Gabazine sr 95531 hydrobromide

4-AP, TTX, cadmium chloride, and IbTX were purchased from Sigma-Aldrich (St. Louis, MO). Gabazine (SR 95531 hydrobromide) was purchased from Tocris (Bristol, UK). For Per1 gene knock down, either antisense ODN sequence against the 5′ translational start site of Per1 (taggggaccactcatgtct; Sigma-Aldrich) or a random ODN with equivalent GC content (ccgttagtactgagctgac) was applied at a final concentration of 10 µM from ZT 2 to ZT 4. These antisense ODN and random ODN were previously used (Akiyama et al., 1999 (link); Wakamatsu et al., 2001 (link); Tischkau et al., 2003 (link); Marquez et al., 2004 (link); Koyanagi et al., 2006 (link); Gamble et al., 2007 (link); Kudo et al., 2013 (link)). While siRNA can be a more efficient method, there are problems of transfection efficiency while the ODN method to reduce Per1 has been established in the SCN slices (Gamble et al., 2007 (link); Kudo et al., 2013 (link)). The antisense ODN is incorporated into cells by endocytosis. The antisense ODN was applied from ZT 2 to ZT 4, because the SCN slices was moved from 35-mm culture dish to the perfusion chamber. In the perfusion chamber, the flow rate is 2 ml min⁻¹ and it is cost prohibitive to continually apply antisense ODN through the perfusion system.

MPH was purchased from Sigma-Aldrich (St. Louis, MO), and diluted to a 5 mM stock solution in physiological saline. Prepared solution was stored at −20 °C in 1 ml aliquots. Tetrodotoxin, (TTX) was purchased from Abcam (Cambridge, MA) and stored in 1 mM stock at −20 °C. DNQX was purchased from Tocris (Minneapolis, MN), diluted to 20 mM stock and stored at 4 °C. R,S-CPP was purchased from Abcam (Cambridge, MA), diluted to 10 mM stock and stored at −20 °C. AP5 was purchased from Sigma-Aldrich (St. Louis, MO) and was diluted to a 50 mM stock and stored at −20 °C. Gabazine (SR-95531 hydrobromide) was purchased from Tocris (Minneapolis, MN) and was stored at −20 °C in a 2.5 mM stock.