Sprague dawley rats

All experimental protocols were approved by the Experimental Animal Ethics Committee at Asahi Kasei Pharma Corp (Tokyo, Japan) and conducted in accordance with established guidelines concerning the management and handling of experimental animals. Three-month-old female Sprague-Dawley rats (Charles River, Yokohama, Japan) were housed in a dedicated laboratory animal facility with a 12-h light/dark cycle and unrestricted access to tap water and food (CRF-1; standard diet of rats; Oriental Yeast, Tokyo, Japan).
At six months old, the rats were randomly assigned to one of the following body weight-matched groups: sham ovariectomy (Sham) group (n = 8), ovariectomy (OVX) group (n = 8) or OVX-teriparatide administration (OVX-TPTD) group (n = 8); after assignment, they underwent bilateral ovariectomy or sham ovariectomy as appropriate. Two months after the operation, saline or 6.0 μg/kg teriparatide was subcutaneously injected 3 times/week for 4 months. Serum samples were obtained by centrifugation of blood samples collected from the subclavian vein in the morning on the last administration day. The CCL5 levels in the rat serum were measured using a Mouse/Rat CCL5/RANTES Quantikine ELISA Kit (R&D Systems, Minneapolis, MN, USA).

A total of forty-two 9-week-old male Sprague-Dawley rats weighting 234 ± 23 g were purchased from Oriental Yeast Co. (Tokyo, Japan) and housed in a controlled environment at 22°C on a 12-hour light, 12-hour dark cycle. Animal experiments were approved by Committee of Animal Experiment at Kobe University School of Medicine and conducted in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals [19 ].

Pregnant Sprague-Dawley rats (Oriental Yeast Co., Osaka, Japan) were gavage-fed with 100 mg of the herbicide nitrofen (2,4-dichloro-4-nitrofenil ether; Tokyo Chemical Industry, Tokyo, Japan) dissolved in 1.0 mL of olive oil at embryonic day (E) 9.5 (predicted term: E22, n = 12) as previously described [9 (link)], whereas control rats received olive oil only (n = 4). The dose and administration timing of ONO-1301SR were determined based on the results of a preliminary study. Of the 12 nitrofen-administered rats, 5 received a single subcutaneous injection of 30 mg/kg ONO-1301SR (Ono Pharmaceutical Co. Ltd., Osaka, Japan) dissolved in saline at E9.5. The following 3 groups were thus generated: control (n = 4), nitrofen administration (n = 7), and nitrofen+ONO-1301SR administration (n = 5). At term (E21.5), the pregnant rats were anesthetized and the fetuses were harvested by cesarean section. Fetuses were weighed and examined for diaphragmatic defects, and the lungs were harvested for further pathobiological evaluation. The fetuses that had macroscopically developed diaphragmatic defects in the nitrofen and nitrofen+ONO-1301SR groups were diagnosed with CDH and were further analyzed.