Microsatellite DNA markers indicate three genetic lineages in East Asian indigenous goat populations.

The genetic differentiation and phylogenetic relationships of 18 indigenous goat populations from seven East Asian countries were analysed based on data obtained from 26 microsatellite DNA markers. The mean number of alleles (MNA) per population ranged from 2.5 to 7.6, with an average of 5.8. Genetic variability estimated from MNA and heterozygosity (H(E) and H(O) ) were relatively low in coastal and island populations. A heterozygous deficiency within populations (F(IS) = 0.054, P < 0.001) and total inbreeding (F(IT) = 0.181, P < 0.01) were observed, and genetic differentiation in the populations (F(ST) ) was 13.4%. The results of Bayesian model-based clustering and a neighbour-joining tree based on Nei's genetic distance showed that Asian goat populations could be subdivided into at least the following three genetic clusters: East Asian, Southeast Asian and Mongolian. These results are in close accordance with conventional morphological and geographical classifications and migration history.

A method for discriminating a Japanese chicken, the Nagoya breed, using microsatellite markers.

The Nagoya breed native to Japan is popular as a dual-purpose breed for eggs and meat. The current study describes a method to discriminate between the Nagoya breed and other breeds and commercial stocks of chicken. Four strains of the Nagoya breed established at the Aichi-ken Agricultural Research Center were analyzed using 25 microsatellite markers. In these strains, 5 of the markers (ABR0015, ABR0257, ABR0417, ABR0495, and ADL0262) had a single allele. Other chicken samples (448) of various breeds and hybrids were analyzed using the same 5 markers. None of these chicken samples had the same allele combination as the Nagoya breed strains. These 5 microsatellite markers provide a practical method to accurately discriminate the Nagoya breed from other chicken breeds.

A chicken linkage map based on microsatellite markers genotyped on a Japanese Large Game and White Leghorn cross.

A detailed linkage map is necessary for efficient detection of quantitative trait loci (QTL) in chicken resource populations. In this study, microsatellite markers isolated from a (CA)n-enriched library (designated as ABR Markers) were mapped using a population developed from a cross between Japanese Game and White Leghorn chickens. In total, 296 markers including 193 ABR, 43 MCW, 31 ADL, 22 LEI, 3 HUJ, 2 GCT, 1 UMA and 1 ROS were mapped by linkage to chicken chromosomes 1-14, 17-21, 23, 24, 26-28 and Z. In addition, five markers were assigned to the map based on the chicken draft genomic sequence, bringing the total number of markers on the map to 301. The resulting linkage map will contribute to QTL mapping in chicken.

A linkage map of 243 DNA markers in an intercross of Göttingen miniature and Meishan pigs.

A resource family of pigs has been constructed by using a boar of Göttingen miniature pig and two sows of Meishan pig as parents. In the construction of the family, two F1 males and 18 F1 females were intercrossed to generate 143 F2 offspring. The members of the family were genotyped using 243 genetic markers including 26 markers developed in our laboratory in order to generate a linkage map of markers for use in detecting quantitative trait loci (QTLs) in the family. The markers consisted of 237 microsatellites, five PRE-1 markers, and one RFLP marker. The linkage map was revealed to cover all 18 autosomes and the X chromosome; and the total length of the sex-averaged linkage map was calculated to be 2561.9 CM. Four out of the 26 markers developed in our laboratory exended the current linkage map at the termini of chromosomes 1p, 5p, 11p, and Xq. The linkage maps of all the chromosomes except for chromosome 1 were found to be longer in females than in males. Concerning chromosome 1, the length of the linkage map showed no difference between females and males, which was attributed to low recombination rates between markers localized in the centromeric region in females. The average ratio of female-to-male recombination was calculated to be 1.55.

The pig aminoacylase 1 (ACY1) and ribosomal protein L29 (RPL29)/heparin/heparan sulfate interacting protein (HIP) genes are located together at 13q21-->q22, corresponding to human 3p21.1.

The pig aminoacylase 1 (ACY1; N-acylamino acid aminohydrolase) gene was isolated from a pig cosmid library and characterized. The gene spans about 4.7 kb and consists of 15 exons. Fluorescence in situ hybridization found ACY1 to be located on pig chromosome 13 in the region q21-->q22. This result and previous reports show that a large part of pig chromosome 13 corresponds to human chromosome 3. BLAST search results reveal that chromosome 13 contains a transcript similar to human ribosomal protein L29 (RPL29)/heparan sulfate/heparin-interacting protein (HIP). The transcript lies near the 3'-flanking region of the pig ACY1 gene; the 2 genes are linked tail-to-tail. The deduced amino acid sequence shows distinct homology to human RPL29/HIP, 96% identical in the N-terminal region. In the corresponding human 3p21.1 region, a deletion closely linked to the ACY1 locus has been observed in carcinoma cells. This suggests that a tumor suppressor gene is located in this region. Comparative mapping suggests also that the human RPL29/HIP gene may be near ACY1. Because many growth factors and cytokines interact with cells via heparin/heparan sulfate-proteoglycan, RPL29/HIP may play an important role on the cell surface by modulating interactions between cells and extracellular molecules.

Freemartinism among singleton bovine females born from multiple embryo transfer.

Heterosexual chimerism among singleton females produced by multiple nonsexed embryo transfer (MNET singleton females) was investigated using chromosome typing and PCR (polymerase chain reaction)-amplification of male-specific DNA (msDNA). Of the 22 animals tested, 21 were classified as normal by both methods (i.e., showing no male cells among 100 metaphase spreads in chromosome typing and being msDNA negative in PCR). No morphological abnormalities of the genital organs were observed among 19 MNET single females. One MNET singleton female was, however, classified as a freemartin by PCR (male-specific DNA positive), but it was classified as normal cytogenetically. This individual probably had a low degree of heterosexual chimerism, and it seems that the chimerism derived from MNET was difficult to diagnose by chromosome typing, although it was detectable by PCR. The genital organs of this individual (15-mo-old Aberdeen Angus) were normal in form (both external and internal) and size. However, a very small structure, resembling seminiferous tubule, was found in the left ovary. It may be concluded that most MNET singleton females are expected to have normal reproductive function.

Recovery of dissected C-band regions in Crepis chromosomes.

Chromosome samples were prepared on a plastic coverslip covered with a polyester membrane and were subjected to the C-banding treatment. The C-band pattern was obtained after Giemsa staining. The C-band positive regions of the Crepis chromosomes were identified, dissected out by irradiation with a micro-laser beam and recovered in Eppendorf tubes.

Measurement of serum IgE antibodies against Japanese cedar pollen (Cryptomeria japonica) in Japanese monkeys (Macaca fuscata) with pollinosis.

IgE antibodies against allergens of Japanese cedar (Cryptomeria japonica, CJ) pollen in the serum of seven Japanese monkeys (Macaca fuscata) with pollinosis were measured by fluorometric indirect enzyme-linked immunosorbent assay (ELISA). All of the monkeys were found to have specific IgE to the crude pollen antigen. The specific IgE levels were well correlated with those determined by the Pharmacia CAP system. IgE antibodies were then assayed with two kinds of purified allergens (Cry j I and Cry j II) by the ELISA. We found that five monkeys had specific IgE to both allergens, although the other two had IgE only to Cry j I or Cry j II; there is different immune responsiveness to the two major allergens in the monkeys.

Microdissection of plant chromosomes by argon-ion laser beam.

Rice and barley chromosomal samples were prepared both on a polyester membrane and on an ordinary glass slide and subjected to microdissection by an argon-ion laser. The intensity and the position of the laser beam were controlled by a microcomputer. The most suitable intensity to obtain chromosomal fragments was determined experimentally. As a result, specific regions of the centromere, satellite, short arm, or long arm, of the barley and rice chromosomes were dissected out from the chromosomal spreads. Chromosomal fragments were also successfully transferred from the sample into an Eppendorf tube.

Establishment of primate lymphoblastic cell lines by coculture with a simian T-cell leukemia virus-1 positive, hypoxanthine-guanine-phosphoribosyltransferase negative Japanese macaque cell line.

A cell line (JAMH17+) resistant to 8-azaguanine was established from a human T-cell leukemia virus type 1 related virus (simian T-cell leukemia virus-1) positive Japanese macaque cell line. Lymphoblastic cell lines were established from the peripheral blood mononuclear cells of humans, hominoids, and several species of macaques by coculture with JAMH17+ in hypoxanthine-aminopterin-thymidine medium. HTLV-1 specific antigen was detected in some of the established cell lines. Phenotypic analysis showed that several cell lines of crab-eating macaques expressed Leu11a antigen, which is a marker of human natural killer cells.

Allelic constitution of the hemoglobin beta chain in wild populations of the house mouse, Mus musculus.

We studied the allelic frequency of the hemoglobin beta chain (Hbb) of wild mice, Mus musculus, collected from 46 localities, mostly in Asia and surrounding areas. The wild populations in the northern part of China, Korea, and the central part of Japan exhibited an almost monomorphic distribution of Hbbp. In the southern part of Asia, the frequency of Hbbp decreased and Hbbd was predominant. Although Hbbs and Hbbd are generally found in Europe, the Hbbp allele was present in Southeastern Europe (Bulgaria). In the light of these results, the Hbbp allele might have originated in mice of northern Asia.

Isolation in culture of a type C virus from a Japanese monkey seropositive to adult T-cell leukemia-associated antigens.

Co-cultivation of lymphocytes from two Japanese monkeys, one of which was seropositive to adult T-cell leukemia (ATL)-associated antigens (ATLA), gave rise to a lymphoid cell line derived from the anti-ATLA negative monkey. This cell line harbors ATLA and type C virus particles identical in morphology to ATL virus.

Detection of type C virus particles in Japanese monkeys seropositive to adult T-cell leukemia-associated antigens.

Type C virus particles which were identical to adult T-cell leukemia virus (ATLV) in morphology were detected in phytohemagglutinin-stimulated peripheral lymphocytes from four out of four Japanese monkeys seropositive to adult T-cell leukemia-associated antigens. It is possible that non-human primates may be a natural reservoir of ATLV, and vector-borne transmission of ATLV from monkeys to humans should be explored.

Frequency distribution of histocompatibility-2 antigenic specificities in the Japanese wild mouse genetically remote from the European subspecies.

Fifty-eight Japanese wild mice, Mus musculus molossinus, collected from twenty-eight localities were surveyed for twelve H-2 antigens using the haemagglutination method. Significantly higher frequencies of H-2.3 and H-2.5 specifities and relatively lower frequencies of the other public specificities were observed. This was confirmed by examining four specificities, H-2.3, 5, 13 and 23, in 370 mice. Quantitative absorption of the alloantisera by erythrocytes and spleen lymphocytes of molossinus mouse revealed definite absorption of H-2.3, 5 and 8 antigens, though their antigenic strength was apparently weaker than the controls, B10 congenic mice. Comparative assay of the strength of H-2.5 antigen in either homozygous and heterozygous conditions distinctly eliminated the possibility of gene-dose effect for the reduced strength of the public antigens in molossinus mouse. To explain this, divergence time between molossinus and domesticus was computed based on the allelic frequency data already reported on ten loci in both subspecies. It is roughly 1.5 x 10(6) years, which could allow for the occurrence of considerable genetic changes in many public antigens, except those conserved through selection.