Optogenetic Manipulations of Vglut2+ Neurons

Ten mice were used. We used Vglut2-Cre mice [Jackson laboratories, #028863, Vglut2-ires-cre knock-in (C57BL/6J); Vong et al., 2011 (link); Figures 1A,B], ChR2-EYFP-lox mice (Ai32 mice, Jackson laboratory, #024109, B6.Cg-Gt(ROSA)26Sor^{tm32(CAG-COP4*H134R/EYFP)Hze}/J; Madisen et al., 2012 (link); Figure 1B), and ZsGreen-lox mice (Ai6 mice, Jackson laboratory, #007906, B6.Cg-Gt(ROSA)26Sor^{tm6(CAG-ZsGreen1)Hze}/J; Madisen et al., 2012 (link); Figure 1A). We crossed homozygous Vglut2-Cre mice with homozygous ChR2-EYFP-lox mice to obtain the double heterozygous Vglut2-ChR2-EYFP mice. We crossed homozygous Vglut2-Cre mice with homozygous ZsGreen-lox mice to obtain the double heterozygous Vglut2-ZsGreen mice. Animals had ad libitum access to food and water, with lights on from 6 AM to 8 PM. Mice were 16–36 weeks old for in vivo optogenetics (three males, two females), 10–18 weeks old for neuroanatomy (one male, two females), and 15–23 days old for patch-clamp experiments (one male, one undetermined).