Dat ires cre

The following mouse lines (25-30 g, 8-12 weeks old, male) were used for the experiments: C57Bl6 mice (Jackson Laboratory), DAT::IRES-Cre (Jackson Laboratory, stock number: 006660, strain code: B6.SJL-Slc6a3tm1.1(cre)Bkmn/J), VGLUT2::IRES-Cre (Jackson Laboratory, stock number: 016963, strain code: Slc17a6tm2(cre)Lowl/J), VGLUT3::IRES-Cre (Jackson Laboratory, stock number: 018147, strain code: Tg(Slc17a8-icre)1Edw/SealJ), GAD2::IRES-Cre (Jackson Laboratory, stock number: 010802, strain code: Gad2^tm2(cre)Zjh/J), Ai14 Cre reporter mice (Jackson Laboratory, stock number: 007908, strain code: B6;129S6 Gt(ROSA)26Sor^{tm14(CAG-tdTomato)Hze}/J). Ai14 Cre reporter mice were crossed to GAD2::IRES-Cre and VGLUT2::IRES-Cre mice. Mice were maintained on a 12:12 light cycle (lights on at 07:00). All procedures complied with the animal care standards set forth by the National Institutes of Health and were approved by University of California Berkeley’s Administrative Panel on Laboratory Animal Care.

Methods were adapted as previously described (Stamatakis et al., 2013 (link)). Briefly, Vgat-ires-Cre: (Slc32a1tm2(cre)Lowl/J, Jackson Laboratory stock number: 016962, Dat-ires-Cre: (B6.SJL-Slc6a3tm1.1(cre)Bkmn/J, Jackson Laboratory stock number: 006660, and TH-ires-Cre: (B6.129X1-Thtm1(cre)Te/Kieg, European Mouse Mutant Archive stock number: EM:00254) were used. Mice were injected bilaterally with AAV-DIO-ChR2-eYFP, (AAV5 ~10¹² infectious units/mL, packaged and titered by the UNC Vector Core Facility).

The following mouse lines (25–30 g, 8–12 weeks old, male) were used for the experiments: C57BL/6J mice (Jackson Laboratory), DAT-IRES-Cre (Jackson Laboratory, stock number: 006660, strain code: B6.SJL-Slc6a3^{tm1.1(cre)Bkmn}/J), VGLUT2-IRES-Cre (Jackson Laboratory, stock number: 016963, strain code: Slc17a6^tm2(cre)Lowl/J), GAD2-IRES-Cre (Jackson Laboratory, stock number: 010802, strain code: Gad2^tm2(cre)Zjh/J), VGAT-IRES-Cre (Jackson Laboratory, stock number:028862, strain code: B6J.129S6(FVB)-Slc32a1^tm2(cre)Lowl/MwarJ), Ai14 Cre reporter mice (Jackson Laboratory, stock number: 007908, strain code: B6;129S6 Gt(ROSA)26Sor^{tm14(CAG-tdTomato)Hze}/J). GAD2-Cre-tdTomato: Ai14 Cre reporter mice were crossed to GAD2-IRES-Cre mice. For ex vivo patch-clamp recordings shown in Figure 1g–m and Figure 2a–d male and female mice were used, but no significant differences were observed between male and female mice. Mice were maintained on a 12:12 light cycle (lights on at 07:00) with water and food ad libitum and room temperature of 22–25°C and 55% humidity. All procedures complied with the animal care standards set forth by the National Institutes of Health and were approved by University of California Berkeley’s Administrative Panel on Laboratory Animal Care.

A total of 247 adult male mice were used. Strain details and number of animals in each group are as follows:
183 C57BL/6J (Jackson Laboratories stock #000664)
9 VGat-ires-Cre (Slc32a1^tm2(cre)Lowl/J (Jackson Laboratories stock #016962)
40 VGlut2-ires-Cre (Slc17a6^tm2(cre)Lowl/J (Jackson Laboratories stock #016963)
2 Dat-ires-Cre (Slc6a3^{tm1.1(cre)Bkmn}/J (Jackson Laboratories stock #006660)
2 Dat-ires-Cre × Ai14 = Dat-ires-Cre (Slc6a3tm1.1(cre)Bkmn/J (Jackson Laboratories stock #006660) × Ai14 (B6.Cg-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J (Jackson Laboratories stock #007914)
1 Agrp-ires-Cre × Ai34D = Agrp-Cre (Agrpt^m1(cre)Low (Jackson Laboratories stock #012899) × Ai34D (B6;129S-Gt(ROSA)26Sor^{tm34.1(CAG-Syp/tdTomato)Hze}/J (Jackson Laboratories stock #012570) 10 Ai14 (B6.Cg-Gt(ROSA)26Sor^{tm14(CAG-tdTomato)Hze}/J (Jackson Laboratories stock #007914)
All mice used in experiments were individually housed under a 12 hr light/dark cycle. At the time of the experiments, animals were 8–20 weeks old. Animals weighted approximately 25–28 grams. All animals were used in scientific experiments for the first time. This includes no previous exposures to pharmacological substances or altered diets. Health status was normal for all animals.

A total of 59 mice (51 male; 8 female) were used in this study, in which 49 mice (44 male; 5 female) and 10 mice (7 male; 3 female) were used in validating the HSV and AAV CRISPR vectors, respectively. Animal study protocols pertaining to this study (#IS00003282 and #IS00003604) were reviewed and approved by the Northwestern University Institutional Animal Care and Use Committee. Procedures also followed the guidelines for the care and use of animals provided by the National Institutes of Health Office of Laboratory Animal Welfare. Mice were housed at 22 °C on a 12-hour light/dark cycle with food and water ad libitum. Mice were weaned on postnatal day 21 and housed with same-sex littermates. The following mouse strains were obtained from Jackson Laboratories: VGLUT2-IRES-Cre (Jax #016963), DAT-IRES-Cre (Jax #06660), Rosa26-LSL-Cas9 (Jax #026175), C57BL/6 (Jax #000664). Mice expressing Cas9-2A-EGFP in a Cre-dependent manner (VGLUT2-Cre::Cas9, DAT-Cre::Cas9) were obtained by crossing mice heterozygous for each mutation. All reasonable efforts were made to minimize pain and discomfort. These efforts include, but were not limited to, use of anesthetic during surgery (see below for dosage), post-operative care pain management (5 mg/kg ketoprofen, subcutaneous (s.c.)), and reduction of animal use by collecting multiple independent data samples from the same mouse.

All procedures were approved by the IACUC of NYULMC in compliance with the NIH guidelines for the care and use of laboratory animals. Mice were housed at 18–23°C with 40–60% humidity under a 12 h light–dark cycle (dark cycle, 10 p.m. to 10 a.m.), with food and water available ad libitum. Test animals were adult Drd1-cre (>8 weeks, RRID: MMRRC_030989-UCD) and DAT-ires-cre (>8 weeks, Jackson, RRID: IMSR_JAX:006660) mice. Ai9 mice (Jackson, RRID: IMSR_JAX:007909) were crossed with Drd1-cre mice for revealing D1R expression. Stimulus animals were adult C57BL/6N male and female mice, adult BALB/c male and female mice, or sexually experienced C57BL/6N and Swiss Webster males (aggressor) purchased from Charles River, and 3–7 days old pups were from test mice or wildtype C57BL/6N breeders. After surgery, all the animals are single housed. All experiments were performed during the dark cycle of the animals.