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Epigenetic and Environmental
Influences on the Shellfish
Immune Response
Steven B. Roberts
Assistant Professor
School of Aquatic and Fishery Sciences
University of Washington
Seattle - Washington - US
Open Science
•You are free to Share!
•Our lab practices open notebook
science
•Slides, links, and related material are
available @
oystergen.es/vigo
Photo credit: Flickr, Creative Commons, dougbrown47
Immune Function
e
Immune Function
e
e
Epigenetics
Immune Function
e
Immune Function
Temperature
Genetics
Reproductive Status
Resources
Culture Practices
Salinity
Pathogen
e
Immune Function
e
Characterization of genes involved in ceramide metabolism in the Pacific oyster
(Crassostrea gigas).
Timmins-Schiffman and Roberts 2012
Immune Function
e
Rapid accumulation of an interleukin 17 homolog transcript in Crassostrea gigas
hemocytes following bacterial exposure.
Roberts et al 2008
Analysis of genes isolated from plated hemocytes of the Pacific Oyster,
Crassostrea gigas.
Roberts et al 2009
Characterization of genes involved in ceramide metabolism in the Pacific oyster
(Crassostrea gigas).
Timmins-Schiffman and Roberts 2012
e
O c e a n A c i d i f i c a t i o n
e
Characterizing short read sequencing for gene discovery and
RNA-Seq analysis in Crassostrea gigas.
Gavery and Roberts 2012
Elevated pCO2 causes developmental delay in early larval Pacific oysters, Crassostrea gigas.
Timmins-Schiffman et al 2012
O c e a n A c i d i f i c a t i o n
e
Proteom
e
Immune Function
O c e a n A c i d i f i c a t i o n
e
O c e a n A c i d i f i c a t i o n
unpublished
Proteomic Response
e
O c e a n A c i d i f i c a t i o n
unpublished
Proteomic Response
oxidation reduction
signal transduction
oxidative stress
response
cellular stress &
transport
Elevated (7.2) Control (8)
e
Immune Function
e
Immune Function
e
Immune Function
e
unpublished
Control
e
unpublished
Puta%ve	
  cytochrome	
  P450
Isocitrate	
  dehydrogenase	
  [NAD]	
  subunit	
  beta
Alpha-­‐soluble	
  NSF	
  aCachment	
  protein
C16orf62	
  homolog
Radial	
  spoke	
  head	
  protein	
  4	
  homolog	
  A
Flo%llin-­‐2
Small	
  nuclear	
  ribonucleoprotein-­‐associated	
  protein
ES1	
  protein	
  homolog
Sterile	
  alpha	
  and	
  TIR	
  mo%f-­‐containing	
  protein	
  1
Twinfilin-­‐2
Lachesin
Syntaxin-­‐5
Serine	
  dehydratase-­‐like
Caspase-­‐3
tRNA	
  methyltransferase	
  112	
  homolog
Circularly	
  permutated	
  Ras	
  protein	
  1
Contac%n
Caspase-­‐7
Apopto%c	
  chroma%n	
  condensa%on	
  inducer
Calcium-­‐binding	
  protein
Vacuolar	
  protein	
  sor%ng-­‐associated	
  protein	
  13C
Cullin-­‐1
Control
e
unpublished
O c e a n A c i d i f i c a t i o n
Control Elevated
e
Immune Function
Epigenetics
Photo credit: Flickr, Creative Commons, he-boden
Epigenetics
Epigenetics
Histone Modification
Epigenetics
short RNAs
DNA Methylation
Histone Modification
Epigenetics
short RNAs
DNA Methylation
m
m
m
m
DNA Methylation
Function?
e
Epigenetics
Transcriptom
e
traditional - vertebrate model
Nature Reviews Genetics 9, 465-476 (June 2008)
methylation
Non-
Vertebrates?
Absent in
several
model
organisms
Function?Shellfish?
mosaic
associated with gene bodies
Photo credit: Flickr, Creative Commons, dkeats
oyster
mosaic
associated with gene bodies
oyster
mosaic
associated with gene bodies
methylationmosaic
Roberts and Gavery 2012
in silico analysis
Roberts and Gavery 2012
in silico analysis
Roberts and Gavery 2012
in silico analysis
relationship
with
function
associated with gene bodies
Role?
Methylation
Enhanced
Random
Variation
theory
Biology
Biology
Biology
In species that experience a diverse range of
environmental conditions, processes have evolved to
increase the number of potential phenotypes in a
population in order to improve the chances for an
individual’s survival.
M M M M M M M M M M
germline methylation
conventional transcription
5’ UTR promotor
ubiquitously expressed,
critical genes
Transcriptional opportunities
x x
M M M M
sparse methylation
alternative start sites
sequence mutation
change AA, premature stop codon
conventional transcription
transient methylation
alternate transcript
exon skipping
tissue / temporal specific and
inducible genes
Transcriptional opportunities
Roberts and Gavery 2012
DNA methylation in Crassostrea gigas
Gill
Sperm
DNA methylation in Crassostrea gigas
Gill
Sperm
If true?
Limb
Photo credit: Flickr, Creative Commons, csessums
Could change how we consider
tolerance
Could change how we consider
natural variation
Could change how we consider
adaptation
defective
methylation
defective
methylation
Profiling the DNA methylomes of plants exposed
to bacterial pathogen, avirulent bacteria, or salicylic acid
hormone revealed numerous stress-induced
differentially methylated regions, many of which
were intimately associated with differentially
expressed genes.
ChIP-seq analysis of four
histone modifications
revealed that the priming
treatment altered the
epigenomic landscape; ....
preferentially targeted
transcription factors.
Open Science
Open Science
web-native scholarship
Photo credit: Flickr, Creative Commons, speechless
Sharing
How?
Why?
Example
Example
Example
Example
Example
unpublished
Example
unpublished
Open Notebook Science
... there is a URL to a laboratory notebook that is
freely available and indexed on common search
engines. It does not necessarily have to look like a
paper notebook but it is essential that all of the
information available to the researchers to make
their conclusions is equally available to the rest of
the world.
—Jean-Claude Bradley
Open Notebook Science
Open Notebook Science
Open Notebook Science
Open Notebook Science
Open Notebook Science
Hello,
I was doing a search for what
concentrations of MgSO4 to use as a
bivalve relaxant and came across
Katie’s Notebook. What a cool idea!
Please say thanks to Katie :). My boss
said MgSO4 but most of the literature
was MgCl2.... So g’day from the
University of Queensland in Brisbane,
Australia.
Cheers
KA
email June 22, 2013
R e p r o d u c i b i l i t y
usable data formats
Sharing
R e p r o d u c i b i l i t y
usable data formats
Sharing
Platforms
FigShare
Dryad
GitHub
runmycode
R e p r o d u c i b i l i t y
usable data formats
Sharing
looking for patterns
Querying Disparate Datasets
Genome
Primary
Data Table
Groupings
Expressed Sequence Tags
Gene Expression GeneticVariation Epigenetic Features
transcripts
dynamic
RNA-Sequencing Single Nucleotide Polymorphisms
Simple Sequence Repeats
DNA Methylation
Histone Modification
miRNA ExpressionExpression Microarrays
Amplified Fragment
Length Polymorphisms
Genomic
DataTypes
Data Tables
static
Gene Annotations Sequence Motifs
Gene OntologiesPathways
Orthologs
Transposable Elements
Interactions
Other species genomes
CpG statistics
Structural Elements
Size
Growth
Location
Environment
Stage
Treatment
Tissue
Trait
Strain
Transcription Factors
Binding Sites
Publications
Immune Function
e
Immune Function
e
I n s i g h t
i n t o
t h e
C o m p l e x i t y
Acknowledgements
Emma Timmins-Schiffman
Mackenzie Gavery
Claire Ellis
Sam White
Brent Vadopalas
Bill Howe
Dan Halperin
EPA
STAR
Aquaculture Program
Saltonstall-Kennedy
acidification
DNA methylation
Acknowledgements
Emma Timmins-Schiffman
Mackenzie Gavery
Claire Ellis
Sam White
Brent Vadopalas
Bill Howe
Dan Halperin
EPA
STAR
Aquaculture Program
Saltonstall-Kennedy
acidification
DNA methylation
oystergen.es/vigo
Epigenetic and Environmental Influences on the Shellfish Immune Response

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