RESUMO
The effects of limb blastemas of Pleurodeles waltl on axon growth from fragments of spinal cord were studied in vitro. Cultured in a defined medium, spinal cord fragments regenerated sparse, short axons. The culture of spinal fragments in the presence of blastemas greatly enhanced the length, number and survival of axons. Testing separately each of the two components of the blastema showed that only the mesenchyme exerts a neurotropic effect on the spinal fragments. Other tissues such as muscle or skin had a limited neurotrophic effect. Additionally, the neurotrophic activity of blastemas seems to be dependent of its proliferation status. Compared with blastemas of regenerating limbs from young animals, irradiated blastemas (devoid of mitotic activity) and blastemas of regenerating limbs from old animals or differentiated blastemas (both characterized by a low mitotic activity), exhibited a weaker neurotrophic influence. The blastema neurotrophic factor is not an attachment molecule but a soluble one and cannot be nerve growth factor (NGF) or fibroblast growth factor (FGF). It has a relatively low molecular weight (less than 15 kDa) and its protein nature was ascertained by its sensitivity to heating and proteases. As the production of this mesenchyme-derived neurotrophic factor depends upon mesenchymal cell proliferation of the blastema, we suggest that there is loop of positive regulation between spinal nerves and blastema. Blastema tissues may stimulate nerve regeneration allowing the stimulation of proliferation of blastema cells by regenerating nerve fibers. Alternatively, blastema cells may produce a neurotrophic factor whose secretion might be dependent on cell proliferation.
Assuntos
Cotos de Amputação/inervação , Axônios/fisiologia , Botões de Extremidades/inervação , Regeneração Nervosa/fisiologia , Pleurodeles/fisiologia , Medula Espinal/citologia , Animais , Contagem de Células/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Meios de Cultivo Condicionados/química , Meios de Cultivo Condicionados/farmacologia , Técnicas de Cultura , Membro Anterior/inervação , Botões de Extremidades/citologia , Botões de Extremidades/crescimento & desenvolvimento , Botões de Extremidades/efeitos da radiação , Mesoderma/citologia , Peso Molecular , Fatores de Crescimento Neural/biossíntese , Fatores de Crescimento Neural/química , Fatores de Crescimento Neural/farmacologia , Regeneração Nervosa/efeitos dos fármacos , Medula Espinal/efeitos dos fármacos , Medula Espinal/fisiologiaRESUMO
Rabies infection of domestic and wild animals is a serious problem throughout the world. The major disease vector in Europe is the red fox (Vulpes vulpes) and rabies control has focused on vaccinating and/or culling foxes. Culling has not been effective, and the distribution of five vaccine baits is the only appropriate method for the vaccination of wild foxes. Although some European countries have conducted field vaccination campaigns using attenuated rabies virus strains, their use has not been extensively approved because they retain pathogenicity for rodents and can revert to virulence. These strains cannot be used in North America because they are pathogenic for the striped skunk (Mephitis mephitis) and are ineffective in the raccoon (Procyon lotor). We have constructed a recombinant vaccinia virus, VVTGgRAB, expressing the surface glycoprotein (G) of rabies virus (ERA strain). The recombinant was a highly effective vaccine in experimental animals, in captive foxes and in raccoons. We report here the results of a large-scale campaign of fox vaccination in a 2,200 km2 region of southern Belgium, an area in which rabies is prevalent. After distribution, 81% of foxes inspected were positive for tetracycline, a biomarker included in the vaccine bait and, other than one rabid fox detected close to the periphery of the treated area, no case of rabies, either in foxes or in domestic livestock, has been reported in the area.
Assuntos
Raposas , Vacina Antirrábica/administração & dosagem , Raiva/prevenção & controle , Vacinas Sintéticas/administração & dosagem , Animais , Bélgica/epidemiologia , Europa (Continente)/epidemiologia , Imunização , Incidência , Raiva/epidemiologia , Vacina Antirrábica/uso terapêutico , Vírus da Raiva/genética , Vírus da Raiva/imunologia , Guaxinins , Especificidade da Espécie , Vacinas Sintéticas/uso terapêutico , Vaccinia virus/genética , Células VeroRESUMO
The vaccination of wild animals against rabies has been developed most extensively in Europe. Experiments have demonstrated the efficacy of a vaccinia-rabies recombinant virus administered by the oral route in foxes. The innocuity of this vaccine was tested in the target species as well as in several non-target wild and domestic species. Because of its safety and heat-stability, this recombinant virus should offer an excellent alternative to the attenuated strains of rabies virus currently used in the field. A large scale field trial was conducted in Belgium in October 1988 to assess the efficacy of this new vaccine-bait systems.
Assuntos
Raposas , Vacina Antirrábica/administração & dosagem , Raiva/veterinária , Vacinação/veterinária , Vacinas Sintéticas/imunologia , Vacinas/imunologia , Vacínia/imunologia , Vacinas Virais/imunologia , Animais , Gatos , Muridae/imunologia , Raiva/prevenção & controle , Vacina Antirrábica/efeitos adversos , Vírus da Raiva/imunologia , Temperatura , Vacínia/genéticaRESUMO
A field trial of fox vaccination against rabies using a vaccinia-rabies recombinant virus was carried out in Belgium on October 24, 1987. Each vaccine capsule contained a suspension of 10(8) TCID50 of the recombinant virus and was introduced into a chicken head. Each chicken head contained 150 mg of tetracycline as a marker of uptake. Two hundred and fifty heads were distributed in an area of 6 km2 situated within a military zone. The bait uptake was monitored for 15 days after the distribution. Sixty-three per cent of the chicken heads were taken by wild animals within that period. The trial was controlled according to the rules defined by the World Health Organisation.
Assuntos
Raposas , Vacina Antirrábica , Raiva/veterinária , Vacinação/veterinária , Vacinas Virais , Animais , Bélgica , Humanos , Raiva/prevenção & controle , Vacina Antirrábica/administração & dosagem , Tetraciclina/análise , Fatores de Tempo , Vacinas Sintéticas/administração & dosagem , Vaccinia virus , Vacinas Virais/administração & dosagemRESUMO
In order to approach the problem of regulation of growth factor(s) production during limb regeneration in newt, we co-cultivated spinal cord segments and blastemas. First we showed that, like the sensory supply, the spinal cord possesses size-dependent mitogenic capacities for limb blastemas. A 5-mm long spinal segment enhances radiolabelled thymidine incorporation to the same extent as spinal ganglia (1.6-fold). Second, we co-cultivated blastemas with spinal segments, the nerve fibers of which were previously stimulated to regenerate (= stimulated spinal segment) or not (= non-stimulated spinal segment). Only after a 24-h coculture, do stimulated spinal segments enhance thymidine incorporation in blastemas 2-fold more than non-stimulated spinal segments. Our results suggest that during limb regeneration brachial nerves produce more growth factor(s) when regrowing, inducing the proliferation of blastema cells which in return deliver a neuronotrophic factor acting on these nerves.