Using NMR metabolomics to identify responses of an environmental estrogen in blood plasma of fish.

Nuclear magnetic resonance (NMR) based metabolomics in combination with multivariate data analysis may become valuable tools to study environmental effects of pharmaceuticals and other chemicals in aquatic organisms. To explore the usefulness of this approach in fish, we have used 1H NMR metabolomics to compare blood plasma and plasma lipid extracts from rainbow trout exposed to the synthetic contraceptive estrogen ethinylestradiol (EE2) with plasma from control fish. The plasma metabolite profile was affected in fish exposed to 10 ng/L but not 0.87 ng/L of EE2, which was in agreement with an induced vitellogenin synthesis in the high dose group only, as measured by ELISA. The main affected metabolites were vitellogenin, alanine, phospholipids and cholesterol. The responses identified by this discovery-driven method could be put in context with previous knowledge of the effects of estrogens on fish. This adds confidence to the approach of using NMR metabolomics to identify environmental effects of pharmaceuticals and other contaminants.

[1]  Thomas H Hutchinson,et al.  Ecotoxicogenomics: the challenge of integrating genomics into aquatic and terrestrial ecotoxicology. , 2004, Aquatic toxicology.

[2]  D. Tocher,et al.  Influence of plasma lipid changes in response to 17β‐oestradiol stimulation on plasma growth hormone, somatostatin, and thyroid hormone levels in immature rainbow trout , 2001 .

[3]  K. Oakes,et al.  Short‐term lab exposures of immature rainbow trout (Oncorhynchus mykiss) to sulfite and kraft pulp‐mill effluents: Effects on oxidative stress and circulating sex steroids , 2005, Environmental toxicology and chemistry.

[4]  Sunia A Trauger,et al.  Metabonomic assessment of toxicity of 4‐fluoroaniline, 3,5‐difluoroaniline and 2‐fluoro‐4‐methylaniline to the earthworm Eisenia veneta (rosa): Identification of new endogenous biomarkers , 2002, Environmental toxicology and chemistry.

[5]  A. Cantafora,et al.  The influence of estrogen on hepatic cholesterol metabolism and biliary lipid secretion in rats fed fish oil. , 1999, Biochimica et biophysica acta.

[6]  R. Gonzalez,et al.  Quantitative neuropathology by high resolution magic angle spinning proton magnetic resonance spectroscopy. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[7]  M. Casu,et al.  NMR lipid profiles of cells, tissues and body fluids. I— 1D and 2D Proton NMR of lipids from rat liver , 1991 .

[8]  Bruce R. Kowalski,et al.  Chemometrics, mathematics and statistics in chemistry , 1984 .

[9]  D. P. Lomax,et al.  Purification and partial characterization of English sole (Pleuronectes vetulus) vitellogenin. , 1997, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[10]  J. Sumpter,et al.  Estrogenic Effects of Effluents from Sewage Treatment Works , 1994 .

[11]  N. Woo,et al.  Influence of oral administration of estradiol-17β and testosterone on growth, digestion, food conversion and metabolism in the underyearling red sea bream, Chrysophrys major , 1993, Fish Physiology and Biochemistry.

[12]  Ronald S. Tjeerdema,et al.  NMR-derived developmental metabolic trajectories: an approach for visualizing the toxic actions of trichloroethylene during embryogenesis , 2005, Metabolomics.

[13]  D. Larsson,et al.  Seasonal variations of vitelline envelope proteins, vitellogenin, and sex steroids in male and female eelpout (Zoarces viviparus). , 2002, General and comparative endocrinology.

[14]  D. Leibfritz,et al.  Assignment of mono‐ and polyunsaturated fatty acids in lipids of tissues and body fluids , 1998 .

[15]  H. Cooper,et al.  Liver tumors in wild flatfish: a histopathological, proteomic, and metabolomic study. , 2005, Omics : a journal of integrative biology.

[16]  J. Griffin,et al.  High-resolution magic angle spinning 1H-NMR spectroscopy studies on the renal biochemistry in the bank vole (Clethrionomys glareolus) and the effects of arsenic (As3+) toxicity , 2001, Xenobiotica; the fate of foreign compounds in biological systems.

[17]  S. Meiboom,et al.  Modified Spin‐Echo Method for Measuring Nuclear Relaxation Times , 1958 .

[18]  S. Jørgensen,et al.  Occurrence, fate and effects of pharmaceutical substances in the environment--a review. , 1998, Chemosphere.

[19]  Marilyn J. Aardema,et al.  Toxicology and genetic toxicology in the new era of "toxicogenomics": impact of "-omics" technologies. , 2002 .

[20]  C. Lazier,et al.  Estrogen responsiveness of vitellogenin gene expression in rainbow trout (Oncorhynchus mykiss) kept at different temperatures. , 1993, General and comparative endocrinology.

[21]  P. J. Babin,et al.  Effects of 17β-estradiol and starvation on trout plasma lipoproteins , 1992, Lipids.

[22]  E. Thurman,et al.  Pharmaceuticals, hormones, and other organic wastewater contaminants in U.S. streams, 1999-2000: a national reconnaissance. , 2002 .

[23]  J N Lester,et al.  The Effects of Natural and Synthetic Steroid Estrogens in Relation to their Environmental Occurrence , 2002, Critical reviews in toxicology.

[24]  C. Haux,et al.  Isolation, immunochemical detection, and observations of the instability of vitellogenin from four teleosts , 1993 .

[25]  D. Cowburn,et al.  Correlation of 13C and 1H chemical shifts in bovine high-density lipoprotein from two-dimensional NMR , 1984 .

[26]  Ceri A. Morris,et al.  Dynamics of estrogen biomarker responses in rainbow trout exposed to 17β‐estradiol and 17α‐ethinylestradiol , 2003 .

[27]  J. Lindon,et al.  Environmental Metabonomics: Applying Combination Biomarker Analysis in Earthworms at a Metal Contaminated Site , 2004, Ecotoxicology.

[28]  John C. Lindon,et al.  Metabonomics: metabolic processes studied by NMR spectroscopy of biofluids , 2000 .

[29]  John C Lindon,et al.  An integrated metabonomic investigation of acetaminophen toxicity in the mouse using NMR spectroscopy. , 2003, Chemical research in toxicology.

[30]  Robert S Plumb,et al.  Metabonomics: the use of electrospray mass spectrometry coupled to reversed-phase liquid chromatography shows potential for the screening of rat urine in drug development. , 2002, Rapid communications in mass spectrometry : RCM.

[31]  M. Viant,et al.  Characterizing the metabolic actions of natural stresses in the California red abalone, Haliotis rufescens using 1H NMR metabolomics , 2005, Metabolomics.

[32]  T. Heberer Occurrence, fate, and removal of pharmaceutical residues in the aquatic environment: a review of recent research data. , 2002, Toxicology letters.

[33]  Ian D. Wilson,et al.  HIGH RESOLUTION PROTON MAGNETIC RESONANCE SPECTROSCOPY OF BIOLOGICAL FLUIDS , 1989 .

[34]  Mark R Viant,et al.  NMR-based metabolomics: a powerful approach for characterizing the effects of environmental stressors on organism health. , 2003, Environmental science & technology.

[35]  Kurt Wüthrich,et al.  Experimental techniques of two-dimensional correlated spectroscopy , 1980 .

[36]  Mika Ala-Korpela,et al.  1H NMR spectroscopy of human blood plasma , 1995 .

[37]  S. Wold,et al.  Partial least squares analysis with cross‐validation for the two‐class problem: A Monte Carlo study , 1987 .

[38]  M. Pettersson,et al.  ETHINYLOESTRADIOL - AN UNDESIRED FISH CONTRACEPTIVE , 1999 .

[39]  J. Rotchell,et al.  Molecular Markers of Endocrine Disruption in Aquatic Organisms , 2003, Journal of toxicology and environmental health. Part B, Critical reviews.

[40]  C. Haux,et al.  Fatty acid composition of vitellogenin from four teleost species , 1995, Journal of Comparative Physiology B.

[41]  M. Pettersson,et al.  Contraceptive pill residues in sewage effluent are estrogenic to fish , 2000 .

[42]  G. Ankley,et al.  Ecotoxicogenomics: linkages between exposure and effects in assessing risks of aquatic contaminants to fish. , 2005, Reproductive toxicology.

[43]  M. Spraul,et al.  750 MHz 1H and 1H-13C NMR spectroscopy of human blood plasma. , 1995, Analytical chemistry.

[44]  M. Scholze,et al.  Relative potencies and combination effects of steroidal estrogens in fish. , 2003, Environmental science & technology.

[45]  S. Wold,et al.  Multivariate Data Analysis in Chemistry , 1984 .