Single nucleus sequecing data of MCH neurons [scRNA-seq]

In order to understand heterogeneity of MCH neurons, by using MCH-Cre dependent ZsGreen (fl/fl) reporter mice, we isolated nuclear from 16 hypothalami of 16 mice at the age of 16-18weeks. Through flow cytometry, we were able to distinguish ZsGreen positive nuclei, and collected puried MCH nuclei in suspension. Those nuclei suspension were subjected to single cell sequence by 10x™ GemCode™ Technology for further single nuleus mRNA analysis and unravel subclusters of MCH neurons Overall design: 16 genotype with MCH-Cre;ZsGreen positive mice with mixed gender at the age of 16-18weeks mice were subjected to acute decapitation. Hypothalami from each mouse were dissected by using brain matrix with blades. Collected hypothalami from each brain were fast frozen in liquid nitrogen and subjected by homogenetization and single nucleus extration. Fresh single nuclei suspension were subjected to flow cytometry and isolated ZsGreen positive nuclei were collected and polled followed by single nucleus sequecing by 10x™ GemCode™ Technology. Further analysis allowed us to get subcluster of MCH neurons from 1266 cells in the end.