Aquaculture Research

Steven RobertsAssistant Professor

Genomic approaches in

Friday, May 15, 2009

Overview

Big Fish

Better Shellfish

Bad Bugs

Myostatin Regulation

Disease Resistance

Pathogen Virulence

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Myostatin

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Does myostatin function in a similar manner in fish as it does in mammals?

=

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Approach

LAP

Identify molecules that will decrease biological activity

MSTN (mature peptide)

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Identification of Myostatin Interacting Proteins

Approach: Yeast two hybrid system

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Yeast-two-Hybrid

1.

2.

Telethonin

LAP

Results

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Future Work

• Co-IP targets identified with Y2H

• Sequence proteins from muscle extract captured via co-IP

• Primary muscle culture experiments

• Large scale recombinant protein expression and expose juvenile fish

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Acknowledgements

FUNDING

This project is supported by National Research

Initiative Competitive Grant no. 2005-35206-15261

from the USDA Cooperative State Research,

Education, and Extension Service.

Technical AssistanceChristina Holmes

Sam White

Friday, May 15, 2009

Overview

Big Fish

Better Shellfish

Bad Bugs

Myostatin Regulation

Disease Resistance

Pathogen Virulence

OystersAbalone

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MVineyardflickr | cpurrin1

Disease Resistance ?

Roxannna SmolowitzRick Karney

Crassostrea virginica

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•Mortality

•Disease

•Immune Response

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C. virginica

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Abalone

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Glenn VanBlaricom, Carolyn Friedman

Disease Pressure

Naive

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Carolyn Friedman

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H. cracherodii

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H. cracherodii

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Summary

• Just as with oysters, processes of natural selection have resulted in populations with increased tolerance.

• Important mechanisms involved in the immune response to WS are associated with initial pathogen recognition.

Friday, May 15, 2009

Overview

Big Fish

Better Shellfish

Bad Bugs

Myostatin Regulation

Disease Resistance

Pathogen Virulence

Vibrio tubiashii

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How do changes in the environment influence Vibrio tubiashii physiology?

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Approach - Protein Expression

Protein Extraction

2D gels

ID differences

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Differential Expression

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biosynthesis

response to endogenous stimulus

transport

ion transport

signal transduction

cell communication

transcription

protein metabolism

cell cycle

carbohydrate metabolism

protein biosynthesis

cell organization and biogenesis

response to external stimulus

amino acid and derivative metabolism

morphogenesis

protein modification

generation of precursor metabolites and energy

death

response to abiotic stimulus

cell death

cell differentiation

cell homeostasis

nucleic acid metabolism

secondary metabolism

catabolism

lipid metabolism

DNA metabolism

response to stress 0 0.025 0.050 0.075 0.100

Chart 6

Low Oxygen

Control

GO Annotations

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V. t

ubia

shii

V. t

ubia

shii

+

live

oyst

ers

V. t

ubia

shii

+

ster

ile o

yste

rs

Oys

ters

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pI MW3.7 29k4.5 49k4.8 26k M

W

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Vibrio tubiashii

226 million bp

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Future Direction

• Comparative microbial analysis to determine species specific responses to environmental change

• Coordinate with experimental and environmental sampling - relevance

• Characterize the host response in relation to pathogen physiology

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Thank you

for more information

fish.washington.edu/genefishsr320@washington.edu

Friday, May 15, 2009