oyster hemocytes - nsa 2009
DESCRIPTION
TRANSCRIPT
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Analysis of Genes Isolated from Plated Hemocytes
of the Pacific Oyster, Crassostrea gigas
Steven RobertsSchool of Aquatic and Fishery Sciences
University of Washington
Rick Goetz Great Lakes WATER Institute
University of Wisconsin
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Goal
Better understand stress response
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Laura Canesi, Gabriella Gallo, Miriam Gavioli, and Carla PruzzoBacteria–hemocyte Interactions and Phagocytosis in MarineBivalves. 2002. Microscopy Research And Technique 57:469–476
Immune System
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Specific Objectives
• Identify genes expressed in oyster hemocytes
• Characterize response of select molecules following Vibrio challenge
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Outline
•cDNA Library Description
•Select Functional Categories• Proteases and Protease Inhibitors
• Receptor and Associated Proteins
• Transcription/Translation/Cell Cycle
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Pacific OysterC. gigas
Incubate 24 hours Wash in sterile seawaterExtract with Tri-Reagent
+ sterilesaltwater
Plated hemocyte cDNA library
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Plated hemocyte cDNA library
Prior to washing After washing
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Gene - DNA
Transcript - mRNA
Extract from tissue
Make cDNA
Clone into a “Vector”
Randomly sequence clones = “Expressed Sequence Tags”
(ESTs)
ESTs
(Complementary DNA = Copy)
= cDNA Library
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C. gigas plated hemocytes
Roberts et al 2009
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C. gigas plated hemocytes
modified from Roberts et al 2009
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Markers &more
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Markers &more
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C. gigas Vibrio challenge
Roberts et al 2009
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Proteases and AntiproteasesGene Function
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Proteases and AntiproteasesGene Function
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Proteases and AntiproteasesGene Function
Inhibitors
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Proteases and AntiproteasesGene Function
Cathepsins
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Proteases and AntiproteasesGene Function
Cathepsins
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Proteases and AntiproteasesGene Function
C. virginica Homologs
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Proteases and AntiproteasesGene
C. virginica Homologs
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Receptors and other proteinsGene Function
Prostaglandin E2EP4 subtype
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Prostaglandin
15-hydroxyprostaglandin dehydrogenase
PGE2 breakdown
lipoxygenases production of leukotrienes,
lipoxins,hydroperoxy fatty acids
other ESTs present
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Prostaglandin
Mackenzie Gavery - Poster
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Receptors and other proteinsGene Function
Ectodysplasin A2Receptor
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Transcription / Translation / Cell CycleGene Function
Erythroid differentiation-related factor
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Transcription / Translation / Cell CycleGene Function High-mobility
group proteins
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Transcription / Translation / Cell CycleHigh-mobilitygroup proteins
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Transcription / Translation / Cell CycleHigh-mobilitygroup proteins
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Conclusions
• Several novel genes were identified from C. gigas hemocytes
• Hemocyte signaling plays important role in immune response
• Stress response is diverse and complex
•
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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