Prostaglandins in the Pacific oyster:

Investigations into form and function

Mackenzie Gavery, Sam White, Steven Roberts
University of Washington, School of Aquatic and Fishery Sciences

Project Summary
Plasma Phospholipase A
Prostaglandins are a group of lipid Membrane Cyclooxygenase
compounds that function as powerful COOH
(COX)
Full length sequence
biological signaling molecules. Prostaglandin E2
Prostaglandins are derived from Arachidonic isolated.
Previous research Cyclooxygen
arachidonic acid via a series of enzymatic Acid AA: 588 MW:
has suggested the ase
reactions. Although prostaglandin COOH
67.65 kDa
presence of O
pathways have been well characterized in prostaglandins in O
vertebrates, information on the functional bivalves. ELISA PGH2
role in bivalves is lacking. techniques were O PGE
Synthase PGE2 Receptor
The objectives of this study were to: used to identify COOH
Partial sequence
prostaglandin in obtained for Pacific
1) characterize the molecules involved in
Pacific oyster OH oyster from EST
prostaglandin activity and
hemolymph Prostaglandin E2 (GenBank#:
2) identify functional roles of Prostaglandi
(PGE2) EW777722)
prostaglandins in Pacific oysters. n E2
Receptor

Isolation and Characterization of Molecules

A full length cyclooxygenase sequence was obtained using Rapid RT-PCR analysis indicates expression
Amplification of cDNA Ends based PCR. The deduced amino acid of the PGE2 receptor is highest in gill
sequence is most similar to cyclooxygenase in soft coral. Vertebrate and mantle tissues. The partial
While two forms of cyclooxygenase have been identified COX-1 sequence (EW777722) is most similar
in coral and vertebrates, only one was isolated from the oyster. to the EP4 subtype in Danio rerio
Two partial sequence were identified in (36.2% similarity) and Homo sapiens
databases from other molluscs. RT-PCR (41.6% similarity). The EP4 subtype is
analysis indicates cyclooxygenase involved with
is expressed cAMP signaling.
primarily in gill
and mantle. Vertebrate
COX
COX-2
A

COX B
C. gigas
D. rerio
H. sapiens
C. gigas
COX D. rerio
H. sapiens
C. gigas
D. rerio
H. sapiens
C. gigas
15s- D. rerio
H. sapiens
COX

Immune Function: Vibrio tubiashii Challenges Conclusions

To evaluate the role of prostaglandins in Pacific oyster immune response, Vibrio tubiashii • Isolation of full-length cyclooxygenase
challenges were performed in vivo and in vitro. RT-qPCR was used to evaluate gene sequence in Pacific oyster; first report in
expression of the PGE2 receptor in hemocytes. Circulating PGE2 levels were measured any bivalve species.
via ELISA. PGE2 receptor expression was upregulated after exposure to V. tubiashii
• Partial PGE2 receptor sequence
indicating that prostaglandins are involved in the immune response of Pacific oysters.
identified, regulated upon pathogen
PGE2 Receptor PGE2 in Hemolymph exposure.
• Prostaglandin E2 detected and levels
indicate role in immune system signaling.
• Future work includes developing these
molecules as biomarkers for ecosystem
health.