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Exit International’s founder Dr Philip Nitschke, known as Dr Death, warned some of the sites are scams. The resulting RT-PCR fragments were subcloned into the HindIII-XbaI sites of pBluescript II (SK-) (Agilent Technologies, Santa Clara, CA, USA). Quantitative RT-PCR (qRT-PCR) was performed as described previously24,44,45,46,47. To detect dnBMAL1 mRNA and arginine vasopressin (Avp) mRNA, double-label fluorescence in situ hybridization (FISH) analysis was performed according to a previously described protocol43,44,49,50,51 with a few modifications. After quenching with H2O2, FISH signal of the digoxigenin-labeled Avp riboprobe was detected with anti-digoxigenin-POD, Fab fragments (Roche) and amplified with a cyanine-5 substrate kit (Cy5 TSA Plus, PerkinElmer). FISH signal of the fluorescein-labeled dnBMAL1-specific riboprobe was detected with anti-fluorescein horseradish peroxidase (HRP) conjugate (PerkinElmer, Boston, MA, USA) and amplified with a cyanine-3 substrate kit (Cy3 TSA Plus; PerkinElmer). To generate a specific riboprobe for TRE promoter-dependent transcripts encoding dnBMAL1, the 0.3-kbp HindIII-XbaI fragment containing the 3′-UTR from pTRE-dnBMAL1 was cloned into pBS (pBS-TRE 3′-UTR). To generate a riboprobe for Avp mRNA, Avp cDNA (cds, 21-495) was amplified by RT-PCR using mouse brain cDNA as a template (and with the following primers: Avp forward, GGGAAGCTTCACTACGCTCTCCGCTTGTT; Avp reverse, GGGTCTAGAGGGCTTGGCAGAATCCA). The NotI fragment encoding dnBMAL1 was subcloned into the NotI site of pcisTREmlu43, generating pTRE-dnBMAL1. The dnBMAL1 transgene contains the 3′-end of BMAL1 open reading frame and the 3′-untranslated region (3′-UTR) from pTRE-dnBMAL1 that produces dnBMAL1 mRNA-specific PCR products. Total RNA was prepared from mouse whole-hippocampus region using the RNeasy Mini Kit (Qiagen, Valencia, CA, USA) or acid guanidinium thiocyanate-phenol-chloroform extraction method43,44,45,46,47,48. Specific probes for southern blotting of TRE-dnBMAL1 mice and CaMKII-tTA mice were derived from the 0.5-kbp NotI-NdeI fragment containing the TRE-promoter region from pcisTRE43 and the 0.6-kbp EcoRI-HindIII fragment encoding TetR region from pcDNA3-TetR-KRAB, respectively43,44,45,46,47,48. 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