Upload
mgavery
View
282
Download
1
Embed Size (px)
DESCRIPTION
School of Aquatic & Fishery Sciences Graduate Student Symposium
Citation preview
Characterization of prostaglandins in the Pacific oyster Crassostrea gigas: evidence for a role in immune response
Mackenzie Gavery, M.S. StudentRoberts Lab
Molecules with Diverse Functions•Vertebrates
• Mediate swelling and inflammation
• (COX inhibitors)
• Inducing fever
• Maintaining pregnancy/reproductive timing
• Regulating ion transport
Molecules with Diverse Functions
• Invertebrates
• Ion transport
• Reproduction
• Immune Response
•recently discovered a gene encoding prostaglandin E2 receptor in Pacific oysters
• significantly upregulated after exposure to Vibrio tubiashii
Roberts et al
What are prostaglandins?•Eicosanoids: a family of biologically active
metabolites of C20 polyunsaturated fatty acids.
•Prostaglandins biosynthesis begins with the from arachidonic acid, an omega-6 fatty acid.
prostaglandin E2:
Arachidonic Acid Cascade
PGE receptor
Outline•Characterize molecules in prostaglandin
pathway
▫ID gene fragments using ESTs
▫Tissue distribution of oyster
▫Isolate complete ORF of oyster COX gene
•Challenge Experiments
•Applied research
Arachidonic Acid Cascade
PGE receptor
Sequences
PGE receptor
ESTs: snail, mussel
ESTs: oyster!,mussel•AM856036
ESTs: ongoing
ESTs: •EW777722
Data
PGE receptor
Full-length Sequence
PGE receptor
STATS:
•Size: 1700 bp
•pred. # introns: 10
•Amino acids: 587
COX Gene Similarities
Outline•Characterize molecules in prostaglandin
pathway
▫ID gene fragments from genomics database
▫Tissue distribution of oyster
▫Isolate complete ORF or oyster COX gene
•Challenge Experiments
•Applied research
ExperimentsVibrio tubiashii exposures
whole organism plated
hemocytes
Analytical Results
PGE receptor
• PGE (ELISA)
• Receptor (PCR)
Vibrio exposure-ELISA results
PGE conjugate
PGE sample
100% 0%
Gill Tissue/Vibrio– qPCR PGE receptor
Plated hemocytes/Vibrio - qPCR
Outline
•What are prostaglandins?
•Characterization of genes involved in biosynthesis of prostaglandins
•Experiments and Results
•Applied research
Applied Research
Physiological Processes:•Immune Response•Ion transport•Reproduction•Homeostasis
temperature
pathogen exposureacidification
pesticides
Applied Research
•Prostaglandins appear to play a role in the immune response of oysters.
•In vitro hemocyte cultures may be a good experimental model to study physiological responses to disruptions of this pathway
▫how would changes in environmental conditions (pollutants, temperature, pH) affect the regulation of arachidonic acid metabolism?
▫What physiological effects might this have?
Acknowledgements
•Advisor, Steven Roberts•Research scientist, Sam
White•Friedman and Seeb Lab