Scientif poster I gonna present at the SETAC Europe annual meeting in Seville, Spain. May 2010Weniger lesen
![WAP65 IS REDUCED DURING CADMIUM ACCLIMATION IN LEAST KILLIFISH, HETERANDRIA FORMOSA , AS REVEALED BY SUB-PROTEOMIC ANALYSIS Frédéric SILVESTRE 1 , Joseph ADEYEMI 2 , Marc DIEU 3 , Martine RAES 3 , Patrick KESTEMONT 1 , Paul KLERKS 2 This program is supported by the « Funds for Scientific Research FNRS-FRS » - Belgium [email_address] + 153% + 28% Fig. 1. Median TTD (hours ± 25% quantiles) of H. formosa exposed to 6 mg/L Cd after an acclimation period of 1, 2 or 3 weeks (1w, 2w, 3w) and a decontamination period of 0 or 3 weeks. Cd concentration of 0, 1, 5 or 10 µg/L during the acclimation period. + 16% ,[object Object],[object Object],[object Object],[object Object],Time to Death (TTD) after the acclimation and/or decontamination period, 20 fish were exposed to a lethal Cd level of 6 mg/L median time-to-death (TTD) was calculated with STATISTICA ™using t he logrank test to compare the survival distribution of one exposed group with the control group. DIGE analysis (Differential in Gel Electrophoresis) fifty 1 to 7-day old fish in thirty six 10-gallon glass tanks filled with 30 L of dechlorinated water acclimation period of 1, 2 or 3 weeks with Cd at 0, 1, 5 or 10 µg/L decontamination period of 0, 1 or 3 weeks (with no Cd) Acclimation experiment ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],TTD TTD TTD TTD TTD TTD Acclimation 1 week Acclimation 2 weeks Acclimation 3 weeks Decontamination 0 or 3 weeks Least killifish were sampled in Lake Martin (right picture) near Lafayette, Louisiana, in shallow water swamp. 2 weeks exposure to 1 µg Cd/L vs. control 3 weeks decontamination – Fraction S 2 weeks exposure to 1 µg Cd/L vs. control 3 weeks decontamination – Fraction MLP 18 protein spots (p<0.05) : 9 over-expressed (1.2 – 1.9-fold) 9 under-expressed (1.2 – 1.8-fold) 65 protein spots (p<0.05) : 30 over-expressed (1.1 – 1.6-fold) 35 under-expressed (1.1 – 2.1-fold) Fig. 2. Typical 2D gels of fractions S (left) and MLP (right) proteins from Heterandria formosa . Spots over or under-expressed in fish acclimated to 1 µg/L Cd for 2 weeks followed by 3 weeks decontamination, compared to their respective unexposed controls. P<0.05. N=4. Fraction M r (kDa) p I (pH units) Protein name Species Accession No. No. of identified peptides Sum of Mascot score Best ion score Expected value Identified peptide sequence Fold change p- value S 49.8 6.4 Warm temperature-acclimated- related 65kDa protein (Wap65) Oryzias latipes Q8JIP9 2 185 87 0.0000021 VSLELFGCDH -1.78 0.018 VHLDAITSDDAGNIYAFR S 49.8 6.4 Warm temperature-acclimated- related 65kDa protein (Wap65) Oryzias latipes Q8JIP9 3 265 87 0.0000021 VSLELFGCDH -1.57 0.033 VHLDAITSDDAGNIYAFR SFAELDDHHHLGHVDAA FR MLP 39.7 7.5 Arrestin Tetraodon nigroviridis Q4SVF7 3 326 123 0.0000000069 YGSEDLDVMGLCFR 1.15 0.023 LKDEDTNLASTTMLR KIQYAPSQVGAGPK MLP 54.5 5.4 Protein disulfide isomerase family Tetraodon nigroviridis Q4SE84 3 260 97 0.0000025 VVVAENFDSIVNDDSK 1.13 0.041 YGVSGYPTLK LAPEYEAAATR Table 1. Identification of 4 protein spots by MS/MS. Accession No. are from Uniprot. Conclusions Introduction The least killifish Heterandria formosa is a small ovoviviparous fish living in freshwater along the coast from Louisiana to North Carolina. It can adapt as quickly as the second generation following selection by lethal Cd exposure. Usually, organisms chronically exposed to a pollutant can develop an increased resistance to this chemical during an acclimation process, which is due to some phenotypic plasticity. However, mechanisms underlying this phenomena are largely unknown and could provide new insight into acclimation and detoxification process. Objectives The main objectives were 1) to determine the acclimation window, i.e. the chronic exposure conditions (time and concentration) to Cd that increase the resistance to a subsequent lethal exposure ; 2) to determine molecular mechanisms underlying acclimation, using a sub-proteomic approach. 1. Unité de Recherche en Biologie des Organismes, the University of Namur FUNDP, 61 rue de Bruxelles, 5000 Namur, Belgium. 2. Department of Biology, the University of Louisiana at Lafayette, Lafayette, LA, USA. 3. Unité de Recherche en Biologie Cellulaire, the University of Namur FUNDP, 61 rue de Bruxelles, 5000 Namur, Belgium. Results ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]](https://image.slidesharecdn.com/postersville2010fs2-12743845560514-phpapp01/85/Poster-SETAC-Seville-2010-1-320.jpg)
