RESUMEN
The zebrafish has emerged as a powerful genetic model of cancer, but has been limited by the use of stable transgenic approaches to induce disease. Here, a co-injection strategy is described that capitalizes on both the numbers of embryos that can be microinjected and the ability of transgenes to segregate together and exert synergistic effects in forming tumors. Using this mosaic transgenic approach, gene pathways involved in tumor initiation and radiation sensitivity have been identified.
Asunto(s)
Transformación Celular Neoplásica/genética , Transformación Celular Neoplásica/efectos de la radiación , Técnicas de Transferencia de Gen , Microinyecciones/métodos , Neoplasias Inducidas por Radiación/genética , Animales , Animales Modificados Genéticamente , Fase de Segmentación del Huevo , Proteínas de Unión al ADN/administración & dosificación , Proteínas de Unión al ADN/genética , Embrión no Mamífero , Genes bcl-2 , Genes myc , Genes p53 , Proteínas Fluorescentes Verdes/administración & dosificación , Proteínas Fluorescentes Verdes/genética , Proteínas Luminiscentes/administración & dosificación , Proteínas Luminiscentes/genética , Proteínas Mutantes/genética , Proteínas Nucleares/administración & dosificación , Proteínas Nucleares/genética , Leucemia-Linfoma Linfoblástico de Células T Precursoras/genética , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas p21(ras) , Transgenes , Pez Cebra/embriología , Proteínas ras/genética , Proteína Fluorescente RojaRESUMEN
A novel, membrane potential sensitive dye and a fluorescence imaging plate reader (FLIPR) have been used to characterize the pharmacological properties of rat Na(v)1.8 voltage-gated sodium channels (VGSC) in parallel with rat Na(v)1.2a and human Na(v)1.5 VGSC subtypes, respectively. The sensitivity of recombinant Na(v)1.2a-CHO, Na(v)1.5-293-EBNA, and Na(v)1.8-F-11 cells to VGSC activators was subtype dependent. Veratridine evoked depolarization of Na(v)1.2a-CHO and Na(v)1.5-293-EBNA cells with pEC(50) values of 4.78 +/- 0.13 and 4.84 +/- 0.12, respectively (n = 3), but had negligible effect on Na(v)1.8-F-11 cells (pEC(50) < 4.5). Type I pyrethroids were without significant effect at all subtypes. In contrast, the type II pyrethroids deltamethrin and fenvalerate evoked direct depolarization of Na(v)1.8-F-11 and Na(v)1.5-293-EBNA cells. Deltamethrin potentiated the veratridine-evoked response in Na(v)1.8-F-11 cells by > or =20-fold, in contrast to a Asunto(s)
Proteínas Musculares/metabolismo
, Proteínas del Tejido Nervioso/metabolismo
, Canales de Sodio/metabolismo
, Sodio/metabolismo
, Animales
, Colorantes Fluorescentes
, Humanos
, Canal de Sodio Activado por Voltaje NAV1.5
, Canal de Sodio Activado por Voltaje NAV1.8
, Técnicas de Placa-Clamp
, Ratas
, Factores de Tiempo