Generation of Tanycyte-Specific LepR-KO Mice

Rax-CreER^T2 mice generated in the laboratory (Pak et al., 2014 (link)) (JAX#025521) were bred with LepR^lox/lox mice (Cohen et al., 2001 (link)) (JAX #008327) to generate tanycyte-specific LepR-KO mice. Rax-CreER^T2;Ai9 (R26-CAG-lsl-tdTom, JAX #007909) and Rax-CreER^T2;CAG-Sun1/sfGFP (Mo et al., 2015 (link)) (JAX #021039) were bred in the laboratory. To induce Cre recombination, tamoxifen was administered by either i.p. injection (1 mg, Sigma-Aldrich #H6278) at P28 for 3 consecutive days for fluorescent reporter expression, or by feeding commercial tamoxifen-containing diet (EnvigoTeklad diets #TD.130856) for 3 weeks to delete LepR from tanycytes. Rax-EGFP BAC transgenic line (MMRRC #030564-UCD) was originally generated by the Gene Expression Nervous System Atlas Brain Atlas (GENSAT) Project (Gong et al., 2003 (link)). 7 weeks old C57BL/6 male mice were purchased from the Charles River Laboratories and used for scRNA-Seq analysis. All mice were housed in a climate-controlled pathogen free facility on a 14 h-10 h light/dark cycle (07:00 lights on – 19:00 lights off). All experimental procedures were pre-approved by the Institutional Animal Care and Use Committee (IACUC) of the Johns Hopkins University School of Medicine.