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Fulkerson Capstone

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  • 1. A Comparison of Growth and Gene Expression in Two Species of Oysters By: Katie Fulkerson Advisor: Steven Roberts By: Katie Fulkerson Advisor: Steven Roberts
  • 2. Oyster History
    • Oyster trade began in America in 1608
    • Habitat degradation and overharvest lead to population declines
    • Oyster culture seen as solution to saving natural stocks while continuing to meet consumer demand
  • 3. Cultivated Oysters
    • Atlantic oyster ( Crassostrea virginica )
    • Kumamoto oyster ( Crassostrea sikamea )
    • European flat oyster ( Ostrea edulis )
    • Pacific oyster ( Crassostrea gigas )
    • Olympia oyster ( Ostrea conchaphila )
  • 4. Pacific oyster (Crassostrea gigas) :
    • Valuable commercial species
    • Imported from Japan
    • Growth tends to be rapid when they are young and typically decreases when they reach 4-5 years of age.
  • 5. Olympia oyster (Ostrea conchaphila) :
    • Native species
    • This species experiences a slower growth rate, taking 4-5 years to reach market size (50mm).
  • 6. Growth is dependent on …
    • Genetics
    • Environmental factors
      • Water temperature
      • Food availability
      • Placement in the water column
      • Sediment type
      • Density of the bed
  • 7. Objectives
    • Compare growth rates in two species of oysters from the same environment.
    • Identify genes that are likely involved in growth in Pacific and Olympia oysters.
    • Compare gene expression patterns from Pacific and Olympia tissues extracted during two periods of development.
  • 8.
    • Oysters were grown in Agate Pass in Kitsap County, WA.
    • Measurements taken from umbo to edge, once a month from August to December 2007.
    • Tissues samples from the mantle, gills, and muscle were taken in September and November.
    Objective 1: Growth Rates
  • 9.  
  • 10. Objective 2: Gene Identification
    • Expressed sequence tags using bioinformatic techniques (KSPI, PKCIP, INSIG-2, P450)
    • One previously described gene (mGDF)
  • 11. Molluscan growth differential factor (mGDF)
    • associated with the transforming growth factor beta family of proteins.
    • detected in the muscle of O. conchaphila and all tissues from C. gigas .
    m g ma L C. gigas m g ma O. conchaphila L
  • 12. Kazal-type serine peptidase inhibitor domain 1 (KSPI)
    • A Serine proteinase inhibitor found in blood plasma, saliva, secretions of pancreas, seminal vesicles, and submandibular glands.
    • detected in all tissues samples of both O. conchaphila and C . gigas.
    m g ma C. gigas L m g ma O. conchaphila L
  • 13. Protein kinase C inhibitor protein 1 (PKCIP)
    • part of a family of conserved regulator proteins
    • plays regulatory roles in multiple cellular processes including differentiation, cell growth, secretion, and muscle contraction.
    • detected in the gill and muscle of O. conchaphila and all tissues of C. gigas .
    m g ma C. gigas L m g ma O. conchaphila L
  • 14. Insulin-induced gene 2 protein (INSIG-2)
    • involved in metabolic activity, gene transcription, and cell growth.
    • not detected in any tissue of either species.
    m g ma C. gigas L m g ma O. conchaphila L
  • 15. Cytochrome P450 17-hydroxylase/lyase (P450)
    • a monooxygenase enzyme
    • functions in the metabolism of endogenous compounds such as aromatic hydrocarbons
    • In mollusks expression is highest in the digestive gland, but it is also found in blood cells, gills, foot, and gonads.
    • detected in the gill and muscle of C. gigas .
    m g ma C. gigas L m g ma O. conchaphila L
  • 16. Objective 3: Gene Expression
    • Quantitative RT-PCR methods measured and compared gene expression levels in oyster tissue samples.
    x x muscle muscle muscle Olympia Gill X mantle mantle mantle Pacific P450 INSIG2 PKCIP KSPI mGDF Species Tissues Sampled
  • 17.  
  • 18.  
  • 19.  
  • 20.  
  • 21. Conclusion
    • Four genes associated with growth were successfully identified in these oysters.
    • Characterization of tissue expression patterns
    • Quantification of expression level in relation to different periods of development.
    • With the exception of KSPI all the genes examined varied in expression levels over time. This may indicate a switch in metabolic processes from the growing season (summer) to fasting season (winter).
  • 22. Acknowledgements
    • Steven Roberts
    • Sam White
    • Vivianne Barry and Debbie Kay
    • Ken Fulkerson
  • 23. ??Questions??