intra-M. musculus divergence. The coalescence of the M. musculus and M. castaneus mtDNAs is about half as deep as their coalescence with the M. domesticus mtDNA lineages. The mtDNAs of the aboriginal M. macedonicus and M. spicilegus are each other's closest relatives, at a tree depth greater than the deepest intracommensal node. The mtDNA results support the view that the aboriginal M. spretus is the sister group of the other five species.     

Genome Patterns of Selection and Introgression of Haplotypes in Natural Populations of the House Mouse (Mus musculus)

General parameters of selection, such as the frequency and strength of positive selection in natural populations or the role of introgression, are still insufficiently understood. The house mouse (Mus musculus) is a particularly well-suited model system to approach such questions, since it has a defined history of splits into subspecies and populations and since extensive genome information is available. We have used high-density single-nucleotide polymorphism (SNP) typing arrays to assess genomic patterns of positive selection and introgression of alleles in two natural populations of each of the subspecies M. m. domesticus and M. m. musculus. Applying different statistical procedures, we find a large number of regions subject to apparent selective sweeps, indicating frequent positive selection on rare alleles or novel mutations. Genes in the regions include well-studied imprinted loci (e.g. Plagl1/Zac1), homologues of human genes involved in adaptations (e.g. alpha-amylase genes) or in genetic diseases (e.g. Huntingtin and Parkin). Haplotype matching between the two subspecies reveals a large number of haplotypes that show patterns of introgression from specific populations of the respective other subspecies, with at least 10% of the genome being affected by partial or full introgression. Using neutral simulations for comparison, we find that the size and the fraction of introgressed haplotypes are not compatible with a pure migration or incomplete lineage sorting model. Hence, it appears that introgressed haplotypes can rise in frequency due to positive selection and thus can contribute to the adaptive genomic landscape of natural populations. Our data support the notion that natural genomes are subject to complex adaptive processes, including the introgression of haplotypes from other differentiated populations or species at a larger scale than previously assumed for animals. This implies that some of the admixture found in inbred strains of mice may also have a natural origin.     

Variation in Amylase Haplotypes among Congenic Lines of the House Mouse

Pancreatic amylase in the mouse displays considerable quantitative genetic variation. Agar gel electrophoresis reveals that homozygous animals have either one form of the enzyme, type A, or two forms, type AB. Only few animals have been found that contradict this statement, namely among Mus musculus castaneous from Thailand, which has a single-banded B type. Double-banded homozygous specimens of various origins have different relative proportions of the two isoenzymes. By measuring the A:B ratios in such animals, a number of distinct haplotypes or amylase complexes, determining ratios ranging from 61% A:39%, B to 12% A:88% B, have been recognized. These complexes differ also with respect to the total amount of amylase produced. If the reference stock C3H/As is given the value 1, then other haplotypes have values ranging from 1.0 to 0.27. Nineteen amylase haplotypes have been established in congenic lines on a C3H/As background. Some of these lines contain at least four active pancreatic amylase structural genes and breeding experiments have demonstrated that the genetic elements regulating total amylase production and relative proportions of the isoenzymes are located within the amylase complex, cis-acting, and very closely linked to the structural genes.     

Genetic and Taxonomic Diversity of the House Mouse Mus musculus from the Asian Part of the Former Soviet Union

Genetic diversity of the house mouse Mus musculus from 12 local populations (n = 65) of the central and eastern parts of the former Soviet Union was examined using RAPD–PCR. About 400 loci were identified, encompassing approximately 500 kb of the mouse genome. Genetic diversity was assessed using NTSYS, POPGENE, TFPGA, and TREECON software programs. In general, the house mouse sample from the regions examined was characterized by moderate genetic variation: polymorphism P = 95.6%, P ₉₉ = 60.7%, P ₉₅ = 24.2%; heterozygosity H = 0.089; the mean observed number of alleles n _a = 1.97; effective number of alleles n _e = 1.13; intrapopulation differentiation _S = 0.387; gene diversity h = 0.09. Individual local populations displayed different levels of genetic isolation: the genetic subdivision index G _st varied from 0.086 to 0.324 at gene flow Nm varying from 5.3 to 1.05, while the interpopulation genetic distance D _N ranged from 0.059 to 0.186. Most of the genetic diversity of the total sample resided within the local populations: H _S = 0.06, total gene diversity H _T = 0.09. The exact test for differentiation, however, did not confirm the affiliation of all the mice examined to one population: ² = 1446, d.f. = 724, P = 0.000. Molecular markers specific to four subspecies (musculus, castaneus, gansuensis, and wagneri) were identified. Moreover, in some cases the populations and individual animals exhibited traits of different subspecies, suggesting their introgressive hybridization. It was demonstrated that the house mouse fauna on the territories investigated was characterized by the prevalence of musculus-specific markers, while gansuensis-specific markers ranked second. The castaneus-specific markers were highly frequent in the Far East, but almost absent in Central Asia, where wagneri-specific markers were detected. It was suggested that house mice from Turkmenistan could belong to one of the southern subspecies, which had not deeply penetrated into the Asian fauna of the former Soviet Union. In phenogenetic (UPGMA) and phylogenetic (NJ) reconstructions this form with the high bootstrap support was placed at the tree base, while the isolation of other clusters was not statistically significant. It is thus likely that the house mice from Turkmenistan are closest to the ancestral form of the genus Mus on the territory of the former Soviet Union.     

Allozymic Polymorphism Among 14 Populations of the House Mouse, Mus musculus domesticus, From Greece

Nineteen loci from 239 individuals of the house mouse Mus musculus domesticus (Rodentia, Muridae) were analyzed by means of thin layer electrophoresis. The mice were collected from 14 localities of Greece mainly confined to the area of NW Peloponnese, where a Robertsonian (Rb) system is observed. The individuals were chromosomally characterized by nine diploid numbers, the 2n = 24, 26, 27, 28, 29, 30, 31, 32, and 40. The statistic elaboration revealed that all 14 populations studied were not characterized by cohesive demic structure and high inbreed levels while the gene flow among them has resulted in low levels of genetic differentiation. The resulting values for Neis genetic distance corresponded to distances known for the level of geographical populations of, M. musculus. Wagners cladogram for the phylogenetic relations between the populations studied implied that it is the diploid number, rather than the geographical factor, that characterizes or dominates each population, which mainly influences the phylogenetic relationships.     

Eradication of the House Mouse (Mus musculus) from Montague Island,New South Wales,Australia

Summary Of all the introduced mammalian pests, rodents are the most common invaders of islands and have triggered numerous extinctions around the globe. They have a short gestation period and large litter size, giving them the ability to colonise new areas rapidly. The same biological traits make them difficult to eradicate. Although the biodiversity benefits of removing exotic rodents from islands are increasingly being recognised, there are few published analyses of eradication attempts that critically evaluate eradication tools. This study examined the response of House Mouse (Mus musculus) populations to an eradication operation on Montague Island, Australia. While the specific impacts of mice have not been studied on this particular island, elsewhere they have been shown to have negative impacts on a range of species including plants, invertebrates, lizards and seabirds. On Montague Island, mouse abundance across different habitats was examined using mark–recapture data collected before and after the deployment of brodifacoum baits. Data obtained before baiting showed significantly lower numbers of mice in sites dominated by exotic grass compared with those dominated by native vegetation. Mouse numbers overall were declining during winter and the population was not breeding, making this an optimal time to undertake the eradication. Trapping immediately after the initial bait drop failed to capture any survivors, and no individuals have been detected during the 3 years since the deployment of baits. This study demonstrated that both small and large baits (0.6 and 2 g pellets) were effective in eradicating the House Mouse from Montague Island. While present, mice were probably slowing the re‐establishment of native vegetation by grazing seedlings and consuming seeds. With this pest now gone, the process of natural regeneration is expected to accelerate. The eradication of the House Mouse from Montague is likely to have other positive effects on the island’s biodiversity. The operation itself contributed to enhancing local capacity to eradicate exotic rodents from larger or more complex islands.     

Evidence for Resetting the Directional Component of Path Integration in the House Mouse (Mus musculus)

Path integration is the animal's running computation of its position relative to a starting point based on recording all displacements. This process is known to be prone to cumulative errors and hence must somehow be corrected with the help of local spatial cues. In the reported experiments, the relative role of local spatial cues at two target locations was appraised. These two targets were the peripheral box of a female house mouse (Mus musculus) and a central box from where she retrieved her pups over a distance of 50 cm. The experimental conditions required the mouse to navigate between the two targets solely by means of path integration. To find out whether the mice continued integrating at the two target locations, directional misinformation was fed into their path integrating system. Hence, passive rotation of 90° proved to be sufficient to decide unambiguously whether the mice were misdirected. Such experimental interference consistently documented ongoing path integration at the target location outside the residential nest. In contrast, the same interference when the mouse was in the residential nest documented discontinuation of path integration in most cases. It is inferred that mice, when departing from residential nests, initially direct themselves by means of local guiding stimuli.     

小鼠单个卵母细胞中18S rRNA基因表达研究

目的:探讨小鼠卵母细胞发育成熟与18S rRNA基因表达的关系。方法:根据Genbank中公布的小鼠18S rRNA保守区序列(363bp),用DNAclub软件设计两对特异性引物,RT-PCR检测单个GV、MⅠ期卵母细胞18S rRNA的表达;取单个MⅠ期卵母细胞二次PCR产物连接至pTG19-T载体、转化大肠杆菌感受态细胞,取阳性克隆进行测序。结果:RT-PCR检测显示18S rRNA基因在小鼠单个GV期、MⅠ期卵母细胞中均有表达,且在未成熟卵母细胞中,MⅠ期的表达明显强于GV期的表达。测序结果表明小鼠单个卵母细胞表达的18S rRNA基因与参考序列(Genbank NR_003278)完全一致。结论:小鼠MⅠ期之后的卵母细胞发育需要蛋白质的合成,卵母细胞的成熟可能与核糖体18S基因表达有关。     

Characterization of Triosephosphate Isomerase Mutants with Reduced Enzyme Activity in Mus Musculus

Four heterozygous triosephosphate isomerase (TPI) mutants with approximately 50% reduced activity in blood compared to wild type were detected in offspring of 1-ethyl-1-nitrosourea treated male mice. Breeding experiments displayed an autosomal, dominant mode of inheritance for the mutations. All mutations were found to be homozygous lethal at an early postimplantation stage of embryonic development, probably due to a total lack of TPI activity and consequently to the inability to utilize glucose as a source of metabolic energy. Although activity alteration was also found in liver, lung, kidney, spleen, heart, brain and muscle the TPI deficiency in heterozygotes has no influence on the following physiological traits: hematological parameters, plasma glucose, glucose consumption of blood cells, body weight and organo-somatic indices of liver, spleen, heart, kidney and lung. Biochemical investigations of TPI in the four mutant lines indicated no difference of physicochemical properties compared to the wild type. Results from immunoinactivation assays indicate that the decrease of enzyme activity corresponds to a decrease in the level of an immunologically active moiety. It is suggested that the mutations have affected the Tpi-1 structural locus and resulted in alleles which produce no detectable enzyme activity and no immunologically cross-reacting material. The study furthermore suggests one functional TPI gene per haploid genome in the erythrocyte and seven other tested organs of the mouse.     

A Novel Mouse Chromosome 17 Hybrid Sterility Locus: Implications for the Origin of T Haplotypes

The effects of heterospecific combinations of mouse chromosome 17 on male fertility and transmission ratio were investigated through a series of breeding studies. Animals were bred to carry complete chromosome 17 homologs, or portions thereof, from three different sources-Mus domesticus, Mus spretus and t haplotypes. These chromosome 17 combinations were analyzed for fertility within the context of a M. domesticus or M. spretus genetic background. Two new forms of hybrid sterility were identified. First, the heterospecific combination of M. spretus and t haplotype homologs leads to complete male sterility on both M. spretus and M. domesticus genetic backgrounds. This is an example of symmetrical hybrid sterility. Second, the presence of a single M. domesticus chromosome 17 homolog within a M. spretus background causes sterility, however, the same combination of chromosome 17 homologs does not cause sterility within the M. domesticus background. This is a case of asymmetrical hybrid sterility. Through an analysis of recombinant chromosomes, it was possible to map the M. domesticus, M. spretus and t haplotype alleles responsible for these two hybrid sterility phenotypes to the same novel locus (Hybrid sterility-4). Previous structural studies had led to the hypothesis that the ancestral t haplotype originated through an introgression event from M. spretus or a related species. If this were true, one might expect that (1) M. spretus homologs would be transmitted at a non-Mendelian ratio within the M. domesticus background, and (2) t haplotypes would be transmitted at a ratio closer to Mendelian within the M. spretus background.(ABSTRACT TRUNCATED AT 250 WORDS)     

Diversity of Extended HLA-DRB1 Haplotypes in the Finnish Population

The Major Histocompatibility Complex (MHC, 6p21) codes for traditional HLA and other host response related genes. The polymorphic HLA-DRB1 gene in MHC Class II has been associated with several complex diseases. In this study we focus on MHC haplotype structures in the Finnish population. We explore the variability of extended HLA-DRB1 haplotypes in relation to the other traditional HLA genes and a selected group of MHC class III genes. A total of 150 healthy Finnish individuals were included in the study. Subjects were genotyped for HLA alleles (HLA-A, -B, -DRB1, -DQB1, and -DPB1). The polymorphism of TNF, LTA, C4, BTNL2 and HLA-DRA genes was studied with 74 SNPs (single nucleotide polymorphism). The C4A and C4B gene copy numbers and a 2-bp silencing insertion at exon 29 in C4A gene were analysed with quantitative genomic realtime-PCR. The allele frequencies for each locus were calculated and haplotypes were constructed using both the traditional HLA alleles and SNP blocks. The most frequent Finnish A∼B∼DR -haplotype, uncommon in elsewhere in Europe, was A*03∼B*35∼DRB1*01∶01. The second most common haplotype was a common European ancestral haplotype AH 8.1 (A*01∼B*08∼DRB1*03∶01). Extended haplotypes containing HLA-B, TNF block, C4 and HLA-DPB1 strongly increased the number of HLA-DRB1 haplotypes showing variability in the extended HLA-DRB1 haplotype structures. On the contrary, BTNL2 block and HLA-DQB1 were more conserved showing linkage with the HLA-DRB1 alleles. We show that the use of HLA-DRB1 haplotypes rather than single HLA-DRB1 alleles is advantageous when studying the polymorphisms and LD patters of the MHC region. For disease association studies the HLA-DRB1 haplotypes with various MHC markers allows us to cluster haplotypes with functionally important gene variants such as C4 deficiency and cytokines TNF and LTA, and provides hypotheses for further assessment. Our study corroborates the importance of studying population-specific MHC haplotypes.     

Targeted Mutagenesis of a Candidate T Complex Responder Gene in Mouse T Haplotypes Does Not Eliminate Transmission Ratio Distortion

Transmission ratio distortion (TRD) associated with mouse t haplotypes causes +/t males to transmit the t-bearing chromosome to nearly all their offspring. Of the several genes involved in this phenomenon, the t complex responder (Tcr(t)) locus is absolutely essential for TRD to occur. A candidate Tcr(t) gene called Tcp10b(t) was previously cloned from the genetically defined Tcr(t) region. Its location, restricted expression in testis, and a unique postmeiotic alternative splicing pattern supported the idea that Tcp10b(t) was Tcr(t). To test this hypothesis in a functional assay, ES cells were derived from a viable partial t haplotype, and the Tcp10b(t) gene was mutated by homologous recombination. Mutant mice were mated to appropriate partial t haplotypes to determine whether the targeted chromosome exhibited transmission ratios characteristic of the responder. The results demonstrated that the targeted chromosome retained full responder activity. Hence, Tcp10b(t) does not appear to be Tcr(t). These and other observations necessitate a reevaluation of genetic mapping data and the actual nature of the responder.