  A transformed fish cell line expressing a green fluorescent protein-luciferase fusion gene responding to cellular stress; Carpeaux, Rudy  ; Martial, Joseph et alin Toxicology in Vitro (2002), 16(2), 201-7 We obtained a stable transformed fish (EPC) cell line containing a reporter gene under the control of the tilapia HSP70 promoter. Expression of the reporter gene, coding for a green fluorescent protein ... [more ▼] We obtained a stable transformed fish (EPC) cell line containing a reporter gene under the control of the tilapia HSP70 promoter. Expression of the reporter gene, coding for a green fluorescent protein (GFP)-luciferase fusion protein, was assessed by measuring the luciferase enzymatic activity by luminometry and the GFP expression by fluorescence microscopy and flow cytometry. The clone was characterized for its capacity to respond to heat shock treatment. The results show high induction after 1 h at 37 degrees C of treatment, up to 500-fold. In addition, its convenience to detect a large range of cellular stressors was evaluated. We observed high induction when Cd2+, Zn2+, Hg2+ or Cu2+ was added, but not Pb2+. In addition, activation of the reporter gene was observed in the presence of other compounds such as acetyl chloride, tetrachlorophenol, chloroacetamide and sodium arsenite. In conclusion, this cell line can be used as a rapid, cheap and easy biological test to determine cellular stress induced by environmental pollutants, alone or in conjunction with other, more specific assays. [less ▲] Detailed reference viewed: 43 (5 ULg)In vitro ototoxicity of aminoglycosides and platin derivatives. A semi-automatic assay for sensory hair cell damage in explanted rat organ of corti.; LEFEBVRE, Philippe ; et alin Toxicology in Vitro (1998) The ototoxic damage that drugs such as neomycin, kanamycin, colistin, cisplatin, transplatin and carboplatin cause on outer and inner hair cells in postnatal day 3 rat cochlear explants was investigated ... [more ▼] The ototoxic damage that drugs such as neomycin, kanamycin, colistin, cisplatin, transplatin and carboplatin cause on outer and inner hair cells in postnatal day 3 rat cochlear explants was investigated. Phalloidin-fluorescein conjugate-stained stereocilia bundles of sensory hair cells were quantified by video image analysis as a measurement of ototoxic effect. The video image quantification system established dose-response curves for ototoxic drugs (e.g. calculation of an IC50) and allowed comparisons between several ototoxins from the same family. This methodology provided the means to assess the efficacy of otoprotectant agents in preventing ototoxicity. Poly-l-aspartate (10-5M) and poly-l-glutamate (10-5M) protected auditory hair cells from neomycin (10-3M) toxicity while reduced glutathione (10-3M) provided protection against cisplatin (10-4M)-induced hair cell damage. [less ▲] Detailed reference viewed: 10 (0 ULg) |
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