["A man is being made"].
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Biological Evolution↗
Biomedical subjects
M Wyder
Publications and source records attributed to M Wyder.
Canine distemper virus associated proliferation of canine footpad keratinocytes in vitro.
Infection of canine footpads with canine distemper virus (CDV) can result in so-called hard pad disease characterized by footpad epidermal proliferation and hyperkeratosis. Cultured canine footpad keratinocytes (CFK) were inoculated with a virulent canine distemper virus strain (A75/17-CDV) to study the effects of CDV-infection on keratinocyte proliferation. Infection was analyzed by immunohistochemistry and in situ hybridization for CDV nucleoprotein (N-protein) antigen and mRNA. CDV caused a persistent, non-cytocidal infection with spread from single cells to infection of the confluent cell layer 7 days post infection (p.i.). Absolute cell numbers were significantly higher in infected cultures compared to control cultures from day 4 until day 6 p.i. Infected cultures contained significantly more total DNA on day 5 p.i. compared to controls. Immunohistochemical investigation of proliferation markers Ki67 and BrdU demonstrated a nearly two-fold increase in numbers of positive cells on day 5 p.i. compared to controls. These findings demonstrate that canine distemper virus infection of canine footpad keratinocytes in vitro was associated with proliferation.
Animals↗
Cultivation and characterisation of primary and subcultured equine keratinocytes.
We describe the establishment and characterisation of equine keratinocyte cultures with maintenance of a high proliferative capacity up to the second passage. Improved attachment and growth were obtained by seeding primary cells on equine feeder layers. Subcultured keratinocytes showed optimal growth when seeded on collagen type I. The proliferation rate of cells on this substrate exceeded that seen for cells seeded on equine feeder layers. By immunohistochemistry, epithelial origin and state of differentiation of the equine keratinocytes were determined. They expressed keratin and desmoplakin I/II, but lacked keratin 10. Electron microscopy revealed typical features of cultured keratinocytes. Purity of keratinocyte cultures was determined by vimentin staining. This is the first report on the establishment of equine keratinocytes derived from lip epithelium. It forms the basis to study equine keratinocyte biology and the pathogenesis of epidermal diseases. Since wound healing represents a severe problem in equine dermatology, our data may be essential for the establishment of new and improved therapy.
Animals↗
Cloning of canine Dsg1 and evidence for alternative polyadenylation.
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Alternative Splicing↗
Periplakin and envoplakin are target antigens in canine and human paraneoplastic pemphigus.
BACKGROUND: On the basis of clinical and histopathologic similarities to human paraneoplastic pemphigus (PNP), we recently identified the first case of PNP in a nonhuman species, the dog. OBJECTIVE: To determine a similar pathogenesis in both species, the present study aimed to define whether common antigens are targeted in dog and man. METHODS: Canine and human PNP sera were used in parallel to immunoprecipitate 14C-labeled human keratinocyte antigens. The immunoreactive proteins were then identified by immunoprecipitation of canine keratinocyte extracts with specific antibodies to the antiplakin family members follwed by immunoblot analysis using canine and human PNP sera. RESULTS: Protein bands of 210, 190, 170, and 130 kd were identified in dogs and humans. In both species, envoplakin and periplakin were demonstrated as antigens. Anti-desmoglein 3 antibodies could not be demonstrated in canine PNP, but in human PNP. CONCLUSION: These results demonstrate that canine PNP closely correlates to the human counterpart and may therefore represent an excellent model for the human disease.
Animals↗
Behavioral model of chronic tinnitus in rats.
An animal model of tinnitus was developed to study chronic salicylate-induced tinnitus in rats. Novel features of the model included oral dosing of salicylate, test stimuli that included a range of pure tones and silence, and assessment of tinnitus for several months. Experimental subjects were given sodium salicylate in their drinking water while control subjects received normal tap water. Subjects were conditioned to press a lever for food in the presence of continuous white noise. At random intervals, offset of the noise was paired with a noxious stimulus, resulting in cessation of lever pressing during the silent test periods. At other randomly scheduled intervals, a test tone was substituted for the white noise, unpaired with noxious stimuli. When the test stimuli were pure tones, the salicylate-treated subjects suppressed less than the control subjects. One explanation for this result is that the experimental subjects' sensations of tones were noisier than those of the controls because experimental subjects were experiencing tinnitus.
Acoustic Stimulation↗
Loss of invasiveness in squamous cell carcinoma cells overexpressing desmosomal cadherins.
The molecular and structural characteristics of intercellular adhesion were investigated in a squamous cell carcinoma (SCCA) cell line, originally derived from an oral tumor with an invasive growth pattern. The expression of adherens junction and desmosomal components were compared with that of cultured normal oral keratinocytes. Lack of membrane association in interdesmosomal areas, the disorganization of the actin cytoskeleton and the faster cell disassembly upon E-cadherin antibody binding in SCCA cells indicated decreased functional adherens junctions. These observations were supported by a significant reduction in E-, N-, and P-cadherin protein expression. In contrast, the level of desmosomal cadherin proteins, desmoglein 1/2 and desmocollin 2, were substantially upregulated and accompanied, ultrastructurally, by increased number and size of desmosomes. Since tumor invasion suppressor capacity has been proposed for desmosomal cadherins, we investigated the in vivo invasion potential of these SCCA cells by introducing them into SCID mice. Tumors developed, but with a benign phenotype. Based on these results, we hypothesize that the benign behavior of this SCCA cell line is a consequence of overexpressed desmosomal cadherins. This SCCA cell line, therefore, represents an excellent model system to further investigate the regulation and tumor invasion suppressor potential of desmosomal adhesion molecules.
Animals↗