Effects of acute kidney dysfunction on hypothalamic arginine vasopressin synthesis in transgenic rats

Oxytocin‐monomeric red fluorescent protein 1 synthesis in the hypothalamus under osmotic challenge and acute hypovolemia in a transgenic rat line

“…To measure the changes in OXT-mRFP1, transgenic rats were sacrificed by transcardial perfusion at 0, 3, 6, 12, and 24 hr after administration of saline, HTN, or PEG at 10:00-12:00 (n = 6 in each group). All rats were deeply sedated after administration of the anesthetic mixture, which consisted of medetomidine (3 mg/100 g), midazolam (4 mg/100 g), and butorphanol (5 mg/100 kg), before perfusion (Kawai et al, 2011;Ueno et al, 2019). Transcardial perfusion was performed as described previously (Ueno et al, 2018(Ueno et al, , 2019.…”

“…All rats were deeply sedated after administration of the anesthetic mixture, which consisted of medetomidine (3 mg/100 g), midazolam (4 mg/100 g), and butorphanol (5 mg/100 kg), before perfusion (Kawai et al, 2011;Ueno et al, 2019). Transcardial perfusion was performed as described previously (Ueno et al, 2018(Ueno et al, , 2019. After thoracotomy, the heart was carefully exposed.…”

“…A fluorescent protein was used to label c-Fos protein using the immunohistochemical staining method described in previous reports (Motojima et al, 2016;Ueno et al, 2018Ueno et al, , 2019. The second group of sections mentioned above was incubated for 5 days at 4°C with a primary anti-Fos antibody solution (#sc-52; rabbit, Santa Cruz Biotechnology, Dallas, TX, USA; 1:500 in phosphate-buffered saline [PBS]).…”

“…To measure the changes of mRFP1 and OXT gene expressions in the PVN and SON for analysis by in situ hybridization histochemistry, transgenic rats were sacrificed by decapitation at 3 and 6 hr after administration of saline, HTN, and PEG at 10:00-12:00 (n = 6 in each group). In situ hybridization in this study was performed based on previous reports (Ueno et al, 2018(Ueno et al, , 2019Ueta, Levy, et al, 1995). Thus, only a brief explanation of the procedure of in situ hybridization is provided in this article.…”

“…The 35 S-labeled oligodeoxynucleotide probe and protocol for in situ hybridization histochemistry (ISH) in this experiment have been used many times before, and its reliability is considered guaranteed. The details of the probe sequence and the protocol for ISH have been described previously (Ueno et al, 2018(Ueno et al, , 2019Ueta, Levy, et al, 1995). The hybridized sections were exposed to autoradiography films (Amersham Hyperfilm, Buckinghamshire, UK) for 3 days (mRFP1) and 6 hr (OXT).…”