Global climate change imposes physiological constraints on marine ecosystems that can alter the distribution of intertidal organisms. In one such instance, the native cold-adapted mussel Mytilus trossulus is being replaced along its southern range by the invasive warm-adapted Mytilus galloprovincialis. These blue mussels occur throughout rocky intertidal zones where they are subjected to greatly varying environmental conditions known to induce oxidative stress. We hypothesize that while under acute stress, related Mytilus congeners undergo a shift in redox potential from NADH-fueled respiratory pathways to pathways producing NADPH as a way to decrease the production of reactive oxygen species (ROS) and provide reducing equivalents to detoxify ROS. Additionally, we hypothesize that sirtuins (SIRT; a family of NAD-dependent deacetylases) might be involved in the regulation of this metabolic transition. To test the latter, a discovery approach will be used to analyze the proteomic response of M. galloprovincialis and M. trossulus to the pro-oxidant menadione, and sirtuin-inhibitors nicotinamide and suramin. Menadione can induce oxidative stress by increasing endogenous peroxide and superoxide radicals, while suramin and nicotinamde both inhibit sirtuin activity. Organisms were exposed to these compounds in filtered seawater for 8 h, followed by a 24.5 h recovery period under constant aeration. A multivariate analysis utilizing 2D-gel electrophoresis and protein identification via mass spectrometry showed that 18% and 17% of all identified protein spots detected demonstrated changes in abundance in M. galloprovincialis and M. trossulus, respectively. Using matrix-assisted laser desorption ionization (MALDI) tandem time-of-light mass spectrometry, we were able to identify 32-41% of proteins, depending on the species.
The two Mytilus congeners showed the greatest differences in changes of protein abundance for oxidative stress proteins (including NADP-dependent isocitrate dehydrogenase). Both congeners showed similar effects in response to simultaneous sirtuin inhibition and MIOS for proteins involved in protein degradation (proteasome), cytoskeletal modifications (actin and tubulin), proteins regulating actin filament growth (F-actin capping protein), amino acid metabolism and stress signaling (G-proteins, small G-proteins and MAPK). Results indicate that protein acetylation plays an important role in the oxidative stress response of M. galloprovincialis. More specifically this suggests that sirtuins play an important role in regulating the general stress response in M. galloprovincialis and thus contribute to the greater stress resistance of this species. Furthermore, changes in the abundance of several molecular chaperones suggest a greater effect of sirtuins in regulating the cellular response to heat stress, which could in part explain why this species is more heat-tolerant than the native M. trossulus.
| Identifer | oai:union.ndltd.org:CALPOLY/oai:digitalcommons.calpoly.edu:theses-2436 |
| Date | 01 June 2014 |
| Creators | Chilton, Hayley C |
| Publisher | DigitalCommons@CalPoly |
| Source Sets | California Polytechnic State University |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Master's Theses |
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