Asenapine maleate

Melperone hydrochloride (FUJIFILM Wako Pure Chemical Co., Osaka, Japan), ketanserin tartrate (FUJIFILM), methysergide maleate (Sigma-Aldrich, St. Louis, MO, USA), asenapine maleate (Sigma-Aldrich), and SB269970 (Sigma-Aldrich) were added to 10 μM FSW to inhibit the Serotonin/monoamine pathway. The inhibitors were applied 10 s before photoirradiation. S-Nitroso-N-acetyl-D,L-penicillamine (SNAP; FUJIFILM) was used as a nitric oxide (NO) donor (final concentration of 100 μM) [23 ]. SNAP was applied to the larvae 5 min before observation. 3,5-Difluorophenyl-acetyl-L-alanyl-L-S-phenylglycine T-butyl ester (DAPT; Sigma-Aldrich) was used as a γ-secretase inhibitor (final 20 mM). Serotonin (Tokyo Chemical Industry, Tokyo Japan) was dissolved to distilled water just before use and applied to culture (final 10 μM). The same volume of DMSO or seawater was applied as controls for chemical treatments.

The following antibodies were used: anti-AR N-20 (sc-816, Santa Cruz) and anti-β-actin (A5441, Sigma). DHT (5a-Androstain-17b-ol-3-one) and Asenapine maleate were purchase from Sigma. siRNAs specific for TPH1 (TPH1HSS110923) and 5-HT7 (HTR7HSS105140) were purchased from Invitrogen. siRNA for AR (s1539) was purchased from Ambion. A nonspecific control siRNA pool (siControl) was purchased from Dharmacon (D-001206-13-20).