ABSTRACT
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.
Subject(s)
Cell Transformation, Neoplastic/genetics , Cell Transformation, Neoplastic/radiation effects , Gene Transfer Techniques , Microinjections/methods , Neoplasms, Radiation-Induced/genetics , Animals , Animals, Genetically Modified , Cleavage Stage, Ovum , DNA-Binding Proteins/administration & dosage , DNA-Binding Proteins/genetics , Embryo, Nonmammalian , Genes, bcl-2 , Genes, myc , Genes, p53 , Green Fluorescent Proteins/administration & dosage , Green Fluorescent Proteins/genetics , Luminescent Proteins/administration & dosage , Luminescent Proteins/genetics , Mutant Proteins/genetics , Nuclear Proteins/administration & dosage , Nuclear Proteins/genetics , Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/genetics , Proto-Oncogene Proteins/genetics , Proto-Oncogene Proteins p21(ras) , Transgenes , Zebrafish/embryology , ras Proteins/genetics , Red Fluorescent ProteinABSTRACT
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 Subject(s)
Muscle Proteins/metabolism
, Nerve Tissue Proteins/metabolism
, Sodium Channels/metabolism
, Sodium/metabolism
, Animals
, Fluorescent Dyes
, Humans
, NAV1.5 Voltage-Gated Sodium Channel
, NAV1.8 Voltage-Gated Sodium Channel
, Patch-Clamp Techniques
, Rats
, Time Factors
