Localization of extracellular matrix receptors in ICGN mice, a strain of mice with hereditary nephrotic syndrome.

Fibrotic degeneration was examined in the kidneys of ICR-derived glomerulonephritis (ICGN) mice, a novel inbred mouse line with a hereditary nephrotic syndrome of unknown etiology considered to be a good model of human idiopathic nephrotic syndrome. In the present study, we histochemically revealed changes in accumulation of extracellular matrix (ECM) components and in localization of integrins, cellular receptors for ECM, in the kidneys of ICGN mice with the progression of renal failure. Excessive accumulation of basement membrane (laminin and collagen IV) and interstitial (type III collagen) ECM components were demonstrated in the glomeruli and tubulointerstitum of ICGN mice. Marked deposition of type I collagen and tenascin was seen only in the glomeruli of ICGN mice but not in those of ICR mice as normal controls. Increased expression of integrin alpha1-, alpha2-, alpha5- and beta1-subunits in glomeruli with fibrotic degeneration and abnormal distribution of alpha6-subunit were noted in the kidneys of ICGN mice. Excessive laminin, a ligand of alpha6beta1-integrin, was demonstrated on the tubular basement membrane, but alpha6-subunit diffusely disappeared on the basal side of the tubular epithelial cells. We presumed that abnormal integrin expression in renal tubules causes epithelial cell detachment, and consequently tubular nephropathy, and results in disorder of ECM metabolism causing excessive accumulation of ECM components in the kidneys of ICGN mice.     

Spontaneous regression of bovine cutaneous leukosis

Biopsies of skin from a 2-year-old heifer with spontaneously regressing dermal leukosis were examined. The heifer was not infected with bovine leukemia virus and was negative for tumor-associated antigens of enzootic bovine lymphosarcoma. By hematological standards for bovine leukemia, the heifer was positive at about 60 days post-occurrence of the disease (POD). At 53 days POD, lymphoblastic neoplastic cells in the dermis reacted with anti-T lymphocyte monoclonal antibody by the avidin biotin peroxidase complex method. The cells had intracytoplasmic clustered dense bodies under electron microscopy. From 53 to 83 days POD, figures of the transepithelial elimination (TE) against neoplastic cells and perivascular infiltration of small lymphocytes were in the dermis. Small lymphocytes reacted with anti-T lymphocyte monoclonal antibody. At necropsy there were no neoplastic lesions; there were flat lymph node-like tissues in the subcutis. Many germinal centers were seen in the lymphatic organs. Blood lymphocytes at 46 days POD were stimulated by phytohemagglutinin-P and concanavalin A. Sera, taken until 75 days POD and at necropsy, showed an inhibitory effect on mitogen-induced blastogenesis of normal bovine lymphocytes. These results suggested the existence of a spontaneous regressive mechanism against neoplastic lesions by TE and tumor immunity.     

Sequence of the canine c-yes proto-oncogene

Cloning of the canine yes oncogene was attempted from a c library derived from a healthy canine spleen using a human c-yes-1 probe. The nucleotide and amino acid sequences revealed that the canine yes gene contained an open reading frame consisting of 539 amino acids. Its product had a molecular mass of 60,368 Daltons and showed 95·9 per cent and 90·4 per cent homology with human and chick p61^c-yes, respectively. Moreover, the product had a myristylation signal, src homology region (SH) 3, SH2, and tyrosine kinase domains showing 98·8 per cent and 96·0 per cent homology with those of human beings and chickens, respectively. These findings indicate that the products of the canine yes gene may have non-receptor-type tyrosine kinase activity on the cell membrane, as is the case in human and chick p61^c-yes     

Monitoring of genetic diversity in the Japanese Black cattle population by the use of pedigree information

The gene pool of the Japanese Black cattle has been completely closed to foreign breeds during the last 100 years. Genetic diversity of the Japanese Black cattle from 1960 to 2000 was monitored with three estimates of effective number of ancestors. Founder genome equivalent (N_ge) accounts for all the causes of reduction of diversity. Effective number of founders (N_ef) and non‐founders (N_enf) explain reduced diversity because of unequal genetic contributions of founders and random genetic drift in non‐founders, respectively. Further examination using gene dropping simulation was conducted to obtain information on survival of founder alleles. Unique founder alleles were dropped down along the actual pedigree with Monte Carlo procedure following Mendelian segregation rules, and generated genotypes of all the current live animals (612 959 heads). Pedigree records consisted of 2 075 188 animals was used for these analysis. The estimates of three effective numbers (N_ef, N_ge, and N_enf) decreased from 418.6 to 50.3, 86.6 to 7.3, and 109.2 to 8.5, respectively, during the period 1960–2000. The increasing differences between two kinds of genetic diversity indices derived from N_ge and N_ef showed that large part of the reduced diversity from 1980 was attributed to genetic drift caused by the intensive use of particular limited number of sires. In gene dropping analysis, probabilities of extinction of founder alleles were derived from their distributions of frequency in the current animals. Several founders showed low probabilities of allele extinction, irrespective of their relatively low genetic contributions. This suggests that these founders have lineages through which their alleles are surely transmitted to the current breed. The use of these founders as a strategy for recovering the genetic diversity was discussed.     

Histological and immunohistochemical evaluation of granulosa cells during different stages of folliculogenesis in bovine ovaries

Bovine granulosa cells (GC) vary in their morphological aspect during different stages of folliculogenesis. In this study, 10 morphologically normal bovine ovaries were collected to study the structural aspects of different stages of GC using intermediate filament protein antibodies including cytokeratin AE1/AE3 (AE1/AE3), vimentin, nectin‐4 and desmin. Hormonal immunolocalization was assessed using the immunomarkers anti‐Müllerian hormone (AMH) and inhibin alpha. In addition, tumour markers and proliferation markers using c‐erbB‐2 oncoprotein and proliferating cell nuclear antigen, respectively, were investigated. The immunolabelling of AE1/AE3 in GC was strongest in the early follicle stage and gradually decreased when reaching the Graafian follicle stage. Its immunolabelling increased again as the stage progressed from stage I to stage III. The immunolabelling of inhibin alpha was inversely proportional to that of AE1/AE3 in the developing ovarian follicles as their immunolabelling is opposite to each other during folliculogenesis. AMH was immunopositive in almost all GC stages in different intensities and percentages, except for some negative staining in the atretic IV follicles. The atretic IV follicle is a unique type of atretic follicle that shows Call‐Exner body formation, which was mainly found in older cows in this study. The distinct patterns of immunoreactivity for various types of immunomarkers in the different GC stages will play an important role in diagnostic assistance of various follicle conditions, including cystic ovaries and GC tumours.     

Nitrogen fertilization affects yields and storage compound contents in seeds of field-grown soybeans cv Enrei (Glycine max. L) and its super-nodulating mutant En-b0-1 through changing N2 fixation activity of the plants

ABSTRACT

We compared the protein and oil contents, seed yields, and natural abundance of ¹⁵N (σ¹⁵N) of seeds from the plants of the cultivar Enrei, which has moderate nodulating ability (Enrei) with those of its two near-isogenic lines (NILs), a super-nodulating genotype of Enrei, i.e., En-b0-1, and a non-nodulating genotype of Enrei, i.e., En1282. Plants of these three genotypes were grown on four types of field plots with different types of urea coated slow-release nitrogen (N) fertilizers (CUSLNFs) which delivered N in different manners during plant growth . The seed yield of the En1282 plants was similar to that of the Enrei plants but much greater than that of the En-b0-1 plants when the plants were grown on the field to which a CUSLNF with a long lifespan was applied. The amounts of protein per seed were positively correlated with those of oil per seed in the case of En1282 plants irrespective of the field N conditions. The seed protein contents were proportional to the seed weight in both En1282 and Enrei plants. Such a relationship was not observed in the En-b0-1 plants, and the protein and oil contents in the seeds of En-b0-1 plants were negatively correlated with each other. These observations suggested that the N supply to maturing seeds was a key to the protein contents in the En1282 and Enrei plants and that the C supply to maturing seeds was a key to the protein contents in the En-b0-1 plants. The comparison of the σ¹⁵N values, protein contents, and seed yields of seeds from En1282 plants suggested that the Enrei plants assimilated considerable amounts of N from the soil during the late plant growth. We concluded that high N₂ fixation activity depressed the amounts of oil per seed and changed the protein and oil contents in soybean seeds.     

Molecular characterization of the equine herpesvirus 1 strains RacL11 and Kentucky D

The pathogenicities of RacL11 and Kentucky D strains of equine herpesvirus 1 in the hamster infection model are different from those of Ab4p and the Japanese isolates. Virus genome restriction fragment length polymorphism analysis and sequence comparison of an intergenic region, glycoproteins and tegument genes showed higher conservation but with some strain-specific differences. These results indicate that point nucleotide differences in RacL11 and Kentucky D might be responsible for their pathogenicity in rodent models.     

Diffuse bilateral hemangiosarcoma of the uterus in a dog

A 15-year-old female mongrel dog showed abdominal swelling, marked hemorrhagic ascites and vulvar discharge, and ovariohysterectomy was performed. Grossly, the uterus was enlarged bilaterally without apparent mass formation. Histologically, the uterine muscular wall was composed of proliferated sinusoidal vessels. In some areas, irregular and small vessels proliferated markedly, while in others, pleomorphic and atypical tumor cells forming irregular vascular structures were predominant. From these findings, the case was diagnosed as diffuse bilateral hemangiosarcoma of the uterus that invaded to the ovary and broad ligament. The relationship between the tumor and angiomatosis was discussed.