Cell Type-Specific Functions of Period Genes Revealed by Novel Adipocyte and Hepatocyte Circadian Clock Models

• DOI: 10.1371/journal.pgen.1004244
• Publication Date: 2014-04-03T16:29:58Z
• Authors: Chidambaram Raman...; Haiyan Xu; Sanjoy K. Khan; Yang Shen; Paula J. Gitis; David K. Welsh; John B. Hogenesch; Andrew C. Liu
• Keywords: PER2; PER1; Cell type

In animals, circadian rhythms in physiology and behavior result from coherent rhythmic interactions between clocks the brain those throughout body. Despite many tissue specific clocks, most understanding of molecular core clock mechanism comes studies suprachiasmatic nuclei (SCN) hypothalamus a few other cell types. Here we report establishment genetic characterization three cell-autonomous mouse models: 3T3 fibroblasts, 3T3-L1 adipocytes, MMH-D3 hepatocytes. Each model is genetically tractable has an integrated luciferase reporter that allows for longitudinal luminescence recording gene expression using inexpensive off-the-shelf microplate reader. To test these cellular models, generated library short hairpin RNAs (shRNAs) against panel known genes evaluated their impact on rhythms. Knockdown Bmal1, Clock, Cry1, Cry2 each resulted similar phenotypes all consistent with previous studies. However, observed type-specific knockdown Period Rev-Erb families genes. particular, Per1 Per2, which have strong behavioral effects knockout mice, appear to play different roles regulating period length amplitude peripheral systems. Per3, relatively modest substantially affects models dissociated SCN neurons. summary, this study establishes new are particular relevance metabolism suitable screening modifiers, reveals previously under-appreciated functions