Adult mice (8–12 week-old female mice or 14-16 week-old male mice) were bilaterally ovariectomized or castrated. For sex hormone replacement, a 2 cm length of silastic tubing (inner diameter: 1.58 mm; outer diameter: 3.18 mm) containing 36 μg 17β-estradiol/mL (E2758, Sigma)42 in sesame oil or testosterone powder43 (link) (T1500, Sigma) were inserted on the dorsal aspect of the animal’s neck immediately after gonadectomy. Mice were allowed to recover for 1 week before further treatment.
For ablation experiments, 8- to 10-week-old female mice or 14- to 16-week-old male mice were intraperitoneally injected with 20 ng/g bodyweight diphtheria toxin (DT; 322326, EMD Millipore) twice with a 3-day interval between injections. For activation experiments, 2.5 mg of clozapine-N-oxide (CNO; Tocris Bioscience) in 200 ml of drinking water was administered to the mice with a high-fat diet (22% carb, 24% protein, 54% fat, E15742-347, Ssniff). For FSH administration, mice with a high-fat diet received daily 30 IU/kg i.p FSH (GONAL-f, Merck) injections. Mice were weighed and monitored daily before being euthanized as experimental mice. Mice were humanely euthanized if postsurgical complications progressed to a pre-defined humane endpoint at which the mice started to suffer.
For ablation experiments, 8- to 10-week-old female mice or 14- to 16-week-old male mice were intraperitoneally injected with 20 ng/g bodyweight diphtheria toxin (DT; 322326, EMD Millipore) twice with a 3-day interval between injections. For activation experiments, 2.5 mg of clozapine-N-oxide (CNO; Tocris Bioscience) in 200 ml of drinking water was administered to the mice with a high-fat diet (22% carb, 24% protein, 54% fat, E15742-347, Ssniff). For FSH administration, mice with a high-fat diet received daily 30 IU/kg i.p FSH (GONAL-f, Merck) injections. Mice were weighed and monitored daily before being euthanized as experimental mice. Mice were humanely euthanized if postsurgical complications progressed to a pre-defined humane endpoint at which the mice started to suffer.
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