K2e edta vacuette tube

After finding that GRA treatment significantly impacted conditioned avoidance behavior and cFos activation (see Results), we sought to confirm our assumption that plasma acyl-ghrelin levels increase in rats exposed to single and repeated sessions of overnight food deprivation. For this, terminal blood samples were collected from a separate cohort of adult male rats that were either ad lib fed with no fasting experience (n=3), fasted once overnight before morning blood collection (n=3), or fasted overnight on 3 separate occasions (with at least two days between each episode) with the 3rd fast occurring before morning blood collection (n=4). Prior to blood collection, rats were injected with pentobarbital sodium (39 mg/ml i.p., Fatal Plus Solution; Butler Schein), and blood samples were collected within 5 min from the right atrium of the heart using a 3mL syringe pre-rinsed with 1% EDTA. Blood samples were transferred to a K2E EDTA vacuette tube (BD Ref 367841), and aprotinin (50µl/mL) was added to each sample. Samples were centrifuged at 3500 rpm for 10 min at 4°C to separate the plasma. Hydrochloride was added to the supernatant (10µl/100mL) centrifuging again at 3500 rpm for 5 min at 4°C. Plasma samples were stored at −80°C. Acyl-ghrelin was measured using an ELISA kit (BioVendor, R&D; RA394062400R) according to the manufacturer’s instructions.

After finding that GRA treatment significantly impacted conditioned avoidance behavior and cFos activation (see Results), we sought to confirm our assumption that plasma acyl-ghrelin levels increase in rats exposed to single and repeated sessions of overnight food deprivation. For this, terminal blood samples were collected from a separate cohort of adult male rats that were either ad lib fed with no fasting experience (n=3), fasted once overnight before morning blood collection (n=3), or fasted overnight on 3 separate occasions (with at least two days between each episode) with the 3rd fast occurring before morning blood collection (n=4). Prior to blood collection, rats were injected with pentobarbital sodium (39 mg/ml i.p., Fatal Plus Solution; Butler Schein), and blood samples were collected within 5 min from the right atrium of the heart using a 3mL syringe pre-rinsed with 1% EDTA. Blood samples were transferred to a K2E EDTA vacuette tube (BD Ref 367841), and aprotinin (50µl/mL) was added to each sample. Samples were centrifuged at 3500 rpm for 10 min at 4°C to separate the plasma. Hydrochloride was added to the supernatant (10µl/100mL) centrifuging again at 3500 rpm for 5 min at 4°C. Plasma samples were stored at −80°C. Acyl-ghrelin was measured using an ELISA kit (BioVendor, R&D; RA394062400R) according to the manufacturer’s instructions.