Expression profiling by high throughput sequencing
Summary
The objective is to elucidate the role of the circadian clock in the parathyroid glands. We investigate the parathyroid transcriptome of wildtype mice (Bmal1 flox/flox) and of mice with parathyroid-specific excision of the circadian clock gene Bmal1 (PTHcre;Bmal1 flox/flox) harvested at 4 hour interval for 24 hours. Rhythmic genes were identified by JTK_CYCLE analysis and revealed 1455 rhythmic genes of the Bmal1 flox/flox and 1055 rhythmic genes of the PTHcre;Bmal1 flox/flox. We found global damepning of circadian clock genes with abolished rhythmicity of Cry1, Cry2, Clock, Npas2, Rorc, and Usp2. We used GSEA analysis to investigate differential expression at each timepount and found downregulation of genes involved in ATP synthesis in PTHcre;Bmal1 flox/flox mice at 4 consecutive timepoints during the active period of the mouse.
Overall design
Samples are pooled parathyroid glands from two mice of either Bmal1 flox/flox (WT) or PTHcre;Bmal1 flox/flox (KO) genotype. Samples were collected at 6 timepoints with 4 hour interval starting when lights turn on at zeitgebertime (=ZT) 0. In total 21 samples from WT and 18 samples from KO were collected.
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