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-Gelelektrophorese und Massenspektrometrie untersucht. Viele der bestimmten Protein-Isoformen waren nach Hunger- und/oder Hitze-Stress in beiden Klonen herauf- oder herunterregeguliert, wobei die Stärke der Expression und die regulatorischen Muster variierten. Glutathion-Transferasen, V-ATPase-Untereinheiten, Zytoskelett-Proteine und Chaperone waren signifikant heraufreguliert, Proteine für Proteolyse, Kohlenhydrat-Bindung oder Metabolismus waren herunterreguliert. Die Expression aller Proteine folgte spezifischen zeitlichen Mustern. Die Verfügbarkeit von Futter zeigten Klon-spezifische Unterschiede im Überleben, dem Gesamtproteingehalt und der Protein-Expression während akutem Hitze-Stress.Daphnia pulex is challenged by severe food and temperature changes in its habitat. To identify cellular adjustments in protein expression during acute starvation, heat-stress and the combination of both stress types; the proteomes of two D. pulex clones were studied by 2D gel electrophoresis and mass spectrometry. Several protein isoforms were up- or down-regulated in both clones during acute heat-stress and/or starvation. The expression varied in intensity and regulatory patterns between both clones. Glutathion transferases, V-ATPase subunits, cytoskeleton-proteins and chaperones were significantly up-regulated, proteins for proteolytic processes and carbohydrate binding or metabolism were down-regulated. The expression of all proteins followed specific temporal patterns. Availability of food resources revealed fundamental clone specific differences in survival, total protein content and protein expression during acute heat-stress

Additional file 8: of The transcriptomic and proteomic responses of Daphnia pulex to changes in temperature and food supply comprise environment-specific and clone-specific elements

Table S2. Labeling design of the microarrays. (PDF 83 kb

Legislative Documents

Also, variously referred to as: Senate bills; Senate documents; Senate legislative documents; legislative documents; and General Court documents

Additional file 7: of The transcriptomic and proteomic responses of Daphnia pulex to changes in temperature and food supply comprise environment-specific and clone-specific elements

Figure S6. Experimental design. (PDF 144 kb

Additional file 3: of The transcriptomic and proteomic responses of Daphnia pulex to changes in temperature and food supply comprise environment-specific and clone-specific elements

Figure S2. Two-dimensional protein gels from the D. pulex clone G under starvation stress. The 2D gels, which are fusion (averaged) images from a varying number (n) of gels (biological replicates, 25-30 animals each), show changes in protein expression in the D. pulex clone G after the acute exposure of control animals (20 °C, ad libitum feeding) (blue spots; n = 5) to (a) 24 h (orange spots; n = 4) or (c) 48 h (orange spots; n = 4) of starvation (T = 20 °C). Red or green spot IDs mark significantly up- or downregulated proteins (t-tests, P < 0.05; see Table 4). The scatter plots show changes in expression level (Vrelative, relative spot volume) between control and starving animals (b, 24 h; d, 48 h) of significantly (large circles and letters) or non-significantly (small circles) up- or down-regulated proteins (data from a or c). Proteins, which were upregulated under starvation stress, are found above the diagonal line. (PDF 126 kb

Additional file 5: of The transcriptomic and proteomic responses of Daphnia pulex to changes in temperature and food supply comprise environment-specific and clone-specific elements

Figure S4. Two-dimensional protein gel from the D. pulex clone G under heat-and-starvation stress. The 2D gel, which is a fusion (averaged) image from a varying number (n) of gels (biological replicates, 25-30 animals each), shows changes in protein expression in the D. pulex clone G after the acute exposure of control animals (20 °C, ad libitum feeding) (blue spots; n = 5) to (a) 24 h (orange spots; n = 5) of heat-and-starvation stress (T = 30 °C, starvation). (Clone G did not survive 48 h of heat-and-starvation stress.) Red or green spot IDs mark significantly up- or downregulated proteins (t-tests, P < 0.05; see Table 5). The scatter plot shows changes in expression level (Vrelative, relative spot volume) between control animals and animals exposed to heat-and-starvation stress (b, 24 h) of significantly (large circles and letters) or non-significantly (small circles) up- or down-regulated proteins (data from a). Proteins, which were upregulated under heat-and-starvation stress, are found above the diagonal line. (PDF 152 kb

Additional file 6: of The transcriptomic and proteomic responses of Daphnia pulex to changes in temperature and food supply comprise environment-specific and clone-specific elements

Figure S5. Two-dimensional protein gels from the D. pulex clone M under heat-and-starvation stress. The 2D gels, which are fusion (averaged) images from a varying number (n) of gels (biological replicates, 25-30 animals each), show changes in protein expression in the D. pulex clone M after the acute exposure of control animals (20 °C, ad libitum feeding) (blue spots; n = 4) to (a) 24 h (orange spots; n = 5) or (c) 48 h (orange spots; n = 7) of heat-and-starvation stress (T = 30 °C, starvation). Red or green spot IDs mark significantly up- or downregulated proteins (t-tests, P < 0.05; see Table 5). The scatter plots show changes in expression level (Vrelative, relative spot volume) between control animals and animals exposed to heat-and-starvation stress (b, 24 h; d, 48 h) of significantly (large circles and letters) or non-significantly (small circles) up- or down-regulated proteins (data from a or c). Proteins, which were upregulated under heat-and-starvation stress, are found above the diagonal line. (PDF 194 kb

Additional file 2: of The transcriptomic and proteomic responses of Daphnia pulex to changes in temperature and food supply comprise environment-specific and clone-specific elements

Figure S1. Two-dimensional protein gels from the 20 °C-acclimated D. pulex clones G and M. The RuBPs-stained 2D gels from control animals (20 °C, ad libitum feeding) of (a) the D. pulex clone G [fusion image from n = 5 gels (biological replicates)] and (b) the D. pulex clone M [fusion image from n = 4 gels (biological replicates, 25-30 animals each)] served as reference for the excision of protein spots for mass spectrometry (pI: 4.5-6, molecular mass: 20-150 or 20-100 kDa). Red characters are spot identifiers (IDs). (PDF 118 kb