Cerebellin

Kinase experiment:

Some adrenomedullary samples are incubated with 10 μM H89, U-73122 or calphostin-C alone or in the presence of 0.1 μM cerebellin. When cyclic-AMP production is assayed, 100 μM 3-isobutyl-1-methylxanthine is added to prevent cyclic-AMP metabolism by phosphodiesterases. Incubation is carried out for 60 min (aldosterone or corticosterone production), 30 min (catecholamine production) or 10 min (cyclic-AMP production) in a shaking bath at 37°C in an atmosphere of 95% air -5% CO2. At the end of the experiments, the incubation tubes are centrifuged at 4°C, and media are collected and kept frozen at -80°C[1].

Animal experiment:

Rats[2]Adult female Wistar rats (180–190 g body weight, 8–12 weeks of age) are injected daily with 0.2 mL of 0.9% NaCl (s.c.) for 14 days. On day 15, animals receive s.c. injections of 0.2 mL of 0.9% NaCl (control groups), while experimental groups are treated with 0.5 or 1.5 nmol/100 g body weight Cerebellin0.5 or 1.5 nmol/100 g body weight Cerebellin. Rats are decapitated 60 or 120 min after injection. Blood is collected into ice-cold tubes containing EDTA (1 mg/mL) and analyzed for the concentration of glucose. Plasma is separated and stored at −80 °C for the determination of the insulin and glucagon concentration[2].

References:

[1]. Albertin G, et al. Cerebellin stimulates the secretory activity of the rat adrenal gland: in vitro and in vivo studies. Neuropeptides. 2000 Feb;34(1):7-11.
[2]. Strowski MZ, et al. Insulinostatic activity of cerebellin--evidence from in vivo and in vitro studies in rats. Regul Pept. 2009 Oct 9;157(1-3):19-24.