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. 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为总结自由进食与禁食状态下小鼠下丘脑内的细胞特征，我们对12只自由进食（ad libitum）的小鼠以及6只隔夜禁食的小鼠开展单细胞核RNA测序（single nucleus RNA sequencing）。首先获取小鼠下丘脑组织并于-80℃条件下隔夜保存，次日制备单细胞核悬液：将同一饲喂条件下的下丘脑组织混合混匀，其中2批自由进食样本分别于不同日期制备，1批隔夜禁食样本。随后通过荧光激活细胞分选（Fluorescence Activated Cell Sorting，FACS）筛选DRAQ5阳性事件，并使用10X Chromium单细胞3'端试剂构建单细胞核RNA测序文库。本次实验共从12只自由进食（分为2个队列）以及6只隔夜（16小时）禁食的小鼠体内获取下丘脑组织，采用杜恩匀浆器（dounce homogenizer）制备单细胞核悬液，通过FACS筛选DRAQ5阳性事件完成纯化。上述2份自由进食样本与1份隔夜禁食样本均提交进行单细胞核RNA测序。