Single-nucleus RNA sequencing of the mouse hypothalamus in the fed and fasted state

To provide a summary of cells within the mouse hypothalamus in the fed and fasted state, we performed single nucleus RNA sequencing on 12 mice fed ad libitum, and 6 mice that were fasted overnight. Hypothalami were dissected and stored at -80ºC overnight. The next day single nucleus supensions were made, pooling together hypothalamic from the same feeding condition (2 x ad libitum prepared on separate days; 1 x overnight fast). Susepensions were FACS sorted for DRAQ5 positive events. Single nucleus RNA sequencing libraries were generated using the 10X Chromium single cell 3' reagents. Overall design: The hypothalamus was dissected from 12 adlib fed (2 cohorts) and 6 overnight (16 hr) fasted mice, and a single nucleus suspension was created using a dounce homogenizer and purified using FACS, sorting for DRAQ5 positive events. These samples (2 adlib fed and 1 overnight fasted) were submitted for single nucleus RNA sequencing