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. 2013 Dec 13;21(4):568–581. doi: 10.1038/cdd.2013.177

Figure 6.

Figure 6

Trpv5/6 has different roles in regulating IGF-PI3K-Akt signaling under normal and low [Ca2+]. (ad) Effects of cadmium (Cd2+), ruthenium red (RR), lanthanum (La3+), and gadolinium (Gd3+) on pAkt. Larvae (72 hpf) were transferred to 0.2 mM or 0.001 mM [Ca2+] water containing the indicated concentrations of the drugs. After 8 h, they were stained for pAkt and scored. The total number of larvae analyzed from three independent experiments is shown on the top of each column. (e) Blockage of the Trpv5/6 channel abolishes the low [Ca2+]-induced increase in NaR cell density. Larvae (72 hpf) were transferred to 0.2 mM or 0.001 mM [Ca2+] water containing the indicated concentrations of Cd2+. At 120 hpf, they were sampled and analyzed by in situ hybridization for igfbp5a. (f) The low [Ca2+]-induced Akt signaling is not mediated by intracellular Ca2+/CaM signaling. Larvae (72 hpf) were transferred to 0.001 mM [Ca2+] water containing BAPTA-AM (100 μM), W7 (50 μM), or Calmidazolium (Calm, 1 μM). After 8 h, they were fixed, stained for pAkt, and scored as described earlier. (g) KCl treatment activates Akt signaling in an IGF1R and PI3K-dependent manner. Larvae (72 hpf) were transferred to 0.2 mM or 0.001 mM [Ca2+] water containing KCl (100 mM), sorbitol (200 mM), NMDG-Cl (100 mM), BMS (0.3 μM), and Wortmannin (0.06 μM), alone or in combination. After 2 h, they were stained for pAkt. (h) KCl reverses the effect of Trpv5/6 blockage. Larvae (72 hpf) were transferred to 0.2 mM or 0.001 mM [Ca2+] water containing KCl (100 mM), RR (5 μM), and Cd2+ (1 μM), alone or in combination. After 2 h, they were stained for pAkt