TY - JOUR
T1 - Quantitative PCR analysis of CYP1A induction in Atlantic salmon (Salmo salar)
AU - Rees, Christopher B.
AU - McCormick, Stephen D.
AU - Vanden Heuvel, John P.
AU - Li, Weiming
N1 - Funding Information:
Gratitude is extended to Phil Hulbert and the staff at Adirondack Fish Hatchery for donation of hatchery salmon for this project. Dr Kim Scribner generously offered lab space and densitometric equipment. Processing of RNA samples during quantitative PCR analysis would not have been possible without the diligent work of Linda Ferkey. A great thanks goes to Bradley Young for assistance in statistical analysis. Technical suggestions and discussions were given by Scot Libants. Amy Moeckel, Mike O'Dea, Darren Lerner and Junya Hiroi helped in capturing and processing Atlantic salmon captured in the wild. Funding for this project was provided by the Jaqua Foundation and Michigan State University.
PY - 2003/1/10
Y1 - 2003/1/10
N2 - Environmental pollutants are hypothesized to be one of the causes of recent declines in wild populations of Atlantic salmon (Salmo salar) across Eastern Canada and the United States. Some of these pollutants, such as polychlorinated biphenyls and dioxins, are known to induce expression of the CYP1A subfamily of genes. We applied a highly sensitive technique, quantitative reverse transcription-polymerase chain reaction (RT-PCR), for measuring the levels of CYP1A induction in Atlantic salmon. This assay was used to detect patterns of CYP1A mRNA levels, a direct measure of CYP1A expression, in Atlantic salmon exposed to pollutants under both laboratory and field conditions. Two groups of salmon were acclimated to 11 and 17°C, respectively. Each subject then received an intraperitoneal injection (50 mg kg-1) of either β-naphthoflavone (BNF) in corn oil (10 mg BNF ml-1 corn oil) or corn oil alone. After 48 h, salmon gill, kidney, liver, and brain were collected for RNA isolation and analysis. All tissues showed induction of CYP1A by BNF. The highest base level of CYP1A expression (2.56×1010 molecules/μg RNA) was found in gill tissue. Kidney had the highest mean induction at five orders of magnitude while gill tissue showed the lowest mean induction at two orders of magnitude. The quantitative RT-PCR was also applied to salmon sampled from two streams in Massachusetts, USA. Salmon liver and gill tissue sampled from Millers River (South Royalston, Worcester County), known to contain polychlorinated biphenyls (PCBs), showed on average a two orders of magnitude induction over those collected from a stream with no known contamination (Fourmile Brook, Northfield, Franklin County). Overall, the data show CYP1A exists and is inducible in Atlantic salmon gill, brain, kidney, and liver tissue. In addition, the results obtained demonstrate that quantitative PCR analysis of CYP1A expression is useful in studying ecotoxicity in populations of Atlantic salmon in the wild.
AB - Environmental pollutants are hypothesized to be one of the causes of recent declines in wild populations of Atlantic salmon (Salmo salar) across Eastern Canada and the United States. Some of these pollutants, such as polychlorinated biphenyls and dioxins, are known to induce expression of the CYP1A subfamily of genes. We applied a highly sensitive technique, quantitative reverse transcription-polymerase chain reaction (RT-PCR), for measuring the levels of CYP1A induction in Atlantic salmon. This assay was used to detect patterns of CYP1A mRNA levels, a direct measure of CYP1A expression, in Atlantic salmon exposed to pollutants under both laboratory and field conditions. Two groups of salmon were acclimated to 11 and 17°C, respectively. Each subject then received an intraperitoneal injection (50 mg kg-1) of either β-naphthoflavone (BNF) in corn oil (10 mg BNF ml-1 corn oil) or corn oil alone. After 48 h, salmon gill, kidney, liver, and brain were collected for RNA isolation and analysis. All tissues showed induction of CYP1A by BNF. The highest base level of CYP1A expression (2.56×1010 molecules/μg RNA) was found in gill tissue. Kidney had the highest mean induction at five orders of magnitude while gill tissue showed the lowest mean induction at two orders of magnitude. The quantitative RT-PCR was also applied to salmon sampled from two streams in Massachusetts, USA. Salmon liver and gill tissue sampled from Millers River (South Royalston, Worcester County), known to contain polychlorinated biphenyls (PCBs), showed on average a two orders of magnitude induction over those collected from a stream with no known contamination (Fourmile Brook, Northfield, Franklin County). Overall, the data show CYP1A exists and is inducible in Atlantic salmon gill, brain, kidney, and liver tissue. In addition, the results obtained demonstrate that quantitative PCR analysis of CYP1A expression is useful in studying ecotoxicity in populations of Atlantic salmon in the wild.
UR - http://www.scopus.com/inward/record.url?scp=12244312781&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=12244312781&partnerID=8YFLogxK
U2 - 10.1016/S0166-445X(02)00062-0
DO - 10.1016/S0166-445X(02)00062-0
M3 - Article
C2 - 12413795
AN - SCOPUS:12244312781
SN - 0166-445X
VL - 62
SP - 67
EP - 78
JO - Aquatic Toxicology
JF - Aquatic Toxicology
IS - 1
ER -