Fig. S4
Related to Figure 4 and Figure 6: PI3K/mTOR inhibition rescues dendritic melanophores in bace2-/-. (A) Representative pictures of melanophores with chemical screen scores from 0 (non-rescued, mutant like) to 5 (fully rescued, WT like). (B) Confocal pictures high magnification view of tailfin melanophores in response to PI3K inhibitor AS605240 (110nM), mTOR inhibitor PP242 (15μM) and insulin receptor inhibitor NVP-AEW541 (60μM). Tg(tyrp1b: membrane- mCherry) fish were treated from 24hpf to 72hpf to visualize melanophore membrane structure. Also see Supplemental Movie 1-4. (C) PI3K/mTOR/Insulin receptor inhibitors effect on tail melanophore phenotype is not reversible by drug washout. wanderlust mutants were treated with inhibitors from 24hpf to 72hpf, then inhibitors were washed off extensively and embryos were imaged at 108hpf. (D-E) bace2-/- is hypersensitive to PI3K/mTOR inhibition compared to WT fish. PI3K/mTOR inhibition reduced WT melanophore dendricity at 48hpf (D, arrowhead), but the bace2-/- melanophores are more sensitive to this effect at both 48hpf and 72hpf (D and E, arrow). Treatment of WT and bace2-/- embryos with the PI3K inhibitor AS605240 (110nM) or mTOR inhibitor Temsirolimus (30μM) from 24-48hpf (D) or from 24-72hpf (E). Scale bars, 30μM (B), 100μM (A, C, D, E). |
Reprinted from Developmental Cell, 45(5), Zhang, Y.M., Zimmer, M.A., Guardia, T., Callahan, S.J., Mondal, C., Di Martino, J., Takagi, T., Fennell, M., Garippa, R., Campbell, N.R., Bravo-Cordero, J.J., White, R.M., Distant Insulin Signaling Regulates Vertebrate Pigmentation through the Sheddase Bace2, 580-594.e7, Copyright (2018) with permission from Elsevier. Full text @ Dev. Cell