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Combinatory Chemical and Biological Approaches to Investigate Metal Elements in Agricultural Runoff Water

^a Centre de Biogéographie-Ecologie, FRE 2545 CNRS/ENS-LSH, 15 Parvis René Descartes, 69366 Lyon cedex 07, France
^b INRS–Eau, Terre & Environnement, Université du Québec, 2800 rue Einstein, C.P. 7500, Sainte-Foy (Québec), G1V 4C7, Canada
^c Vigicell, Institut André Lwoff, Bât. G, 7 rue Guy Moquet, 94801 Villejuif, France

View larger version (32K): [in a new window]   Fig. 1. Metallothionein-2A (MT-2A) gene expression by CEM-C12 cells exposed to water samples. Cells were incubated (1.5 x 106 cells mL–1) for (A) 6 and (B) 18 h in medium reconstituted with either water samples (1–10) or ultrapure water as control (C) as described in the Materials and Methods section. Cadmium chloride (5 µM) was used as a positive control (Cd). Total cellular RNA was extracted and subjected to reverse transcriptase–polymerase chain reaction (RT–PCR) analysis. As described in the Material and Methods section, [-32P]-dCTP was added to the PCR mixture and quantification of PCR products was done by measuring the amount of 32P-radiolabeled nucleotide (cpm) incorporated into amplified fragments. The 32P-labeled bands of PCR products corresponding to MT-2A and glyceraldehyde-3'-phosphate dehydrogenase (GaPDH; housekeeping gene) are presented. (C) For 18 h of exposure, MT-2A gene expression was evaluated by the signal ratio of cpm values for MT-2A on respective cpm values for GaPDH. The RT–PCR analysis was performed in triplicate (standard deviation was always less than 15% of the mean). Results are expressed as fold of induction over control (signal ratio from cells cultured with water sample to signal ratio from cells cultured with ultrapure water).

View larger version (15K): [in a new window]   Fig. 2. Representation of metallothionein-2A (MT-2A) gene expression by CEM-C12 (as pictured in Fig. 1C) versus Ca concentration measured in water samples. Numbers close to each dot represent number of water samples. Sample 7, which is considered as an outlier in this series of points, is represented by an empty dot.