Graduate Thesis Or Dissertation


Exploring the cellular mechanisms of Cnidarian bleaching in the sea anemone Aiptasia pallida Public Deposited

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  • Many members of the Phylum Cnidaria are mutualistic with unicellular dinoflagellates belonging to the genus Symbiodinium. Corals are the most widely recognized example of these associations due to their key ecological importance in coral reef ecosystems where they serve as the structural and trophic foundation of these rich ecosystems. Coral reefs are severely threatened by human activities worldwide and are at great risk from global climate change, in particular the increase in seasurface temperatures. Detailed knowledge of how corals respond to stress is scarce. The most serious and immediate response of corals to environmental stress is a process referred to as coral bleaching (a.k.a. cnidarian bleaching). Nevertheless, the cellular and molecular processes by which elevated temperatures elicit the bleaching response are poorly understood. This dissertation deals with this important question by describing two mediators of cnidarian bleaching in the model symbiotic tropical sea anemone Aiptasia pallida (Verril), namely nitric oxide and cyclophilin. After an introduction to the topic of cnidarian-algal symbioses and cnidarian bleaching (Chapter 1), I present results from a study describing the involvement of nitric oxide (NO) in the anemone A. pallida (Chapter 2). Elevated temperature as well as oxidative stress induces production of NO and exposure of A. pallida to NO induces bleaching at non-stressful temperatures. Co-incubation with an NO scavenger suppresses bleaching. I propose that the host up-regulates NO production in response to elevated oxidative stress and that this situation leads to cytotoxicity and bleaching. Chapter 3 examines the role of cyclophilin from A. pallida in the regulation of the symbiosis. Cyclophilins belong to a highly conserved family peptydyl-prolyl cistrans isomerases (PPIases). Incubation of A. pallida with cyclosporin A (CsA), a potent inhibitor of cyclophilin resulted in bleaching and a decrease in tolerance to elevated temperatures. Protein extracts from A. pallida exhibited CsA-sensitive PPIase activity. Laser scanning confocal microscopy using superoxide and nitric oxide-sensitive fluorescent dyes on live A. pallida revealed that CsA strongly induced the production reactive oxygen species as well as NO. We tested weather the CsAsensitive isomerase activity is important for maintaining the activity of the antioxidant enzyme superoxide dismutase (SOD). SOD activity of protein extracts was not affected by pre-incubation with CsA in vitro. In Chapter 4 I review what is known about the molecular and cellular mechanisms of bleaching and describe a model of bleaching based on the results presented herein as well as studies of non-cnidarian models.
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