Oyster Hemocytes - NSA 2009

Analysis of Genes Isolated from Plated Hemocytes of the Paciﬁc Oyster, Crassostrea gigas Steven Roberts School of Aquatic and Fishery Sciences University of Washington Rick Goetz Great Lakes WATER Institute University of Wisconsin

Goal Better understand stress response

Immune System Laura Canesi, Gabriella Gallo, Miriam Gavioli, and Carla Pruzzo Bacteria–hemocyte Interactions and Phagocytosis in Marine Bivalves. 2002. Microscopy Research And Technique 57:469–476

Speciﬁc Objectives • Identify genes expressed in oyster hemocytes • Characterize response of select molecules following Vibrio challenge

Outline • cDNA Library Description • Select Functional Categories • Proteases and Protease Inhibitors • Receptor and Associated Proteins • Transcription/Translation/Cell Cycle

Plated hemocyte cDNA library Incubate 24 hours Pacific Oyster Wash in sterile seawater C. gigas Extract with Tri-Reagent + sterile saltwater

Plated hemocyte cDNA library Prior to washing After washing

Extract from tissue ESTs Make cDNA Gene - DNA (Complementary DNA = Copy) Clone into a “Vector” Transcript - mRNA = cDNA Library Randomly sequence clones = “Expressed Sequence Tags” (ESTs)

C. gigas plated hemocytes Roberts et al 2009

C. gigas plated hemocytes modiﬁed from Roberts et al 2009

Markers & more

C. gigas Vibrio challenge Roberts et al 2009

Proteases and Antiproteases Gene Function

Proteases and Antiproteases Gene Function Inhibitors

Proteases and Antiproteases Gene Function Cathepsins

Proteases and Antiproteases Gene Function C. virginica Homologs

Proteases and Antiproteases Gene C. virginica Homologs

Receptors and other proteins Gene Function Prostaglandin E2 EP4 subtype

Prostaglandin other ESTs present lipoxygenases production of leukotrienes, lipoxins, hydroperoxy fatty acids 15-hydroxyprostaglandin dehydrogenase PGE2 breakdown

Prostaglandin Mackenzie Gavery - Poster

Receptors and other proteins Gene Function Ectodysplasin A2 Receptor

Transcription / Translation / Cell Cycle Gene Function Erythroid differentiation- related factor

Transcription / Translation / Cell Cycle Gene Function High-mobility group proteins

Transcription / Translation / Cell Cycle High-mobility group proteins

Conclusions • Several novel genes were identiﬁed from C. gigas hemocytes • Hemocyte signaling plays important role in immune response • Stress response is diverse and complex •

Conclusions • Several novel genes were identiﬁed from C. gigas hemocytes • Hemocyte signaling plays important role in immune response • Stress response is diverse and complex • h/t

Oyster Hemocytes - NSA 2009

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