SAFS Aquaculture

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


Myostatin


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

=


Approach

LAP

Identify molecules that will decrease biological activity

MSTN (mature peptide)


Identification of Myostatin Interacting Proteins

Approach: Yeast two hybrid system


Yeast-two-Hybrid

1.

2.

Telethonin

LAP

Results


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


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


Overview

Big Fish

Better Shellfish

Bad Bugs


Disease Resistance

Pathogen Virulence

OystersAbalone


MVineyardflickr | cpurrin1

Disease Resistance ?

Roxannna SmolowitzRick Karney

Crassostrea virginica


•Mortality

•Disease

•Immune Response


C. virginica


Abalone


Glenn VanBlaricom, Carolyn Friedman

Disease Pressure

Naive


Carolyn Friedman


H. cracherodii


H. cracherodii


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.


Overview

Big Fish

Better Shellfish

Bad Bugs


Disease Resistance

Pathogen Virulence

Vibrio tubiashii


How do changes in the environment influence Vibrio tubiashii physiology?


Approach - Protein Expression

Protein Extraction

2D gels

ID differences


Differential Expression


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


V. t

ubia

shii

V. t

ubia

shii

+

live

oyst

ers

V. t

ubia

shii

+

ster

ile o

yste

rs

Oys

ters


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

W


Vibrio tubiashii

226 million bp


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


Thank you

for more information

fish.washington.edu/[email protected]


Business

SAFS Aquaculture