Twelve zebra finches were separated into three rearing conditions 4-5 days after hatching. Four zebra finches with an equal distribution in gender were randomly assigned to the three different rearing conditions; biparentally reared, maternally deprived, or paternally deprived. For the following 115 days, the interactions between parental and offspring behaviour was recorded. On PHD 30, 90, and 120, baseline blood samples were taken within five minutes of entering the laboratory room. Corticosterone levels from the blood samples were collected for the experiment. However, this report is focusing on GCR levels, and not including Corticosterone levels in the blood. On PHD 120, the acute stressor occurred. The stressor consisted of 30 minutes of restraint in a cloth bag. Immediately preceding the stressor on PHD 120, the 12 zebra finches were sacrificed and their brains were sectioned to be stained for GCRs. …show more content…
The slides were prepared using the Antibody Immunohistochemistry procedure for GCR protein (PA1510A Life Technologies at 1:1000) that used DAB to stain the collected tissue to locate GCR proteins. Primary antibody is created so there is a high affinity for the protein in the experiment. For this study, the primary antibody is selective for GCR protein. The secondary antibody has a high affinity for the first, and is also bound to HRP. In the presence of GRCs, the primary antibody will bind to the GRCs. The secondary antibody will bind to the primary and then DAB will fluoresce brown in the presence of HRP. This is due to the oxidation of