FIGURE

Fig. 3

ID

ZDB-FIG-180913-48

Publication

Ashlin et al., 2018 - Pitpnc1a Regulates Zebrafish Sleep and Wake Behavior through Modulation of Insulin-like Growth Factor Signaling

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Fig. 3

pitpnc1a^?/? Brains Have Increased Neuronal Activity

(A?B?) ISH reveals many areas of c-fos expression in 6 dpf pitpnc1a^?/? larval brains (A and A?) compared with wild-type (B and B?).

(C?D?) Following a 1 hr light pulse, additional neurons express c-fos in pitpnc1a^?/? (C and C?) and wild-type (D and D?). Many areas expressing c-fos in wild-type after a light pulse overlap with strong c-fos signals in pitpnc1a^?/? brains at 0 hr (black arrowheads, D and D?).

(E?F?) After 2 hr of light, c-fos levels remain elevated in pitpnc1a^?/? brains (E and E?) but have returned to baseline in wild-type (F and F?).

(G) pERK/tERK comparison of dark-reared, 6 dpf pitpnc1a^?/? (n = 15) and wild-type (n = 12) brains reveals mutant brain areas with upregulated (green) and downregulated (magenta) expression of pERK relative to controls. White arrowheads indicate areas of agreement between pERK and c-fos expression, and white arrows show the downregulated signal. Data are shown as a thresholded maximum projection overlaid on a maximum projection of the Z-Brain tERK reference (gray).

(H) Clustering the behavioral fingerprint of pitpnc1a^?/? against 550 drug-induced behavioral profiles places the phenotype in a strong day-active and modestly night-active cluster (red branches).

(I) Flumethasone increases the behavioral activity of both wild-type and pitpnc1a^?/? to the same level. DMSO-treated pitpnc1a^?/? animals are hyperactive (^?p < 0.05, one-way ANOVA, Tukey?s post hoc test). Both 10 and 30 ?M flumethasone increase larval activity (^??p < 0.01, two-way ANOVA, gene × dose, Tukey?s post hoc test).

(J) Flumethasone induces c-fos expression in areas similar to pitpnc1a^?/? and light pulses (black arrows).

Hb, hindbrain; POA, preoptic area. (A?F) Dorsal view. (A??F?) Lateral view. (J) Ventral view. Scale bars, 100 ?m.

Expression Data

Gene:	fosab
Antibodies:	Ab9-mapk Ab12-mapk
Fish:	AB/TL pitpnc1a^u506/u506 (AB/TL)
Conditions:	chemical treatment by environment: flumethasone constant dark constant light altered light dark cycle
Anatomical Terms:	brain cerebellum diencephalon dopaminergic neuron forebrain dorsal region forebrain ventral region hindbrain hypothalamus hypocretin-secreting neuron hypothalamus posterior region locus coeruleus midbrain optic tectum preoptic area superior raphe nucleus (all 13)
Stage:	Day 6

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	AB/TL pitpnc1a^u506/u506 (AB/TL)
Conditions:	chemical treatment by environment: flumethasone constant dark constant light altered light dark cycle
Observed In:	cerebellum circadian sleep/wake cycle, wakefulness diencephalon dopaminergic neuron forebrain dorsal region forebrain ventral region hindbrain hypothalamus hypocretin-secreting neuron hypothalamus posterior region locus coeruleus midbrain musculoskeletal movement optic tectum preoptic area superior raphe nucleus (all 14)
Stage:	Day 6

Phenotype Detail

Acknowledgments

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