Metallothionein induction in the RTH-149 cell line as an indicator for heavy metal bioavailability in a brackish environment: assessment by RT-competitive PCR.
Estuarine and marine near-shore environments are often subjected to heavy metal pollution. We establish a bioassay using the quantitative evaluation of metallothionein (MT) transcript in the fish hepatoma cell line, RTH-149, as a tool for detecting heavy metal pollution in brackish-marine water containing other pollutants in addition to heavy metals. RT-competitive polymerase chain reaction was used for the quantitative evaluation of the transcript in absolute units. Cadmium was used as a model pollutant to optimize two parameters of the assay: exposure periods (24, 96, 144 h) and salinity (0%, 25%, 50%, 75%, 100% sea water). Results revealed that salinity at or below 25% sea water at an exposure period of 144 h are the preferable conditions for detecting MT mRNA levels for in vitro assays employed on water samples from highly polluted brackish habitats.
Kamer
I
I
National Institute of Oceanography, Israel Oceanographic and Limnological Research, Tel-Shikmona, P.O.B 8030, Haifa 31080, Israel.
Douek
J
J
Tom
M
M
Rinkevich
B
B
D000818:Animals; D001681:Biological Assay; D001682:Biological Availability; D002104:Cadmium / Q000009:adverse effects* / Q000493:pharmacokinetics*; D006528:Carcinoma, Hepatocellular / Q000473:pathology; D002460:Cell Line; D004790:Enzyme Induction; D005399:Fishes; D008113:Liver Neoplasms / Q000473:pathology; D008668:Metallothionein / Q000096:biosynthesis*; D019216:Metals, Heavy / Q000009:adverse effects* / Q000493:pharmacokinetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014873:Water Pollutants / Q000009:adverse effects* / Q000493:pharmacokinetics*
D019216:Metals, Heavy; D014873:Water Pollutants; D002104:Cadmium; D008668:Metallothionein