Dissection of hypothalamus

The brain was quickly removed, shock frozen on dry ice, and stored at −80 °C. For further processing, the brains were trimmed in a cryochamber of −17 °C with a razor blade. The hypothalamus was clearly visible as a central elevation on the ventral side of the brain expanding from Bregma 0.38 mm to −2.80 mm (according to the mouse brain atlas [21]). The hypothalamus was first trimmed to a tissue cuboid cutting from ventral to dorsal. To remove the thalamus and the cortex, brains were cut from anterior to posterior directly on the anterior part of the anterior commissure, visible as most ventral bilateral white matter. Edges of the tissue cuboid were removed cutting from ventral to dorsal to resemble the cylindric shape of the hypothalamus.