Isolation of Oxytocin Receptor-Rich Nuclei from a Neonatal Rat Brain

The brain's oxytocin receptor-rich nuclei are clusters of neurons that express oxytocin receptors, which interact with oxytocin, a neuropeptide.

To isolate oxytocin receptor-rich nuclei, begin with a neonatal rat brain.

Place the brain in a mold with a slicing guide. Slice the entire brain to expose different regions with nuclei.

Transfer these slices to a dish and immobilize them.

Add artificial cerebrospinal fluid, or aCSF, to maintain neuronal viability and incubate under gentle rotation.

Using a published brain atlas as a reference, identify anatomical landmarks to locate oxytocin-receptor-rich nuclei within the slices.

Transfer an identified brain slice of interest to a new plate.

Under a microscope, locate the nuclei within the slice and punch it.

Repeat the process for other identified sections to isolate the oxytocin-rich nuclei.

Transfer the isolated nuclei to a tube containing ice-cold protein extraction buffer supplemented with protease and phosphatase inhibitors to preserve the oxytocin receptors.

After harvesting the brain, rapidly place the whole organ in a polymethylmethacrylate brain mold at room temperature, and immediately use a new razor blade to make 500-microme