Clone-specific differences and dynamics of abiotic stress effects on the Daphnia pulex proteome

Daphnia pulex unterliegt in ihrem Habitat starken Futter- und Temperatur-Schwankungen. Veränderungen der Protein-Expression innerhalb des Proteoms während akutem Hunger-, Hitze- und kombiniertem Hunger- und Hitze-Stress wurden in dieser Arbeit in zwei unterschiedlichen D. pulex-Klonen durch 2D-Gelel...

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Weiterer Titel:Clone specific differences and dynamics of abiotic stress effects on the Daphnia pulex proteome
Verfasser: Reydelet, Yann
Weitere Beteiligte: Paul, Rüdiger J. (Gutachter)
Dokumenttypen:Dissertation/Habilitation
Medientypen:Text
Erscheinungsdatum:2015
Publikation in MIAMI:11.01.2016
Datum der letzten Änderung:07.04.2022
Angaben zur Ausgabe:[Electronic ed.]
Schlagwörter:Daphnia pulex; Proteom; 2D-Gelelektrophorese; Massenspektrometrie; Hitzestress; zelluläre Stressantwort; Überleben; Proteinexpression; Gesamtproteingehalt; Hungerstress Daphnia pulex; 2D gel electrophoresis; starvation; heat-stress; mass spectrometry; survival; protein expression; total protein; cellular stress response; proteom
Fachgebiet (DDC):570: Biowissenschaften; Biologie
Lizenz:InC 1.0
Sprache:English
Format:PDF-Dokument
URN:urn:nbn:de:hbz:6-17229678635
Permalink:https://nbn-resolving.de/urn:nbn:de:hbz:6-17229678635
Onlinezugriff:diss_reydelet.pdf
Inhaltsverzeichnis:
  • Content I
  • List of Figures III
  • List of Tables V
  • List of Abbreviations VI
  • Abstract 1
  • 1 Background 5
  • 1.1 Stress – Response and Adaptation 5
  • 1.2 Daphnia: a model organism 11
  • 1.3 Clonal variation and local adaptation 13
  • 1.4 Effects of changes in temperature on organisms and their coping mechanisms 17
  • 1.5 Adaptation to the change of food availability 19
  • 1.6 Goals of the thesis 22
  • 2 Methods 23
  • 2.1 Animals 23
  • 2.2 Clone identification by allozyme characterization 23
  • 2.3 Motility and survival assay 23
  • 2.4 Long term swimming assay 24
  • 2.5 Stress exposure and protein extraction 24
  • 2.6 Evaluation of total protein amount 25
  • 2.7 Two-dimensional gel electrophoresis 25
  • 2.8 Analysis of differential protein expression 26
  • 2.9 LC-MS/MS, identification and characterization of proteins 27
  • 2.10 HSP60 quantification with Western Blots 28
  • 3 Results 31
  • 3.1 Clonal characterization by allozyme analysis 31
  • 3.2 Temporal changes in motility and long-term survival under control and stress
  • conditions 32
  • 3.3 Protein expression at different environmental conditions 35
  • 3.3.1 Total protein quantities at different environmental conditions 35
  • 3.3.2 Protein expression at control conditions (20 °C, ad libitum food supply) 36
  • 3.3.3 Protein expression at heat stress (30 °C, ad libitum food supply) 38
  • 3.3.4 Protein expression in two D. pulex clones at starvation stress 51
  • 3.3.5 Protein expression at heat-and-starvation stress (30 °C, no food) 61
  • 3.3.6 Summary of differential protein expressions 71
  • 3.4 Validation of the protein expression changes: HSP60 74
  • 4 Discussion 79
  • 4.1 Major differences in stress-induced proteomic responses between the D. pulex clones G and M 81
  • 4.2 Differences in heat-induced proteomic responses between the fed and non-fed D. pulex clones G and M 84
  • 4.3 Differences in starvation-induced proteomic responses between the D.pulex clones G and M at 20°C and 30°C 90
  • 4.4 Stress responses in the D.pulex clones G and M: conclusions and short outlook 95
  • 4.5 Validation of the protein pattern by Western Blot analysis 96
  • 4.6 Comparison of transcriptome and proteome 98
  • 5 Supplement 99
  • 5.1 Validation of the protein expression changes: HSP60 99
  • 5.2 Validation of the protein pattern by Western Blot analysis 106
  • 5.3 Regulation of gene expression 109
  • 5.4 Regulation of gene expression via core promoters 110
  • 5.5 Bioinformatical analysis for specific promoters and highly conserved protein
  • domains 115
  • 5.6 Bioinformatic analysis of promotor regions and enhancers 118
  • 5.7 Bioinformatic analysis of promotor and enhancer regions 129
  • References 135
  • .

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