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Bandicoot rats (genus Bandicota), widely known as rodent pests, are abundant and widespread throughout the continental part of the Indo-Malayan realm. However, their evolutionary history is not yet well understood. The molecular phylogenetic relationships of the three bandicoot rat species, Bandicota bengalensis, Bandicota indica, and Bandicota savilei, were assessed based on the gene sequences of the specimens collected from Myanmar, where all three species occur along with database sequences. Early divergence of B. savilei (1.5–1.7 million years ago) was inferred from the mitochondrial cytochrome b (Cytb) gene and the nuclear interphotoreceptor retinoid-binding protein (Irbp), and melanocortin 1 receptor (Mc1r) gene sequences. The Cytb lineage of B. bengalensis from Sri Lanka was distinct from the monophyletic lineage of the continental lineages of B. bengalensis and B. indica. This can be explained by the preservation of ancient mitochondrial DNA (mtDNA) in the insular population owing to female philopatry and male dispersal, given that no substantial intraspecies geographic subdivision was observed in the nuclear markers. The paraphyletic relationship of B. bengalensis with B. indica may be explained by introgression of the mtDNA from B. bengalensis to B. indica, but further investigation is required to confirm this. B. bengalensis Cytb sequences from a wide area of Myanmar had limited nucleotide diversity (π = 0.00079), implying that the genetic diversity of B. bengalensis in Myanmar was acquired through Holocene human activities.

The Japanese archipelago is comprised of four main islands—Hokkaido, Honshu, Shikoku, and Kyushu—which contain high mountainous areas that likely allowed for lineage differentiation and population genetic structuring during the climatic changes of the late Pleistocene. Here, we assess the historical background of the evolutionary dynamics of herbivorous red-backed voles (Myodes) in Japan, examining the evolutionary trends of mitochondrial cytochrome b gene (Cytb) sequence variation. Four apparent signals from rapid expansion events were detected in three species, M. rufocanus and M. rutilus from Hokkaido and M. smithii from central Honshu. Taken together with results from previous studies on Japanese wood mice (Apodemus spp.), three of the expansion events were considered to be associated with predicted bottleneck events at the marine isotope stage (MIS) 4 period, in which glaciers are thought to have expanded extensively, especially at higher elevations. In the late Pleistocene, the possible candidates are transitions MIS 6/5, MIS 4/3, and MIS 2/1, which can be characterized by the cold periods of the penultimate glacial maximum, MIS 4, and the last glacial maximum, respectively. Our data further reveal the genetic footprints of repeated range expansion and contraction in the northern and southern lineages of the vole species currently found in central Honshu, namely M. andersoni and M. smithii, in response to climatic oscillation during the late Pleistocene. The time-dependent evolutionary rates of the mitochondrial Cytb presented here would provide a possible way for assessing population dynamics of cricetid rodents responding to the late Pleistocene environmental fluctuation.

Hantaviruses are harbored by multiple small mammal species in Asia, Europe, Africa, and the Americas. To ascertain the geographic distribution and virus-host relationships of rodent-borne hantaviruses in Japan, Vietnam, Myanmar, and Madagascar, RNAlater™-preserved lung tissues of 981 rodents representing 40 species, collected in 2011–2017, were analyzed for hantavirus RNA by RT-PCR. Our data showed Hantaan orthohantavirus Da Bie Shan strain in the Chinese white-bellied rat (Niviventer confucianus) in Vietnam, Thailand, orthohantavirus Anjo strain in the black rat (Rattus rattus) in Madagascar, and Puumala orthohantavirus Hokkaido strain in the grey-sided vole (Myodes rufocanus) in Japan. The Hokkaido strain of Puumala virus was also detected in the large Japanese field mouse (Apodemus speciosus) and small Japanese field mouse (Apodemus argenteus), with evidence of host-switching as determined by co-phylogeny mapping.

Additional file 3: Figure S2. A possible link between the population dynamics of Kyushu moles and late Quaternary environmental fluctuations. a An MJ network of the Cytb sequence dataset of moles from Kyushu and two southern peripheral islands of Tanegashima and Yakushima, showing two star-shaped clusters, termed Mwo-IIIa and Mwo-IIIa-1, indicative of rapid expansion events that are predicted to have occurred ca. 11,600 (Stage a) and ca. 15,000 (Stage b) years ago, respectively. b A schematic representation of the expanded land mass during the last glacial period approximately 16,000 years ago, when the sea level was ~ 120 m lower than at present [50]. The broad line shows the estimated coastline at the last glacial maximum [49]. c Global sea level change for the last 35,000 years, covering the last glacial maximum (LGM) and Younger Dryas (YD) [51, 52]. Arrows indicate the presumed time points of Stages a and b, when the two rapid expansion events of the mitochondrial gene haplogroups of Mwo-IIIa-1 and Mwo-IIIa are thought to have initiated in the mainland of Kyushu and the region that also included the two islands, respectively.

Additional file 1: Table S1. Specimens of Mogera imaizumii, Mogera wogura, and Mogera robusta used for molecular analyses in this study.

Additional file 2: Figure S1. a. The marine oxygen isotope curve over the last 150,000 years, adapted from Lisiecki and Raymo (2005) [51], with indication of the marine isotope stage (MIS). The four critical periods for small mammal population dynamics in Japan and the nearest continental areas are shown. Abrupt warmings after substantially cold periods are marked with letters (a, b, c, d) and boxes of different colors; those immediately after the end of the Younger Dryas (YD; a, light blue), the end of the last glacial period (b, dark blue), the early MIS 3 (c, green), and MIS 5e (d, orange) [3, 7, 8]. b The detailed climatic fluctuations (source: climate.gov/sites/default/files/default/files/historictemperaturerecord_greenland_large.jpg , download 20 May 15) are shown with the two prominent time periods: YD (a) and LGM (b). c-f Plots of the τ values obtained from previous studies of Apodemus speciosus (c), A. argentesus (d), Myodes voles (e) [3, 7, 8], and the present study of Mogera moles (f). The geographic map shows the approximate distribution of each haplotype group.

The Japanese archipelago is comprised of four main islands—Hokkaido, Honshu, Shikoku, and Kyushu—which contain high mountainous areas that likely allowed for lineage differentiation and population genetic structuring during the climatic changes of the late Pleistocene. Here, we assess the historical background of the evolutionary dynamics of herbivorous red-backed voles (Myodes) in Japan, examining the evolutionary trends of mitochondrial cytochrome b gene (Cytb) sequence variation. Four apparent signals from rapid expansion events were detected in three species, M. rufocanus and M. rutilus from Hokkaido and M. smithii from central Honshu. Taken together with results from previous studies on Japanese wood mice (Apodemus spp.), three of the expansion events were considered to be associated with predicted bottleneck events at the marine isotope stage (MIS) 4 period, in which glaciers are thought to have expanded extensively, especially at higher elevations. In the late Pleistocene, the possible candidates are transitions MIS 6/5, MIS 4/3, and MIS 2/1, which can be characterized by the cold periods of the penultimate glacial maximum, MIS 4, and the last glacial maximum, respectively. Our data further reveal the genetic footprints of repeated range expansion and contraction in the northern and southern lineages of the vole species currently found in central Honshu, namely M. andersoni and M. smithii, in response to climatic oscillation during the late Pleistocene. The time-dependent evolutionary rates of the mitochondrial Cytb presented here would provide a possible way for assessing population dynamics of cricetid rodents responding to the late Pleistocene environmental fluctuation.

The discovery of highly divergent lineages of hantaviruses (family Hantaviridae) in shrews, moles, and bats of multiple species raises the possibility that non-rodent hosts may have played a significant role in their evolutionary history. To further investigate this prospect, total RNA was extracted from RNAlater&reg, preserved lung tissues of 277 bats (representing five families, 14 genera and 40 species), captured in Myanmar and Vietnam during 2013&ndash, 2016. Hantavirus RNA was detected in two of 15 black-bearded tomb bats (Taphozous melanopogon) and two of 26 Pomona roundleaf bats (Hipposideros pomona) in Myanmar, and in three of six ashy leaf-nosed bats (Hipposideros cineraceus) in Vietnam. Pair-wise alignment and comparison of coding regions of the S, M, and L segments of hantaviruses from Taphozous and Hipposideros bats revealed high nucleotide and amino acid sequence similarities to prototype L&aacute, ibīn virus (LAIV) and Xu&acirc, n Sơn virus (XSV), respectively. Phylogenetic analyses, generated by maximum-likelihood and Bayesian methods, showed a geographic clustering of LAIV strains from China and Myanmar, but not of XSV strains from China and Vietnam. These findings confirm that the black-bearded tomb bat is the natural reservoir of LAIV, and that more than one species of Hipposideros bats can host XSV.

A phylogenetic tree was reconstructed based on the mitochondrial cytochrome b gene nucleotide sequences of 169 individuals of house shrews (Suncus murinus and S. montanus) from 44 localities in East Asia, Southeast Asia, West Asia, and islands in the western Indian Ocean. Shrews from China (Zhejiang), Japan (Okinawa), Vietnam, and Indonesia (Java) formed a monophyletic group with less genetic variation. Therefore, the shrews of these regions appeared to have originated from one or a few localities. Contrary to this, shrews from Sri Lanka, Myanmar, and Pakistan consisted of several haplogroups. This finding suggests immigration movements to these areas. Fascinating findings were also obtained concerning the islands in the western Indian Ocean. First, shrews on Zanzibar Island (Tanzania) had almost the same haplotype as those in southwestern Iran. Therefore, the house shrew in Zanzibar may have immigrated from Iran (or vice versa). Second, shrews from Madagascar and Grande Comore Island shared the same haplotype, whereas the shrews on Reunion Island were clearly different from those of Madagascar and Comoros. Thus, there appears to have been several immigration routes to the islands of the western Indian Ocean.

Variability in the coat color of the house mouse, Mus musculus, provides an opportunity to study the evolution of phenotypes in this species. Here we associated genetic variations with coat color in seven mice from Madagascar that had identical M. m. gentilulus mitochondrial DNA sequences. The entire coding region of the 948-base pair (bp) coat-color-related gene, Mc1r, was shown to have no nonsynonymous changes. However, analyses of the two exon-1 promoter regions—termed 1A (317 bp) and 1B (499 bp)—from a second gene, Asip, which is also involved in the evolution of coat color, revealed two distinct haplotypes in each region. Associations between Asip promoter regions and dorsal color were ambiguous; however, two ventral color types—light and dark gray—were associated with the haplotypes of 1A, as determined by clustering analysis. Notably, the haplotype of the light gray animals was identical to the Asip Aw allele that is associated with white bellies.

We assessed dispersal and vicariant events in four species of Japanese moles in the genera Mogera and Euroscaptor to better understand the factors shaping intra- and interspecific differentiation in Japanese moles. We used the combined viewpoints of molecular phylogeny and historical geology using nucleotide sequences of mitochondrial (cytochrome b; Cytb) and nuclear (A2ab, Bmp4, Tcf25, vWf) genes. The divergence times estimated from the molecular data were verified with available geological data on the chronology of fluctuations in sea level in the Korea Strait, assuming sequential migration and speciation events. This produced possible migration times of 5.6, 3.5, 2.4, and 1.3 million years ago for four species of Japanese moles, Euroscaptor mizura, Mogera tokudae, M. imaizumii, and M. wogura, respectively. For the western Japanese mole M. wogura, Cytb sequences revealed four major phylogroups with strong geographic affinities in southwestern Central Honshu (I), western Honshu/Shikoku (II), Kyushu/westernmost Honshu (III), and Korea/Russian Primorye (IV). The nuclear gene sequences supported the distinctiveness of phylogroups I and IV, indicating long, independent evolutionary histories. In contrast, phylogroups II and III were merged into a single geographic group based on the nuclear gene data. Intraspecific divergences in M. imaizumii and M. tokudae were rather apparent in Cytb but not in nuclear gene sequences. The results suggest that repeated dispersal events have occurred between the Asian continent and the Japanese Islands, and intensive vicariant events associated with abiotic and biotic factors have created higher levels of species and genetic diversities in moles occurring on the Japanese Islands.

We previously revealed the presence of six genetically distinct matrilineal populations of the Japanese dormouse Glirulus japonicus in the distribution range of Honshu, Shikoku, and Kyushu islands. In this study, we extended this analysis using mitochondrial cytochrome b gene sequences (n = 96) and Y-chromosome-specific SRY gene sequences (n = 22) from individuals collected from Honshu, Shikoku, Kyushu, and Oki Dogo I. The cytochrome b sequence data allowed us to define precise geographic ranges of the six previously known and three newly found distinct matrilineal lineages: northeastern Honshu (I), east-central Honshu (II), west-central Honshu and the Kii Peninsula (III), the western part of Honshu (IV), Shikoku (V), westernmost Honshu and Kyushu (VI), the northern part of central Honshu (VII), the southern part of central Honshu (VIII), and Oki Dogo I. (IX). Our inference of geographic borders suggests that regions of lower and higher altitudes in the mountain systems played important roles in driving the hosting and separation of lineages, respectively. Six matrilineal lineages (I, II, V, VI, VIII, and XI) were shown to possess their own SRY haplotypes, while lineages III and IV shared one haplotype. These data together with our previous observation of nuclear ribosomal RNA gene variation indicate advanced populational subdivision in this species. It is thus evident that each of the populations, including those living at high latitudes and in limited geographic spaces, have survived for several million years. A specific ability to tolerate cold may have permitted G. japonicus to preserve anciently diverged lineages in each locality.

A phylogeographic study of the large Japanese wood mouse, Apodemus speciosus, on Sado Island, Japan, was performed based on sequences of the mitochondrial cytochrome b gene (1,140 bp). Our previous study covered the entire species range and suggested that the mice on Sado Island are monophyletic, exhibiting two well diverged lineages throughout the island. The present data also supported two lineages (the average number of nucleotide difference was 11.4), showing a weak phylogeographic structure. Given the high sequence divergence observed, we assumed historically subdivided populations within the island. Bayesian coalescent analysis supported a dual-population model rather than that of one large population. The times to most recent common ancestor of all sequences were 293,000 years ago [ka; 95% highest probability density (HPD) 85-634 ka] and 292 ka (HPD 102-605 ka) for the one- and dual-population models, respectively. These results suggest that the populations have undergone repeated separations and reconnections, rather than being subdivided completely through time. Our results are in accordance with other paleogeographic and phylogeographic evidence from the island. The present study highlighted a unique system of producing and maintaining genetic diversity and suggested prehistoric colonization of the A. speciosus population on Sado Island, thus supporting the ancient origin of the mammalian fauna of Sado Island.

Three novel families of repetitive DNA sequences were molecularly cloned from the Korean field mouse (Apodemus peninsulae) and characterized by chromosome in-situ hybridization and filter hybridization. They were all localized to the centromeric regions of all autosomes and categorized into major satellite DNA, type I minor, and type II minor repetitive sequences. The type II minor repetitive sequence also hybridized interspersedly in the non-centromeric regions. The major satellite DNA sequence, which consisted of 30 bp elements, was organized in tandem arrays and constituted the majority of centromeric heterochromatin. Three families of repetitive sequences hybridized with B chromosomes in different patterns, suggesting that the B chromosomes of A. peninsulae were derived from A chromosomes and that the three repetitive sequences were amplified independently on each B chromosome. The minor repetitive sequences are present in the genomes of the other seven Apodemus species. In contrast, the major satellite DNA sequences that had a low sequence homology are present only in a few species. These results suggest that the major satellite DNA was amplified with base substitution in A. peninsulae after the divergence of the genus Apodemus from the common ancestor and that the B chromosomes of A. peninsulae might have a species-specific origin.

Skulls of the three species of Ryukyu spiny rats (Tokudaia osimensis, T. tokunoshimensis and T. muenninki) were osteometrically compared. In all measurements the size tendencies in which T. tokunoshimensis was the largest, T. muenninki middle, and T. osimensis the smallest were confirmed. However, the lengths of the molar row were exceptionally the largest in T. muenninki, middle in T. tokunoshimensis and the smallest in T. osimensis. The raw measurement data obviously indicated significant differences in size among the three species. We demonstrated that the neurocranium was larger in T. osimensis than in T. muenninki from the results of the proportional analyses. The principal component analyses using the raw measurement data also pointed out that the skulls from the three species were obviously distinguished in the charts, although the plots were not completely separated by the principal component analysis using the proportional indices. The CT images showed that both maxilla and palatine bone...

Comparative chromosome painting with mouse (Mus musculus, MMU) chromosome-specific DNA probes was performed for three Muridae species, the Indian soft-furred field rat (Millardia meltada), the spiny mouse (Acomys dimidiatus) and the harvest mouse (Micromys minutus). All probes except for the Y probe were successfully hybridized to the chromosomes of all species, and homologous chromosome segments between mouse and the three species were identified at the molecular level. Comparison of our data with the published data of six other genera (Mus, Rattus, Apodemus, Otomys, Rhabdomys and Cricetulus) of the Muridae suggested that the associations MMU1b/17a, 2b/13a, 5b/11a, 7/19, 10b/17b, 10c/17c, 11b/16a, 12/17d and 13b/15, and the single painted chromosomes and chromosome segments MMU3, 4, 5a, 8a, 8b, 16b, 18 and X were probably contained by the ancestral karyotype of the Muridae, and have been strongly conserved throughout murid evolution.

We evaluated the endothelial function of thoracic aortas and pulmonary arteries in a population of European wood mice (Apodemus sylvaticus), which exhibit hypercholesterolemia. According to the plasma cholesterol level, mice were divided into two groups: hypercholesterolemic (AHL, total plasma cholesterol 200-300 mg/dl) and normocholesterolemic (ANL, total plasma cholesterol

The sequences of the mitochondrial cytochrome b gene and restriction site variation in the spacer region of the nuclear ribosomal RNA gene [rDNA-restriction fragment length polymorphism (RFLP)] were analysed to determine the phylogeographic structure of the Japanese dormouse (Glirulus japonicus), which is threatened by deforestation and has been designated an endangered species in Japan. The phylogenetic tree of cytochrome b grouped G. japonicus into six geographical populations: north-eastern Honshu (I), central Honshu (II), west-central Honshu/Kii Peninsula (III), western Honshu (IV), Shikoku (V), and westernmost Honshu/Kyushu (VI); the genetic distances among these groups suggest divergence in the Late Tertiary. The lineage of group VI was located at the basal position in the phylogenetic tree, followed by the radiation of the other lineages. An rDNA-RFLP analysis of 15 restriction sites roughly supported such genetic isolation; groups I, II, III, IV, V and VI have five, two, one, one, one and four unique restriction sites, respectively, revealing four geographic groups as cryptic species: I, II, III + IV + V and VI. Our results reveal the ancient divergences of the local population, which has a complicated evolutionary history, and should be useful in developing a framework for the conservation of this species. Resumen Secuencias del gen mitocondrial citocromo b y variaciones en sitios de restriccion en la region espaciadora del gen nuclear de ARN ribosomal [rARN-fragmentos de restriccion de polimorfismos de longitud (RFLP)] fueron analizados para determinar la estructura filogeografica del liron japones (Glirulus japonicus), el cual es amenazado por deforestacion y ha sido designado como especie en peligro de extincion en Japon. El arbol filogenetico usando el gen mitocondrial citocromo b revelo que individuos de las especie G. japonicus estan divididos en seis regiones geograficas: (I) el Noreste de la isla de Honshu, (II) el centro de la isla de Honshu, (III) el oeste central de la isla de Honshu/la peninsula de Kii, (IV) el oeste de la isla de Honshu, (V) la isla de Shikoku y (VI) la region mas al oeste de las islas de Honshu y Kyushu. Las distancias geneticas entre estos grupos sugieren que la divergencia entre ellos ocurrio durante el Terciario tardio. El grupo VI ocupo el punto mas basal del arbol filogenetico, seguida de la radiacion de los otros grupos. Nuestro analisis de rADN-RFLP de 15 sitios de restriccion corroboraron los resultados del gen mitocondrial respecto al aislamiento genetico: los grupos I, II, III, IV, V y VI poseen cinco, dos, uno, uno, uno y cuatro sitios de restriccion unicos, respectivamente. Estos resultados permitieron reconocer cuatro grupos geograficos, I, II, III + IV + V y VI como una especie criptica. Los datos presentados aca revelan la divergencia ancestral de las poblaciones de Japon, las cuales presentan una complicada historia evolutiva. Estos datos seran de gran valor al momento de desarrollar estrategias de conservacion de esta especie.

House mouse (Mus musculus) is one of the perilous animal vectors for imported zoonosis such as a lymphocytic choriomeningitis (LCMV) infectious disease, and probably unknown emerging and/or re-emerging infectious diseases as well. It is necessary to prevent such diseases by regular surveys for behavioral trends of these allochthonous mice. However, such a trial has never been attempted in Japan. From 1998 to 2002, we analyzed partial sequences of the D-loop region in mtDNA, which provides powerful diagnostic SNPs for subspecies identification in the Mus musculus species, from 301 individuals of mice collected in 23 international bays or airports in Japan. We found that invasion of many allochthonous mice, which were identified as European subspecies, Mus musculus domesticus, occurred in Tokyo metropolitan coastal area. Based on the evidence, we warn that extensive invasion of allochthonous mice has occurred recently and, therefore, the risk of emerging and/or re-emerging infectious diseases invasion might be high in Tokyo metropolitan area.

Hybrid sterility plays an important role in the maintenance of species identity and promotion of speciation. Male interspecific hybrids from crosses between Campbell's dwarf hamster (Phodopus campbelli) and the Djungarian hamster (P. sungorus) exhibit sterility with abnormal spermatogenesis. However, the meiotic phenotype of these hybrids has not been well described. In the present work, we observed the accumulation of spermatocytes and apoptosis of spermatocyte-like cells in the testes of hybrids between P. campbelli females and P. sungorus males. In hybrid spermatocytes, a high frequency of asynapsis of X and Y chromosomes during the pachytene-like stage and dissociation of these chromosomes during metaphase I (MI) was observed. No autosomal univalency was observed during pachytene-like and MI stages in the hybrids; however, a low frequency of synapsis between autosomes and X or Y chromosomes, interlocking and partial synapsis between autosomal pairs and γ-H2AFX staining in autosomal chromatin was observed during the pachytene-like stage. Degenerated MI-like nuclei were frequently observed in the hybrids. Most of the spermatozoa in hybrid epididymides exhibited head malformation. These results indicate that the pairing of X and Y chromosomes is more adversely affected than that of autosomes in Phodopus hybrids.

The lesser Japanese mole, Mogera imaizumii, recognized by Motokawa and Abe (1996), occurs in eastern Honshu, western Honshu and Shikoku. Mitochondrial and nuclear DNA were analyzed for mole samples from eastern Honshu to elucidate intraspecific differentiation. Analyses of sequences of the mitochondrial cytochrome b gene (Cytb) and of a restriction fragment length polymorphism of the nuclear 28S ribosomal RNA gene spacer (rDNA-RFLP) revealed two genetic types, partially corresponding to Hutterer's (1993) taxa, M. wogura (= M. imaizumii) and M. minor. Most samples showed either of two combinations of mitochondrial / nuclear gene types. However, two specimens showed a different combination. This incongruent combination of mitochondrial and nuclear genes might have derived, in part, from an introgression event between genetically differentiated populations after secondary contact during the evolutionary history of the lesser Japanese mole in eastern Honshu.

A new species of Ryukyu spiny rat, Tokudaia tokunoshimensis was described in the specimens originating from Tokunoshima Island in the southernmost region of Japan. The populations of Tokudaia are separately distributed only in Amami-Oshima, Tokunoshima, and Okinawa-jima Islands. We have described the osteological and external morphological characteristics and clarified the osteometrical distinctions among the three populations using the skull and skin collections. All external dimensions of head and body length, tail length, hindfoot length and ear length were larger in the Tokunoshima population than in the other two in mean value. The raw osteometrical data separated the Tokunoshima Island population from the other two in all measurements except for the length of molar row in the Okinawa-jima Island population. The principal component analysis also demonstrated that the skulls from the Tokunoshima Island population were morphologically distinct from those of the other populations. Here, from th...

We examined intraspecies genetic variation in house mice (Mus musculus molossinus) from the northern third of the Japanese Islands, in order to obtain evidence of the history of mouse colonization that might have shaped the current genetic diversity. We extended the previous sampling of mitochondrial cytochrome b sequence and added information from the Y-linked Sry gene and ribosomal RNA gene surveys. We distinguish mitochondrial haplotypes characteristic of the North Asian musculus subspecies group (involving M. m. musculus and M. m. molossinus) as 'MUS', and that of the Southeast Asian castaneus subspecies group as 'CAS' (although the mice resemble MUS morphologically). There was a clear geographic partition of MUS and CAS types into southern and northern Hokkaido, respectively. Conversely, on Tohoku, the MUS and CAS types were interspersed without clear geographic subdivision. In contrast to the mtDNA data, all Hokkaido and Tohoku mice examined were found to possess a unique type for the Y-linked Sry gene, specific to Korea and Japan. Restriction site analysis of nuclear rDNA probe showed a consistent distribution of MUS and CAS types, as major and minor components, respectively, in the Hokkaido and Tohoku mice. These data support the previous notion that the Hokkaido and Tohoku mice experienced genetic hybridization between primary residents of CAS origin and MUS newcomers arriving via a southern route. The invasion of the MUS type could correspond with the evidence for arrival of prehistoric peoples. There are, however, alternative interpretations, including genetic admixture between MUS arriving by a southern route and CAS from a northern route.

We examined sequence variation in the mitochondrial cytochrome b gene (1140 bp, n = 73) and control region (842–851 bp, n = 74) in the Eurasian harvest mouse (Micromys minutus (Pallas, 1771)), with samples drawn from across its range, from Western Europe to Japan. Phylogeographic analyses revealed region-specific haplotype groupings combined with overall low levels of inter-regional genetic divergence. Despite the enormous intervening distance, European and East Asian samples showed a net nucleotide divergence of only 0.36%. Based on an evolutionary rate for the cytochrome b gene of 2.4%·(site·lineage·million years)–1, the initial divergence time of these populations is estimated at around 80 000 years before present. Our findings are consistent with available fossil evidence that has recorded repeated cycles of extinction and recolonization of Europe by M. minutus through the Quaternary. The molecular data further suggest that recolonization occurred from refugia in the Central to East Asian region. Japanese haplotypes of M. minutus, with the exception of those from Tsushima Is., show limited nucleotide diversity (0.15%) compared with those found on the adjacent Korean Peninsula. This finding suggests recent colonization of the Japanese Archipelago, probably around the last glacial period, followed by rapid population growth.

Factors affecting body temperature (Tb) fluctuations in winter were examined in Korean field mice Apodemus peninsulae. Animals were maintained under the following three conditions to mimic autumn followed by winter, plus two modes of behaviour, solitary and grouped, in winter: 1) short day (8 Light:16 Dark) at 25 ± 2°C and solitary, for 12 weeks towards the end of which their Tb was recorded; 2) constant darkness (DD) at 4 ± 2°C and solitary, for a week; and 3) DD at 4 ± 2°C and in groups (two or three animals in a cage), for about 10 days. Using implanted data loggers Tb was monitored in 13 mice that were exposed to these three conditions in sequence. When the animals' body temperature was below 30°C, it was assumed that they were torpid according to the bimodal pattern of minimum daily Tb. The maximum period of daily torpor was 390 min/day, and torpor was exhibited by seven of the thirteen animals exposed to conditions-2) and -3). Since torpor was found in one mouse even under condition-1), wit...

We sequenced the cytochrome b gene from two little-studied mammal species from the highlands of Southwest China, the long-tailed mole Scaptonyx fusicaudus and the gracile shrew-like mole Uropsilus gracilis. This data was used to examine the phylogenetic relationships among 19 talpid species within the family Talpidae (Mammalia: Eulipotyphla). Cytochrome b gene trees supported a basal placement of shrew-like moles (Uropsilus) within the Talpidae, and suggested that fossorial specializations arose twice during talpid evolution. To assess the evolutionary relationships of moles endemic to this region, we additionally sequenced the 12S rRNA gene and the nuclear recombination-activating gene-1 from eight and ten East Asian taxa, respectively. Analyses of these single and concatenated data sets suggested that East Asian shrew moles diverged prior to the evolution of fossorial Eurasian moles. However, we were unable to determine whether semi-fossorial shrew moles are monophyletic. In contrast, fossorial Eurasian genera (Talpa, Mogera and Euroscaptor) were consistently found to form a monophyletic clade, with Mogera and Euroscaptor representing sister taxa. Furthermore, this fossorial clade grouped with the semi-aquatic Desmana, although with fairly low (35-62%) bootstrap support. Mogera imaizumii was found to be more closely related to M. wogura than to M. tokudae. This implies that the ancestors of these three species entered Japan from the Asian continent in this order via a series of migration events, suggesting that the Japanese Islands have played an important role in preserving mole lineages from ancient to recent times.

Sequences of the mitochondrial cytochrome b (1140 bp) and nuclear IRBP (1152 bp) genes were used to assess the evolutionary history of Apodemus, using the complete set of Asian species. Our results indicate that speciation in Asia involved three radiations, which supports an earlier study. The initial radiation yielded A. argenteus (Japanese endemic), A. gurkha (Nepalese endemic), and the ancestral lineage of the remaining Asian species. This lineage subsequently diverged into four groups: agrarius-chevrieri (agrarius group), draco-latronum-semotus (draco group), A. peninsulae, and A. speciosus (Japanese endemic). The final step consisted of divergence within two species groups as a consequence of the geography of the Yunnan-Guizhou plateau and Taiwan. The ecological ability of two Apodemus species to inhabit one locality via niche partitioning likely drove the second radiation and shaped the basic geographical pattern seen today: A. argenteus and A. speciosus in Japan, A. agrarius and A. peninsulae in northern China, and the A. agrarius and A. draco groups in southern China. The three radiations are estimated to have occurred 7.5, 6.6, and 1.8–0.8 Mya respectively, using the IRBP clock, based on rat–mouse divergence 12 Mya. © 2003 The Linnean Society of London, Biological Journal of the Linnean Society, 2003, 80, 469–481.

Peroxidase-labeled staphylococcal protein A, streptococcal protein G, and antibodies directed against Mus musculus (mouse), Rattus norvegicus (rat), Mesocretus auratus (hamster), and Peromyscus leucopus were examined for their reactivity with immunoglobulin G (IgG) from various rodent species. The purpose of this study was to identify the optimal secondary antibodies or reagents for specific serodiagnosis of hantavirus infection in various rodent species. Using ELISA, a total of 65 sera from 29 rodent species of the family Muridae and one serum sample from family Octodontidae were compared for IgG reactivity with the six different reagents. The results demonstrate that the reactivities of the secondary antibodies and reagents to the sera varied, even among sera from rodents of the same genus. Hantavirus-specific antibody ELISA revealed that hantavirus-infected rodent sera obtained from M. musculus, R. norvegicus, Apodemus agrarius, A. peninsulae, and Bandicota indica bound to the six different conjugates in a similar pattern as that detected in IgG ELISA. These results indicate that the applicability of secondary antibodies and protein A and G should be carefully evaluated before use for serodiagnosis in different rodent species.

We conducted a polymerase chain reaction--restriction fragment length polymorphism (PCR-RFLP) analysis of the mitochondrial cytochrome b gene to elucidate the detailed genetic population structure of Japanese wild populations of medaka, Oryzias latipes. The analysis of 1,225 specimens collected from 303 sites identified 67 mitotypes. Subsequently we determined the nucleotide sequences of the complete cytochrome b gene (1141-bp) to clarify the phylogenetic relationships among mitotypes. The phylogenetic tree based on nucleotide sequences indicated three major clades (A, B and C) that differed by 11.3-11.8%, corresponding to three clusters previously identified by RFLP analysis of entire mitochondrial DNAs. The geographic distribution of mitotypes in clades A and B was fully concordant with the Northern and Southern Populations defined by allozymes. Clade A could be subdivided into three subclades and clade B into eleven, with sequence divergences among subclades of 1.3-5.8%. Each distribution of mitotypes in subclades roughly corresponded to that of mtDNA haplotypes in subclusters previously identified. Mitotypes in clade C were found only in the Kanto district. The phylogenetic relationships and the estimated divergence times suggest that three Japanese clades originated from a common ancestor and were separated during the Pliocene, and that the regional differentiation of subclades was closely connected with the geological history of the Quaternary. This study has also demonstrated the possibility of artificial disturbance of natural distribution especially in the Kanto district and the superior efficacy of PCR-RFLP analysis as a simple method for detecting genetic variation and artificial gene flow of medaka.

Experimental infection with Fasciola hepatica and parthenogenetic Fasciola sp. in laboratory animals have been conducted in rats and rabbits. Inoculation of less than 5 metacercariae into rat-like hamsters, Tscherskia triton, is sufficient to establish Fasciola infections. The prepatent period of F. hepatica and the parthenogenetic Fasciola sp. in T. triton was shorter than that in rats and rabbits, suggesting that T. triton is a suitable experimental model for these flukes. In contrast, F. gigantica infection in T. triton did not yield adult flukes; T. triton, is therefore, considered to be an unsuitable host for F. gigantica. The cotton rat, Sigmodon hispidus, was an unsuitable host for the parthenogenetic Fasciola sp.

Phylogenetic relationships among 20 species-group taxa of Mustelidae, representing Mustelinae (Mustela, Martes, Gulo), Lutrinae (Enhydra), and Melinae (Meles), were examined using nucleotide sequences of the nuclear interphotoreceptor retinoid binding protein (IRBP) and mitochondrial cytochrome b genes. Neighbor-joining and maximum-parsimony phylogenetic analyses on these genes separately and combined were conducted. While IRBP performed better than cytochrome b in recovering more-inclusive clades, cytochrome b demonstrated more resolving power in recovering less-inclusive clades. Strong support was found for a close affinity of Enhydra with Mustela to the exclusion of Martes and Gulo (causing Mustelinae to be paraphyletic); the most-basal position of Mustela vison within Mustela, followed by Mustela erminea; an association of Mustela lutreola, Mustela itatsi, Mustela sibirica, and the subgenus Putorius (including Mustela putorius and Mustela eversmanii), to the exclusion of Mustela nivalis and Mustela altaica; and a basal position of Mustela itatsi to a clade containing Mustela sibirica and Putorius. Whereas cytochrome b strongly supported Mustela lutreola as the sister species to Putorius, IRBP strongly supported its basal placement to the Mustela itatsi-Mustela sibirica-Putorius clade. The low level of sequence divergence in cytochrome b between Mustela lutreola and Putorius is therefore a result of interspecific mitochondrial introgression between these taxa, rather than a recent origin of Mustela lutreola in a close relationship to Putorius. Time estimates inferred from IRBP and cytochrome b for mustelid divergence events are mostly in agreement with the fossil record.

Comparative chromosome painting was applied to the Indian spiny mouse (Mus platythrix) with mouse (M. musculus) chromosome-specific probes for understanding the process of chromosome rearrangements between the two species. The chromosome locations of the 5S and 18S-28S ribosomal RNA genes and the order of the 119 and Tcp-1 genes in the In(17)2 region of the t-complex were also compared. All the painting probes were successfully hybridized to the Indian spiny mouse chromosomes, and a total of 27 segments homologous to mouse chromosomes were identified. The comparative FISH analysis revealed that tandem fusions were major events in the chromosome evolution of the Indian spiny mouse. In addition, other types of chromosome rearrangements, i.e. reciprocal translocations and insertions, were also included.

The parotid and mandibular glands of the cotton rat were examined by light and transmission electron microscopy. Parotid gland: Acinar cells were serous in nature, and contained electron-dense granules. Intercalated duct cells contained electron-dense granules. Striated duct cells had small granules of moderate and high electron densities. Mandibular gland: Acinar cells were seromucous in nature, and contained granules of low and moderate electron densities. Intercalated duct cells contained granules of moderate and high electron densities. Striated ducts were comprised of two portions - a secretory portion and a striated portion without granules. The secretory portion had many electron-dense granules. A sexual dimorphism was obserbed in these granules, which were smaller and fewer in females than in males.

We examined intraspecific genetic variation in the mitochondrial cytochrome b gene (1140 bp) in the Okinawa mouse, Mus caroli (Rodentia, Muridae), from Okinawa, Taiwan, Hainan, Yunnan (southern China), Thailand, Vietnam and Java, to better understand their evolutionary history and human impact on dispersal and colonization by the mice. Populations from each of the seven localities had distinctive mitochondrial DNA sequences. The genetic distances, ranging from 0.015 to 0.045 (0.033 on average), were comparable with inter-subspecies differences in the house mouse Mus musculus (0.024). Assuming that nucleotide substitutions occur at a constant rate of 0.024/million years/lineage for mitochondrial DNA, the divergence of in M. caroli is estimated to have occurred 0.3-0.9 million years ago. The relatively great extent of intraspecific mitochondrial DNA variation in this taxon, and the spatial mode of variation suggest that M. caroli is genetically structured in space, to a considerable extent. Our data imply that local insular populations of M. caroli have been separated for long evolutionary periods, and that the scattered distribution patterns in the insular domains are due to historical shrinkage of appropriate areas of habitat, rather than to recent establishment of dispersed local populations by inadvertent human introductions.

Molecular phylogeny of crocidurine shrews (Insectivora, Soricidae) in northeastern Asia was investigated to confirm the taxonomic status of unidentified specimens of Crocidura from Cheju Island, South Korea. Phylogenetic trees were constructed by neighbor-joining (NJ) and maximum likelihood (ML) methods, based on the mitochondrial cytochrome b gene sequences (402 base pairs) of 40 individuals of seven crocidurine species and three unidentified specimens from 31 localities mainly in northeastern Asia. Phylogenetic positions of the three unidentified specimens from Cheju Island were compared with those of Suncus murinus, C. attenuata, C. dsinezumi, C. lasiura, C. sibirica, C. suaveolens and C. watasei. Both in NJ and ML trees, the three unidentified specimens were included in the cluster of C. dsinezumi and obviously different from C. suaveolens on Cheju Island. Thus, the present investigation demonstrated that both of C. suaveolens and C. dsinezumi exist on Cheju Island.

Apodemus peninsulae is a field mouse that inhabits the broad-leafed forests of temperate Eurasia. We examined the mitochondrial cytochrome b gene in 57 individuals of A. peninsulae from northeastern Asia, including Siberia, Primorye, Magadan region, Sakhalin, Hokkaido, and the Korean Peninsula. The genealogy of the mitochondrial DNA (mtDNA) in A. peninsulae was shown to have substantial geographic affinity, suggesting geographic architecture of northeastern Asia, including the islands of Sakhalin and Hokkaido, played important roles on the cladogenesis. Taking into account the presence of region-specific anciently divergent mtDNA types, three parts of the regions of Primorye, Siberia, and the Korean Peninsula can be denoted as refugia for A. peninsulae during the substantial period of the Quaternary glacial ages. Among the geographic regions examined, Primorye is likely to be the most influential one, from which the mtDNA is thought to have migrated to the neighboring regions of Sakhalin, Hokkaido, the Magadan region, and Siberia during the evolution of this species.

Karyotypes of the greater Japanese shrew-mole, Urotrichus talpoides (Talpidae), collected from 57 localities in Japan were studied by conventional, G- and C-band staining techniques to elucidate geographical chromosomal variations, the mechanism of changes in chromosomes, and the distribution patterns of karyotypic races. Shrew-mole samples examined could be divided geographically into two slightly divergent chromosomal forms designated as the western and eastern races. These two karyotypic races had the same diploid number (34) and fundamental number (64), but they had autosomes with different makeups. A comparison of conventional karyotypes showed a distinct intraspecific variation in shape of autosomal pair no. 14 which was classified as subtelocentric in the western race and as metacentric in the eastern race. G- and C-banding analyses revealed that karyotypic variation found in no. 14 pair was involved in pericentric inversion and quantitative changes in constitutive heterochromatin. Intrasp...

We constructed the comparative cytogenetic maps of X chromosomes in three rodent species, Indian spiny mouse (Mus platythrix), Syrian hamster and Chinese hamster, using 26 mouse cDNA clones. Twenty-six, 22 and 22 out of the 26 genes, which were mapped to human, mouse and rat X chromosomes in our previous study, were newly localized to X chromosomes of Indian spiny mouse, and Syrian and Chinese hamsters, respectively. The order of the genes aligned on the long arm of human X chromosome was highly conserved in rat and the three rodent species except mouse. The present results suggest a possibility that the rat X chromosome retains the ancestral form of the rodent X chromosomes.

We constructed comparative cytogenetic maps of the Chinese hamster to mouse, rat and human by fluorescence in-situ hybridization using 36 cDNA clones of mouse, rat, Syrian hamster, Chinese hamster and human functional genes. In this study, 30 out of the 36 genes were newly mapped to Chinese hamster chromosomes. The chromosomal homology of the Chinese hamster was identified and arranged in 19, 19 and 18 segments of conserved synteny in mouse, rat and human, respectively. Additionally, two of the 19 segments homologous to mouse chromosomes were initially identified in this study.

To identify alpha(1)-adrenergic receptor subtypes involved in the regulation of cerebral microcirculation, we studied alpha(1)-adrenergic receptor subtypes expressed in the rat cerebral microvessels. The microvessels were prepared from rat cerebral cortex by albumin flotation and glass bead filtration techniques. [(125)I]HEAT binding to the cerebral microvessels was displaced by low concentrations of 5-methylurapidil, a selective antagonist for alpha(1A)-receptors, and modified Scatchard analysis of the data revealed that half of alpha(1)-receptors is alpha(1A)-subtype, and that alpha(1B)- and/or alpha(1D)-receptors are also present. The K(i) value of the high-affinity component for 5-methylurapidil was 3.90+/-1.08 nM, which is comparable with the value obtained in the rat cerebral cortex (2.17+/-0.88 nM). Reverse transcription and polymerase chain reaction experiments showed that mRNAs of alpha(1A)- and alpha(1B)-receptors, but not alpha(1D)-receptors, were expressed in the cerebral microvessels. These results suggest that alpha(1)-receptors involved in the regulation of cerebral microvessel function are alpha(1A)- and alpha(1B)-receptor subtypes.

Diplothrix legata and Tokudaia osimensis are rodent species endemic to the Ryukyu Islands (Okinawa area) of the southern part of Japan. To place them within a phylogenetic framework of Asian rodent species, we examined the molecular relationships among these species and Apodemus, Micromys, Mus, and Rattus, using an analysis of mitochondrial cytochrome b gene (1140 bp) and nuclear IRBP gene (782 bp) sequences. The results indicate that the four main murine genera and Tokudaia diverged at similar evolutionary times, namely at the time of the radiation of Murinae, 14 to 40 million years ago, depending on different concepts for the divergence times of Rattus and Mus. In contrast, Diplothrix separated from the Rattus group more recently. Our results indicate that D. legata and the Rattus group diverged at 20–30% of the divergence time of Rattus and Mus under the assumption of the molecular clock, suggesting that D. legata established its lineage at least by the beginning of the Pleistocene era. It is thus evident that the Okinawa area preserves rare indigenous species with various levels of genetic endemicity.

Species of field mice (genus Apodemus) are the most common rodents inhabiting woodlands and forests of the Palaearctic region. We examined the cytochrome b (cyt b) gene in mitochondrial DNA (1140 bp) and the interphotoreceptor retinoid binding protein (IRBP) gene in nuclear DNA (1152 bp) in nine species of Apodemus. Based on the genetic variation, the nine species were grouped into four lineages: (1) Agrarius group (A. agrarius, A. peninsulae, A. semotus, and A. speciosus), (2) Argenteus group (A. argenteus), (3) Gurkha group (A. gurkha), and (4) Sylvaticus group (A. alpicola, A. flavicollis, and A. sylvaticus). It was shown that these four lineages diverged within a short period of evolutionary time, suggestive of a radiation event. Soon after the radiation, the Agrarius group was likely to have differentiated again into the species lineages simultaneously. In contrast, the European clade, the Sylvaticus group, radiated rather recently. The relative ratio of the extent of sequence divergence among the four main lineages to that among the members of the subfamily Murinae (including Mus and Rattus) was calculated to be 72.4% in the cyt b gene with transversional substitutions, and 58.5% in the IRBP gene with all substitutions, using the Kimura two-parameter method. The value for the three European lineages was 27.6% in the cyt b gene and 12.3% in the IRBP gene. These results may have a correlation with the notion that deciduous broadleaf forests remained in Central East Asia through the late Tertiary to the present, while those in Europe to a large extent had disappeared by the Pliocene.