using the zebrafish to understand and develop …using the zebrafish to understand and develop...
TRANSCRIPT
Usingthezebrafishtounderstandanddeveloptreatmentsforcomplexbraindisorders
Camila V. Esguerra, PhD Group Leader, Chemical Neuroscience Group Centre for Molecular Medicine Norway Assoc. Professor, Department of Pharmacy University of Oslo [email protected]
SLC6A1 Round Table American Epilepsy Society Meeting
New Orleans, USA 29 November 2018
CONFIDENTIAL
Chemical Neuroscience Group
What is our research about? • Study the causes of brain disorders
– Epilepsy, Schizophrenia/Bipolar, Autism • Create new animal models of human disease • Use models to screen for novel drugs • Understand how these drugs work • Test drug candidates for potential toxicities
What is our long-term goal? • Develop new, safe and effective medicines
C.Esguerra,SLC6A1RoundTable,AES2018
Zebrafish: a novel way of "fishing"
• Vertebrate model with fully sequenced, annotated genome • Strong genetic, physiological and pharmacological similarity to humans • High fecundity and small size (96-well format) • Rapid development ex utero • Optical transparency (non-invasive imaging) • Only µg amounts of compounds needed; readily absorbed (skin, GI tract, gills)
C.Esguerra,SLC6A1RoundTable,AES2018
C.Esguerra,SLC6A1RoundTable,AES2018
Chemical Neuroscience Group Platform Mutants
EEGconfirma/onofseizureinhibi/on
Compoundlibraries
Pharmacology,ToxicologyHistology,RNA-Seq,ImagingNeurotransmiHerProfiling
wild-type mutant
Behavioralassays
invivodrugscreen
PhenotypicAnalysis
Gene/czebrafishmutantforsevereearlyonsetepilepsysyndrome
PaEents:• Physicallyunderdeveloped• Slowmovements• Seizures• Au/s/cfeaturesZebrafishmodel:• Physicallyunderdeveloped• Slowtouchresponse• Seizures• Au/s/cfeatures
5dayspostfer/liza/onwildtype(A)andmutant(B)
C.Esguerra,SLC6A1RoundTable,AES2018
C.Esguerra,SLC6A1RoundTable,AES2018
Behaviorallarvallocomotorassay(automatedvideotracking)
C.Esguerra,SLC6A1RoundTable,AES2018
Alteredshoalingbehavioringene/czebrafishmutant
P<0.0087 7
A B wt
s a 1 5 2 9 6
s a 1 0 9 3 0 0
5
1 0
1 5
N N D
G e n o ty p e
Ne
are
st
ne
igh
bo
ur
dis
tan
ce
, m
m **
n s
Wild type Mutant 1 Mutant 2
(A) Wild type
(C) Wild type + Drug (D) Mutant + Drug
Neuronalbranchingdefectsingene/cmutant
(B) Mutant
wt
s c n 1 lab- /-
wt +
FE N
s c n 1 lab- /- + F
E N
0
2 0 0
4 0 0
6 0 0
Arb
ori
zati
on
(A
U) ****
a
Wildtype Mutant Mutant+drug
Wildtype+drug
Chronicdrugtreatment
C.Esguerra,SLC6A1RoundTable,AES2018
GABA
ergicneuron
s
wt
s c n 1 lab- /-
0
2 0 0
4 0 0
6 0 0
Arb
ori
zati
on
(A
U)
*Wild type Mutant
NeuronalstructuraldefectsprecedeseizuresGA
BAergicneuron
s
Wild type Mutant
C.Esguerra,SLC6A1RoundTable,AES2018
C.Esguerra,SLC6A1RoundTable,AES2018
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−40
−20
0
20
40
−25 0 25Component 1
Com
pone
nt 2
Cluster
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●1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
ASTROCYTES
MICROGLIA
NEURONS
OPC
RADIALGLIA
ERYTHROCYTES
EPITHELIALCELLS
Incollabora/onwithA.Skupin,LuxembourgCenterforSystemsBiomedicine
Geneexpressionandcellclusterprofiling
WT day 4 scn1lab −/− day 4 WT day 7 scn1lab −/− day7
% cell type
Samples0
2040
6080
100
Astrocytes (CLS #1 #9 #12)Microglia (CL #11)Neurons (CLS #3 #5)OPC (CL #10)Radial glia (CL #6)Erythrocytes (CL #4)Epithelial Cells ( CL #7)
Wildtypeearlyseizure
onset
Mutantearlyseizure
onset
Wildtyperecurringseizures
Mutantrecurringseizures
Epilepsy models
Antiseizure hits from medicinal plants