Biomedical subjects
Publications and source records attributed to W Schleger.
Short term cultures of four feline fibrosarcomas were analysed cytogenetically. There was marked genetic heterogeneity between the four cats, each showing a different clonal abnormality. The aberrations detected were one deleted B2, one marker F1 and two reciprocal translocations, t (A2q; E3q) and t (A1q; B4p).
Four solid tumours (one haemangiopericytoma, one haemangioendothelioma, one spindle-cell sarcoma and one mammary carcinoma) in dogs were analysed cytogenetically. In the haemangiopericytoma, an additional small chromosomal segment was present. Very complex changes including centric fusions and symmetric meta-centrics 1, 6, 10 and 12 were conspicuous in the highly unbalanced karyotype of the haemangioendothelioma. Complex changes, particularly many centric fusions and a tandem translocation 4/14, were features of the spindle-cell sarcoma. One centric fusion and a symmetric metacentric 13 were present in the mammary carcinoma.
An 11-year-old female cat and a 12-year-old male cat developed fibrosarcomas. Trisomy D1 was observed in a high percentage (20%) of the tumour cells of the female patient while a large marker chromosome F1 was present in 25% of the tumour cells of the male patient.
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In a 13-year-old, male rough-haired Dachshund with a malignant melanoma, cytogenetic evaluation of tumour cells showed hyperdiploidy (79 to 81 chromosomes) in 50 per cent of the metaphases. Several bi-armed chromosomes (centric fusions, one isochromosome and two unidentified markers) were observed.
Cells of mammary sarcomas in three dogs were analysed cytogenetically. In an osteoidsarcoma, hyperdiploidy with a range of 92 to 98 chromosomes, and several structural aberrations (for example, a derivative chromosome 1, isochromosome 13 and several bi-armed markers) were observed. Isochromosome 13 was also present in a case of an osteoidchondrosarcoma. In a case of chondro-osteosarcoma both hypodiploidy with a chromosome range of 60 to 66 and hyperdiploidy with a range of 115 to 128 and several centric fusions were observed.
The permanent bovine cell line, Madin-Darby bovine kidney, was cytogenetically analyzed. Chromosomal staining indicated 52 metaphase chromosomes with variations from 49 to 54. The isochromosomes 5, 12, and 13, and centric fusions between chromosomes 1 and 26, 9 and 11, 9 and 24, 17 and 25, 18 and 23, 20 and 24, and 24 and 27 were considered marker chromosomes.
An eight-year-old male cat developed a sarcoma. The cytogenetic evaluation of the tumour cells showed the presence of hyperdiploidy (range 40 to 46 chromosomes). This hyperdiploidy was encountered in all the cells examined. Extra-chromosomes numbers C1, C2, B4, D4 and E3 were mainly responsible for the hyperdiploid chromosomal complements. There was a high incidence of monosomy E3.
Two dogs developed osteosarcomas. In both cases, the cytogenetic analysis revealed the presence of numerically and structurally altered karyotypes. The chromosome ranges were 90 to 99 and 65 to 67, respectively. Centric fusions frequently identified were 1/3, 1/30 and 2/9 in one case, 3/19, 12/18 and 13/17 in the other.
A 14-year-old German shepherd dog developed an alveolar adenoma of the thyroid. The cytogenetic evaluation of the tumour cells showed two ranges of chromosome numbers. Centric fusions 2/32, 5/26, 6/11, 8/11, 9/23 and 14/31 were G-band identified in this complex change of karyotype.
Tumor cells of two cases of mammary neoplasms in dog were investigated cytogenetically. An 11-year-old cocker spaniel showed a marked X-chromosome in most metaphases (80%). In a 12-year-old poodle, tumor cells with the following abnormalities were found: trisomy 1, monosomy X and two marker chromosomes.
In a 9-year-old female dog (Basset Artesian Normand) with a mammary adenocarcinoma, cytogenetic evaluation of tumor cells showed a chromosome number of 76 in most metaphases (95%). The following abnormalities were found: symmetric metacentric chromosomes 1 and 6, centric fusion 3/38, a marker X-chromosome (Xmar) and a biarmed small marker chromosome (mar). In the remaining metaphases (5%) there were additional biarmed marker chromosomes present.
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Bovine follicular oocytes were matured in TCM 199 supplemented with: (1) fetal calf serum (FCS, 20% v/v), Luteinizing Hormone (LH, 10 ug/ml), and Estradiol-17-beta (E(2), 1 ug/ml) in Experiment 1; (2) 20% cow serum recovered at standing estrus (Experiment 2); or (3) 20% FCS (Experiment 3). Maturation, fertilization, and initial cleavage development were evaluated at 16 and 48 h after in vitro insemination. The proportions of oocytes fertilized after maturation in the presence of added hormones (78.5%, Experiment 1) or estrous serum (71.3%, Experiment 2) were significantly higher (p < 0.01) than after use of FCS alone (39.3%, Experiment 3). Cleavage of zygotes within 48 h post-insemination differed significantly (p < 0.01) between maturation treatments, 27.3%, 75.5% and 6.6% for Experiments 1, 2, and 3, respectively. Results demonstrate a beneficial influence of estrous cow serum, characterized by an elevated concentration of LH, on bovine oocyte maturation in vitro.
An eight-year-old female cat developed a skin neurofibroma. The cytogenetic evaluation of the tumour cells showed the presence of a high percentage (16.4 per cent) of trisomic cells. The trisomy concerned chromosome number D2.
Chromatin configurations of cattle oocytes from different follicle sizes were classified in three chromatin categories. Early diplotenes, dictyate-like structures, more progressed diplotenes and early diakinesis and metaphases were found. In follicle group A (0.4-0.9 mm) immature diplotenes predominated (82.3%), and a rather low maturation competence was observed (12.1%). With increasing follicle size the maturation competence was improved to 47.8% metaphases II in follicle group C (3-8 mm). Impressive CMA3 positive blocks were visualized especially in early diplotenes. Heterochromatic CMA3 positive clusters surrounded the nucleolus.
The expression of nucleolus organizer region (NOR) activity in diploid cells was investigated in a model mammalian hybrid system, the hinny (female ass x male horse), by sequential Ag-NOR and chromomycin A3/distamycin A/DAPI (CDD) staining ion lectin-stimulated peripheral blood lymphocytes. In the majority of cases we found non-expression of the horse-derived NOR chromosomes in the hinny. However, in one case there was strong NOR expression on horse-derived chromosome no. 1.
Cytogenetic studies of the mithun (Bos frontalis), the siri (Bos indicus) and their hybrids, as well as histological examinations of testes and epididymis of siri and hybrids were undertaken to throw further light on male hybrid infertility and interspecies incompatibility, in order to facilitate the possible development of a stable interspecies crossbreed which would be similar to the highly profitable jatsum, the female mithun cross siri hybrid. In both species there are distinct centromeric heteromorphisms which should provide a rich source of genetic markers for tracing chromosomes in controlled hybrid programmes. The nucleolar organisers in the F1 hybrids were derived from both parents, in contrast to other interspecies hybrids where the nucleolar organisers of one species tend to be suppressed by the other. This indicates a relatively close relationship between mithun and siri and supports the prospect of success for developing a stable crossbreed. Transferrins which may also be a useful source of genetic markers were identified and differences between siris and mithuns noted, especially in the D-bands. No A-bands were found in the siris and no E-bands in the mithuns. Whether this is a breed or species characteristic will have to be confirmed by examination of more animals. The findings on blood groups and haemoglobins, including differences between the two species, were similar to results obtained previously. In meiosis the sex vesicles were intact which indicates that infertility and incompatibility factors should be located in autosomes. In cytogenetic and histological examinations the testes of the siris were comparable to those of European cattle, while in all hybrids spermatogenetic activity was deficient although there were great differences between and within generations. While the exact mechanism of this diversity is not known at this stage, it should make it possible to select for increased male hybrid fertility which would be a prerequisite for developing a stable interspecies crossbreed. This selection would be facilitated by correlating infertility and incompatibility with genetic profiles and markers. A mithun cross siri crossbreed would also serve as a model for other interspecies breeding.