Vitellogenin of the Chilean flounder Paralichthys adspersus as a biomarker of endocrine disruption along the marine coast of the South Pacific. Part I: induction, purification, and identification.
We sought to provide a useful indicator of the presence of endocrine-disrupting contaminants along the marine coast of the South Pacific using Chilean flounder (Paralichthys adspersus). In light of the lack of information on vitellogenin for this species, we induced, purified, and identified the plasma vitellogenin of Chilean flounder inhabiting the Chilean coast. Vitellogenin (Vg) from Chilean flounder was purified by size exclusion and ion-exchange chromatography using plasma from juvenile males induced by injecting 17beta-estradiol. The Vg was detected by SDS-PAGE and Western blot analyses using an antibody against turbot (Scophthalmus maximus) vitellogenin. These analyses revealed a protein band of 205 kDa and three minor bands of 120, 90, and 68 kDa. These proteins were identified as Vg by means of mass spectrometry (LCQ Duo ESI-IT-MS), matching sequences of tryptic peptides to known sequences for several other fish species. The matches showed the presence of vitellogenin (VgI, VgII, Vg A and Vg B) in Chilean flounder, similar to species such as mummichog (Fundulus heteroclitus), Japanese medaka (Oryzias latipes), and white perch (Morone americana). These results are discussed in terms of identifying Vg in Paralichthys adspersus with the antibody to turbot Vg. Moreover, we compare the molecular size of Vg from Chilean flounder (large) with that of other flatfish species. Finally, we discuss the potential use of this molecule as a biomarker for the presence of xeno-estrogenic compounds along the Chilean coastline.
Leonardi
Maritza
M
PIMEX-ARAUCO Program, Faculty of Natural Sciences and Oceanography, University of Concepción, Concepción, Chile. mleonard@udec.cl.
Vera
Jeanett
J
PIMEX-ARAUCO Program, Faculty of Natural Sciences and Oceanography, University of Concepción, Concepción, Chile.
Tarifeño
Eduardo
E
PIMEX-ARAUCO Program, Faculty of Natural Sciences and Oceanography, University of Concepción, Concepción, Chile.|Department of Zoology, Faculty of Natural Sciences and Oceanography, University of Concepción, Concepción, Chile.
Puchi
Marcia
M
Department of Biochemistry and Molecular Biology, Faculty of Biological Sciences, University of Concepción, Concepción, Chile.
Morín
Violeta
V
Department of Biochemistry and Molecular Biology, Faculty of Biological Sciences, University of Concepción, Concepción, Chile.
D000818:Animals; D015415:Biomarkers / Q000378:metabolism*; D015153:Blotting, Western; D002677:Chile; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D052244:Endocrine Disruptors / Q000032:analysis*; D004784:Environmental Monitoring / Q000379:methods*; D005412:Flatfishes / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry; D010140:Pacific Ocean; D013045:Species Specificity; D014819:Vitellogenins / Q000302:isolation & purification* / Q000378:metabolism*; D014874:Water Pollutants, Chemical / Q000032:analysis*
D015415:Biomarkers; D052244:Endocrine Disruptors; D014819:Vitellogenins; D014874:Water Pollutants, Chemical
J Exp Zool. 1997 Jun 15;278(3):156-66
9181695
J Chromatogr. 1989 Sep 15;478(2):387-97
2600147
Anal Biochem. 1976 May 7;72:248-54
942051
Fish Physiol Biochem. 1995 Feb;14(1):1-13
24197267
Mar Environ Res. 2007 Aug;64(2):128-48
17307251
Gen Comp Endocrinol. 2008 Apr 1;156(2):285-97
18342314
Environ Pollut. 2008 Mar;152(1):138-46
17587471
Arch Environ Contam Toxicol. 2009 Apr;56(3):536-45
18712498
Comp Biochem Physiol A Mol Integr Physiol. 2007 Feb;146(2):200-7
17188011
Bull Environ Contam Toxicol. 2000 Feb;64(2):184-90
10656883
Environ Health Perspect. 1995 Oct;103 Suppl 7:173-8
8593867
Nature. 1970 Aug 15;227(5259):680-5
5432063
Sci Total Environ. 2003 Jun 20;309(1-3):93-103
12798095
Comp Biochem Physiol B Biochem Mol Biol. 2004 Oct;139(2):235-44
15465670
Arch Environ Contam Toxicol. 2009 Apr;56(3):546-56
18769848
Aquat Toxicol. 2002 Oct 2;60(1-2):101-10
12204590
Aquat Toxicol. 2001 Apr;52(2):157-76
11164537
Environ Health Perspect. 1995 Oct;103 Suppl 7:9-15
8593883
Mar Pollut Bull. 2007 Sep;54(9):1315-20
17673263
Mol Reprod Dev. 2004 Sep;69(1):109-16
15278911
Microbiology (Reading). 2000 Oct;146 ( Pt 10):2495-2508
11021925