ZFIN ID: ZDB-FIG-180710-53
Marsden et al., 2018 - A Cyfip2-Dependent Excitatory Interneuron Pathway Establishes the Innate Startle Threshold. Cell Reports   23:878-887 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Anatomical Terms:
Stage: Day 5
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage: Day 5

Fig. 2

Hypersensitivity of triggerhappyp400 Mutants Caused by cyfip2 Mutations and Rescued by Conditional Cyfip2-GFP Expression

(A) Acoustic startle circuit. Acoustic nerve (VIII), posterior lateral line nerve (PLL), feedforward (FF) inhibitory, and excitatory spiral fiber (SF) neurons connect to the Mauthner cells (red).

(B) Cyfip2 protein interaction domains (Abekhoukh and Bardoni, 2014, Pittman et al., 2010). triggerhappyp400(cyfip2p400) mutants have a premature stop codon after 342 of 1,253 amino acids. The previously identified nevermind (cyfip2tr230b) mutation (Pittman et al., 2010) is shown.

(C) Startle sensitivity curves of siblings and trans-heterozygous (trans-het) larvae from cyfip2p400/+ X cyfip2tr230b/+ crosses (n = 75 siblings, 34 trans-hets; mean ± SEM).

(D) Startle sensitivity index in cyfip2p400 sibling and mutant larvae expressing Tg(hsp70:cyfip2-GFP). Larvae were given no heat shock or one 40-min heat shock at 30 hpf. Cyfip2-GFP fluorescence was largely restricted to the CNS and was visible 90 min after heat shock, peaked around 3 hr after heat shock, and was detectable at low levels 24 hr later (Figure S3C). Without a heat shock, cyfip2p400 mutants had increased startle sensitivity (∗∗∗p < 0.001, Mann-Whitney test), whereas heat shock reduced the sensitivity of cyfip2 mutants with the transgene compared with those without it (∗∗p < 0.01, Mann-Whitney test).

(E) Startle sensitivity curves for fmr1 sibling (n = 62) and mutant larvae (fmr1hu2787/hu2787, n = 20) at 5 dpf (mean ± SEM).

(F) Hindbrain expression of Cyfip2 in 5 dpf wild-type (cyfip2+/+) and mutant (cyfip2p400/p400) larvae using a Cyfip2 antibody (Ab). Membranes of VIII neurons are marked by Tg(SCP1:Gal4FF(y256Et));Tg(UAS:gap43-citrine) and anti-GFP Ab. Dashed lines indicate the otic vesicles (OVs). Scale bar, 10 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
Citrine pglyrp5y256Et ; cyfip2p400/p400 ; p201Tg standard conditions Day 5 statoacoustic (VIII) ganglion neuron IHC
Day 5 statoacoustic (VIII) ganglion plasma membrane IHC
pglyrp5y256Et ; p201Tg standard conditions Day 5 statoacoustic (VIII) ganglion neuron IHC
Day 5 statoacoustic (VIII) ganglion plasma membrane IHC
cyfip2 Ab1-cyfip2 pglyrp5y256Et ; cyfip2p400/p400 ; p201Tg standard conditions Day 5 hindbrain IHC
Ab1-cyfip2 pglyrp5y256Et ; p201Tg standard conditions Day 5 hindbrain IHC
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT standard conditions Day 5 startle response occurrence, normal
Day 5 startle response sensitivity of a process sensory perception of sound, normal
WT heat shock Day 5 startle response occurrence, normal
Day 5 startle response sensitivity of a process sensory perception of sound, normal
cyfip2p400/+ standard conditions Day 5 startle response occurrence, normal
Day 5 startle response sensitivity of a process sensory perception of sound, normal
cyfip2p400/+ heat shock Day 5 startle response occurrence, normal
Day 5 startle response sensitivity of a process sensory perception of sound, normal
cyfip2p400/+ ; cyfip2tr230b/+ standard conditions Day 5 startle response increased occurrence, abnormal
Day 5 startle response increased sensitivity of a process sensory perception of sound, abnormal
cyfip2p400/p400 standard conditions Day 5 startle response increased occurrence, abnormal
Day 5 startle response increased sensitivity of a process sensory perception of sound, abnormal
cyfip2p400/p400 heat shock Day 5 startle response increased occurrence, abnormal
Day 5 startle response increased sensitivity of a process sensory perception of sound, abnormal
fmr1hu2787/+ standard conditions Day 5 startle response occurrence, normal
Day 5 startle response sensitivity of a process sensory perception of sound, normal
fmr1hu2787/hu2787 standard conditions Day 5 startle response occurrence, normal
Day 5 startle response sensitivity of a process sensory perception of sound, normal
p407Tg ; cyfip2p400/+ standard conditions Day 5 startle response occurrence, normal
Day 5 startle response sensitivity of a process sensory perception of sound, normal
p407Tg ; cyfip2p400/+ heat shock Day 5 startle response occurrence, normal
Day 5 startle response sensitivity of a process sensory perception of sound, normal
p407Tg ; cyfip2p400/p400 standard conditions Day 5 startle response increased occurrence, abnormal
Day 5 startle response increased sensitivity of a process sensory perception of sound, abnormal
p407Tg ; cyfip2p400/p400 heat shock Day 5 startle response occurrence, ameliorated
Day 5 startle response sensitivity of a process sensory perception of sound, ameliorated
pglyrp5y256Et ; cyfip2p400/p400 ; p201Tg standard conditions Day 5 hindbrain cyfip2 expression decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Rep.