ZFIN Image: Ashlin et al., 2018, Fig. 4

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Figure Caption

Fig. 4

IGFR Signaling Is Enhanced in pitpnc1a^?/? Brains, and Inhibiting IGFR Signaling Rescues Neuronal and Behavioral Hyperactivity

(A?E) 6 dpf pitpnc1a^?/? larvae have increased pIGFR puncta in the brain. (A?B?) Confocal projections of the dorsal telencephalon in wild-type (A and A?) and pitpnc1a^?/? (B and B?) reveal pIGFR puncta (green). Nuclei are DAPI stained (blue) in (A?) and (B?). Confocal projections of the pitpnc1a^?/? ventral hypothalamus (D and D?) have more pIGFR puncta (C and C?), as quantified in (E) (mean � SEM, one-way ANOVA).

(F) Upregulated c-fos in pitpnc1a^?/? embryos is reduced to wild-type by soaking in 1 ng/?L IGFBP2 from 36 to 48 hpf.

(G?J) ISH for c-fos in 6 dpf brains in constant dark after overnight exposure to PI3K (LY-294002, 2 ?M) or Akt (MK-2206, 100 nM) inhibitors. (G and H) c-fos is upregulated in pitpnc1a^?/? brains (G) relative to WT controls (H). (I and J) Akt (I) and PI3K (J) inhibitors reduce mutant c-fos expression to wild-type levels. Brains in (G)?(J) are overdeveloped to expose even weak c-fos expression.

(K) Exposure to 1 ?M Akt inhibitor, MK-2206, reduces average night-time activity of 6 dpf pitpnc1a^?/? larvae to wild-type levels. Each dot represents a single larva, and the crossbars plot the mean � SEM (two-way ANOVA, genotype � drug interaction, Tukey?s post hoc test).

(L) Model of Pitpnc1a modulation of neuronal activity via IGFBP2 and inhibition of IGF-PI3K-Akt. Loss of Pitpnc1a enhances IGF signaling, leading to increased waking behavior.

(G?J) Ventral view; scale bars, 100 ?m. (A?B?) Scale bars, 100 ?m. (C?E?) Scale bars, 20 ?m.

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Acknowledgments

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