Sf1 cre

Mice expressing Cre recombinase under the control of the steroidogenic factor 1 promoter (Sf1-Cre; stock #012462), and mice expressing the cre-dependent reporter, tdTomato (Ai14, stock #007914), were obtained from The Jackson Laboratory. Similarly, mice containing alleles with LoxP sites that flank the coding exon of the Bdnf gene (Bdnf^flox/flox; stock# 004339) were also obtained from The Jackson Laboratory. The Sf1-Cre mouse line was on a FVB/NJ background, the tdTomato mouse line was on a C57/BL6J background, and the Bdnf^flox/flox mouse line was on a 129S4/SvJae background. Therefore, progeny produced by intercross of these lines to produce Sf1-Cre; tdTomato;Bdnf^flox/flox mice, and littermates, were maintained on a mixed background. The terms Sf1-Cre; tdTomato;Bdnf^+/+ or control mice refer to mice with normal expression of the Bdnf gene, while the term Sf1-Cre; tdTomato;Bdnf^flox/flox mice refers to mice with targeted deletion of the Bdnf gene in SF1 neurons. All animals were housed at 22 °C on a 13/11-hour light/dark cycle (lights on at 0600 h/lights off at 1900 h) with ad libitum access to a standard chow diet (PicoLab Rodent Diet 20, #5053). All animal care and experimental procedures were performed in accordance with the Institutional Animal Care and Usage Committee of the Saban Research Institute, Children's Hospital Los Angeles.

To specifically disrupt the BBSome in SF1 neurons, mice bearing the Bbs1 gene flanked by loxP sites (Bbs1^fl/fl), on C57BL6 background, were bred with mice expressing Cre recombinase driven by the SF1 promoter (SF1^Cre, Jackson Laboratory, stock#012462), on a mixed background (129 SvEv, FVB/NJ and C57BL/6 J). To validate the specificity of Cre-dependent activity to the VMH, we used a td-Tomato reporter mouse on a mixed background (129SvEv and C57BL/6 J). This mouse has a stop codon flanked by loxP sites preceding the start position of a td-Tomato locus (Stop^fl/fl-tdTomato). Cre recombination removes the stop site, leading to td-Tomato fluorescence protein expression that can be visualized under a fluorescence microscope. A male SF1^cre mouse was bred with a female Stop^fl/fl-tdTomato mouse. Genotyping of the mice was determined using standard polymerase chain reaction (PCR). Tail DNA was extracted by using ethanol precipitation, and PCR amplification was performed using primers provided in Table S1. PCR product was assayed in ethidium bromide gel (2% agarose in Tris-acetate-EDTA).
All animal testing was approved by the University of Iowa Animal Care and Use Committee. Mice were maintained on a 12:12-hour light–dark cycle (light off at 6 PM) in temperature (23 °C)-controlled rooms with access to mouse chow (Teklad 7913) and tap water ad libitum.