Using the zebrafish to understand and develop …Using the zebrafish to understand and develop...
Transcript of Using the zebrafish to understand and develop …Using the zebrafish to understand and develop...
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