Using high throughput sequencing method such as single-cell RNA-Sequencing (scRNA-Seq) method combined with behavioural functions in adult (sleep and motivation), we plan to understand how the hypothalamus is formed during development and how various cell types from individual hypothalamic regions can have distinct physiological functions.

With development, we are conducting large-scale scRNA-Seq analysis to understand how the hypothalamus is patterned by various transcription factors (TFs) during development in mice (hypothalamic patterning) and how TF mutants can result in defect(s) in hypothalamic patterning (mutant screening). Without our extensive understanding in mouse hypothalamus development, we plan to understand hypothalamic development of other species such as chicken, zebrafish and human (cross-species comparison).

With physiological functions focusing on sleep and motivation, we are trying to identify changes in molecular expression across different physiological conditions (i.e. sleep and wake) and use this information to identify new cell type(s). With intersectional approaches (Flp and Cre recombination) combined with optogenetic and/or pharmacogenetic approaches, we are planning to test physiological functions of the newly identified cell types.

12 timepoints of developing diencephalon including prethalamus and hypothalamus (between E10 and E16), and hypothalamus (between E18 and P45) were profiled using 10x Genomics Chromium system. 2D tSNE showing distribution of individual ages.
3D tSNE plot showing divisions of developing diencephalon (prethalamus and hypothalamus) and nearby regions to the developing diencephalon.

Distribution of individual ages (see first figure) is shown in 3D UMAP plot.

Dong Won Kim, Parris Whitney Washington, Zoe Qianyi Wang, Sonia Lin, Changyu Sun, Lizhi Jiang, Seth Blackshaw. Single cell RNA-Seq analysis identifies molecular mechanisms controlling hypothalamic patterning and differentiation. Science, submitted.

https://doi.org/10.1101/657148