Single nucleotide polymorphisms of the KIT and KITLG genes in pigs

Here, we report the variability in the KIT tyrosine kinase receptor and its ligand KITLG genes by determining single nucleotide polymorphisms (SNPs) in 384 individuals including 11 pig breeds, two synthetic-line cross pigs, two cross breeds, and one Japanese wild boar. SNPs and indels within the coding sequence regions of KITLG and KIT and their 5'-flanking regions were detected by aligning sequences from eight pigs, and subsequently the SNPs were genotyped using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI TOF-MS). Principal component analysis using allele frequencies in the SNP locus showed a distant relationship between Asian and Euro-American pig groups, except for Berkshire and Tokyo X breeds. These breeds were located within the mid-portion of the distribution in the first principal component. The Hampshire breed was distant from the other pig groups on the axis of the second principal component. Haplotype frequencies that were deduced using non-synonymous substitutions of the KIT gene revealed the uniqueness of Landrace, Large White, Middle White, and three-way cross pigs (LWD) and of the Hampshire breed. On the other hand, the haplotypes of KITLG and KIT detected in the Berkshire breed were prevalent in Asian pig groups. This tendency is different from that observed in other Euro-American pig breeds.     

Production of asexual and sexual offspring in the triploid sexual planarian Dugesia ryukyuensis

Certain freshwater planarians reproduce asexually as well as sexually, and their chromosomal ploidies include polyploidy, aneuploidy and mixoploidy. Previously, we successfully performed an experiment in which a clonal population produced by asexual reproduction of the Dugesia ryukyuensis (OH strain) switched to the sexual mode of reproduction. Worms of this strain are triploid with a pericentric inversion on Chromosome 4. The worms were switched to sexual reproduction after being fed with sexually mature Bdellocephala brunnea, which is a sexually reproducing species. The resulting sexualized OH strain produced cocoons filled with several eggs. Two putative factors, Mendelian factor(s) and chromosomal control(s), have been proposed as determining the reproductive mode. The present study demonstrated that inbreeding of the resultant sexualized worms produced the following four types of offspring through sexual reproduction: diploid asexual worms, triploid asexual worms, diploid sexual worms and triploid sexual worms. The chromosomal mutation on Chromosome 4 was inherited by these offspring independent of their reproductive mode. These results provide two important pieces of information: (i) the putative genetic factor was not necessarily inherited in a Mendelian fashion; and (ii) the reproductive mode is not regulated by chromosomal changes such as polyploidy or chromosomal mutations. This suggests that asexuality in D. ryukyuensis is regulated by an unknown factor(s) other than a Mendelian factor or a chromosomal control.     

A rapid multiple immunofluorescence method for the simultaneous detection of CD20 and CD3 in canine and feline cytological samples

CD20 and CD3 are considered reliable markers for B and T cells, respectively. This study aimed to develop a rapid multiple immunofluorescence (RMIF) method for the detection of CD20 and CD3 on a single cytology slide. Air-dried smears were prepared using samples collected from dogs (n=26) and cats (n=6). Immunosignal detection using the newly developed method required 60 min. Clear immunosignals for CD20 and CD3 were detected in 24 of 26 samples in dogs and in all 6 cats. As the RMIF (CD20/CD3) method can detect markers of both B and T cells simultaneously on a single cytology smear, it would be an efficient tool for the immunophenotyping of canine and feline lymphoma samples.     

Tethered spinal cord related to caudal spinal dysraphism in a tailless Holstein calf

A rare dysraphic caudal spinal anomaly, or caudal agenesis, comprising a tethered spinal cord, was found in a tailless Holstein calf that presented ataxia and paresis with analgesia of the hind limbs. The gently and slimly tapered conus medullaris was poorly formed between S2 and S3 which indicated that it was lying more caudally. The caudal end of the filum terminale adhered to the inner periosteum of the vertebral arch at S4, which is compatible with tethering of the spinal cord. The dysraphic changes from the secondary neurulation error and the longitudinal deranged cord morphology that may have been caused by the caudad traction due to tethering were confirmed. This represents the first bovine case with definitive morphological confirmation.     

Mobility of the forearm skeleton in the Asiatic black (Ursus thibetanus), brown (U. arctos) and polar (U. maritimus) bears

In several primates and carnivores, pronation/supination angles of the forearm skeleton were examined, and it is thought that a larger angle is useful to acquire dexterous behaviors in feeding and/or life style, including climbing. In this study, the pronation/supination angles in Asiatic black, brown and polar bears were nondestructively examined. These specimens were classified as adult or non-adult. Three or four carcasses of each group of Asiatic black and brown bears were used for CT analysis, whereas only one adult polar bear was used. The forearms were positioned within the gantry of a CT scanner in both maximally supinated and pronated states. Extracted cross-sectional CT images of two positions were superimposed by overlapping the outlines of each ulna. The centroids of the radii were detected, and then the centroid of each radius and the midpoint of a line which connects between both ends of the surface of each radius facing the ulna, were connected by lines to measure the angle of rotation as an index of pronation/supination. In adult brown and polar bears, the angles were smaller as compared with the other groups (Asiatic black and non-adult brown bears). Asiatic black and non-adult brown bears can climb trees, whereas adult brown bears and polar bears cannot. This suggests that the pronation/supination angle is related to arboreal activity in Ursidae.     

Maintenance of pregnancy in ovariectomized Mongolian gerbils (Meriones unguiculatus)

Bilateral ovariectomy (Ovx) was carried out on day 20 of pregnancy in Mongolian gerbils ( Meriones unguiculatus ). The body weights of all groups tended to decrease on the day after the operation, and the decrease was significant in the group that was ovariectomized and given vehicle (Ovx + vehicle group). The body weight in this group never recovered until autopsy on day 24, which is normally 1 day before parturition. No fetuses survived to the time of autopsy in any of the animals of the Ovx + vehicle group. Daily administration of 4 mg of progesterone (P₄) prevented the termination of pregnancy in Ovx animals, but 1 mg did not. Treatment with estradiol 17β (E₂) in addition to 4 mg of P₄ tended to result in a lower rate of fetal survival than that of the Ovx group treated with 4 mg of P₄ alone. With regard to fetal weight, treatment with 4 mg of P₄ resulted in the same weight as in the sham-operated controls, but the addition of 0.2 or 1 μg of E₂ to the 4 mg of P₄ resulted in a significantly lower weight than that of fetuses in the 4 mg of P₄ group. The present study suggests that adequate maintenance of pregnancy in ovariectomized gerbils can be achieved by daily treatment with 4 mg of P₄ alone. Moreover, treatment with 0.2 or 1 μg of E₂ in addition to 4 mg of P₄ caused a deterioration in the maintenance of gestation, in contrast to the effects in rats, mice and hamsters.