Ai14 mice

Experimental procedures involving live animals were carried out in accordance with animal welfare regulations and with approval of the Regierung Oberbayern (Az 55.2-1-54-2532-187-15 and ROB-55.2–2532.Vet_02-14-157) or in accordance with the Karolinska Institutet’s guidelines for the care and use of animals in research, and were approved by the institute’s Animal Ethics Committee respectively (Ethical permit number 19462–2017). C57Bl/6J B6 albino and C57Bl/6J mice were obtained from The Jackson Laboratory (Maine, USA). Ai14 mice and Cdh5-Cre mice were obtained from the Jackson laboratory (stock number 007914 and 017968). Bbs4^-/- mice were generated in the Lupski lab and backcrossed to C57Bl/6 J (Eichers et al., 2006 (link)). Heterozygous offspring were crossed to produce homozygous Bbs4^-/- and littermate control. Pifo^wt/- mice were crossed to produce homozygous Pifo^-/- mice and littermate controls (Kinzel et al., 2010 (link)). βICKO animals were generated by crossing Pdx1-CreER mice (Zhang et al., 2005 (link)) with Ift88^loxP/loxP mice (Volta et al., 2019 (link); Davenport et al., 2007 (link)).

Animal experiments were approved by the National Institute Health and Harvard Medical School Institutional Animal Care and Use Committee, following ethical guidelines described in the US National Institutes of Health Guide for the Care and Use of Laboratory Animals. For INTACT we crossed Sst-IRES-Cre (The Jackson Laboratory Stock # 013044), Vip-IRES-Cre (The Jackson Laboratory Stock # 010908) and Pv-Cre (The Jackson Laboratory Stock # 017320) with SUN1-2xsfGFP-6xMYC (The Jackson Laboratory Stock # 021039) and used adult (6–12 wk old) male and female F1 progeny. For PESCA screening we used adult (6–10 wk) C57BL/6J (The Jackson Laboratory, Stock # 000664) mice. For confirmation of hits we crossed Sst-IRES-Cre (The Jackson Laboratory Stock # 013044) and Vip-IRES-Cre (The Jackson Laboratory Stock # 031628) mice with Ai14 mice (The Jackson Laboratory Stock # 007914) and used adult (6–12 wk old) male and female F1 progeny. All mice were housed under a standard 12 hr light/dark cycle.

Mice were group housed (two to five animals per cage) in a colony under a 12/12 h light/dark cycle. Food and water were available ad libitum. C57BL/6 (n = 10), DAT-ires-cre (n = 4), DAT-ires-Cre::Ai14 (n = 17), and VGlut2-ires-Cre::Ai14 (n = 9) were used in this study. All mice were two to five months old male. In this study, we focused on the histochemical character of males. DAT-ires-Cre mice (stock #006660; Bäckman et al., 2006 (link)) and Ai14 mice (stock #007908; Madisen et al., 2010 (link)) were purchased from The Jackson Laboratory. VGlut2-ires-Cre mice (Vong et al., 2011 (link)) were provided by Dr. Brad Lowell (Beth Israel Deaconess Medical Center, Harvard Medical School).

Vasoactive intestinal peptide-positive (VIP)-Cre transgenic mice were obtained from Josh Huang’s lab (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, United States). VGAT-Cre mice, Ai14 mice and ChAT-IRES-Cre transgenic mice were obtained from the Jackson Laboratory.
To genetically label specific neurons, we crossed VIP-Cre mice and ChAT-ires-Cre mice with Ai14 reporter mice (VIP-Cre::Ai14 mice and ChAT-IRES-Cre::Ai14 mice), in which the tdTomato was expressed in a Cre-dependent manner. Mice were housed on a 12-h light/dark cycle with food and water ad libitum. Male mice older than 8 weeks were used in the study. All animal experiments followed procedures approved by the Animal Advisory Committee of Huazhong University of Science and Technology.