Vacutainer plus blood collection tubes

To determine if the animals were responsive to Kisspeptin following treatment, experiment 7 included two treatment groups: Control (n = 4) and SC300 (n = 3). On PND 60–70 (2–2.5 months) after treatment on PND 0.5, peripheral blood (50 µL from the ventral tail vain) was drawn and serum collected and immediately afterwards the females were injected intraperitoneally with a Kisspeptin-10 analog (KP-10; 50 nmole, Millipore-Sigma, San Diego, CA). After 30 min, blood was drawn again and serum collected. Serum LH concentration was measured by a sandwich immunoassay using monoclonal antibodies against both bovine LH (Cat. 581B7, Medix Biochemica, Kauniainen, Finland) and human LH-beta subunit (Cat. 5303, Medix Biochemica), at the University of Virginia Center for Research in Reproduction Ligand Assay and Analysis Core, as previously described (range 0.04–37.4 ng/mL)^{33 (link)}. Blood cell population in control and treated rats were analyzed by collecting 1 mL whole blood by cardiac puncture using BD Vacutainer^® Plus blood collection tubes coated with a spray-dried K2 EDTA (Becton Dickinson, Franklin Lakes, NJ, USA).

Experiment 3 included four treatment groups: Control (n = 8), EBx11 (n = 6), SC300 (n = 5), SC30 (n = 9). Experiment 4 included two treatment groups: Control (n = 6) and SC300 (n = 5), For some endpoints, data were combined from both experiments: body and organ weights, serum chemistry, CBC and hormone measurements. On PND 178–211 (6–7 months), body weights were determined and the stage of estrous cycle and vaginal opening were evaluated for each animal. Blood was then collected for serum chemistry, CBC and hormone measurements (described below) and the females were then euthanized. The anovaginal distance (AVD) was measured and the major organs removed and weighed. The ovaries, the uteri and livers were collected for histological evaluation and mammary gland tissues were collected for whole mount analysis (described below). Blood cell population (CBC) was analyzed from 1 mL whole blood that was collected by cardiac puncture using BD Vacutainer^® Plus blood collection tubes coated with a spray-dried K2 EDTA (Becton Dickinson, Franklin Lakes, NJ, USA). Immediately after collection, samples were gently inverted several times and complete blood count (CBC) analyses were conducted at the University of Illinois Veterinary Medicine Diagnostics Laboratory (Urbana, IL).