Hpth 1 34

The rats in the Sham and MM + veh control groups (groups 1 and 2) received vehicle (sterile water used for injections) at 1 mL/kg subcutaneously (sc) 5 days/week, starting on the day of surgery. The rats in group 3 received zoledronic acid (Zol; Sargent Pharmaceuticals 25021-801, Schaumburg, IL, USA) at 100 μg/kg, sc, twice/week [18 (link)], and the rats in group 4 received human PTH (hPTH 1-34; Sigma-Aldrich P3796, St. Louis, MO, USA) at 40 μg/kg, sc, five times/week [25 (link)], starting on the day of surgery. To label actively mineralized bone surfaces, all of the rats in the study received calcein (Sigma-Aldrich Cat. No. C-0875, St. Louis, MO, USA) at 10 mg/kg (3.3 mL/kg) 13 days before necropsy and alizarin (Sigma-Aldrich Cat. No. A-5533, St. Louis, MO, USA) at 30 mg/kg (3.0 mL/kg) 3 days before necropsy.

All hormone injections were performed in the home cage of the mice after random treatment allocation of individual ear‐marked mice. For 1,25(OH)₂‐vitamin D₃ treatment, male C57BL/6N mice aged 13–15 weeks were subcutaneously injected with 2 μg/kg body weight 1,25(OH)₂‐vitamin D₃ (Sigma D1530) in ethanol 1% in NaCl 0.9%. The dose and application route was the same as previously used in our laboratory. Control mice were injected with 1% (v/v) ethanol in NaCl 0.9%. Mice were sacrificed 6 hr after injection.
For PTH treatment, male C57BL/6N mice aged 12–13 weeks were subcutaneously injected with 80 μg/kg body weight human PTH fragment 1–34 (hPTH1‐34) (Sigma P3796) in NaCl 0.9% or NaCl 0.9% alone as vehicle and sacrificed 2 hr after injection. The dose and application route used was determined from the literature (Kramer, Loots, Studer, Keller, & Kneissel, 2010).
For estradiol treatment, male C57BL/6N mice aged 16 weeks received one daily subcutaneous injection of 15 μg 17β‐Estradiol (Sigma E8875) in ethanol 0.1% (v/v) in NaCl 0.9% for five consecutive days and were sacrificed 4 hr after the last injection. The dose per body weight and the drug application route was derived from the literature, as shown Van Abel et al. (2002) to induce Trpv5 expression. Control mice were subcutaneously injected with 0.1% ethanol in NaCl 0.9% for 5 days.