Development and Characterization of the cellular circadian clock of C2C12 cells

Author

Ali Akbari beni

Date

2023

Document Type

Thesis

Degree Name

Master of Science

Department

Nutrition

Committee Chair

Marie Van Der Merwe

Committee Member

Chidambaram Dr. Ramanathan

Committee Member

Marie Dr. Van Der Merwe

Committee Member

Yufeng Dr. Zhang

Abstract

Abstract Circadian rhythms in mammalian physiology and behavior are generated through coherent rhythmic crosstalk between the central clock in the brain and peripheral clocks throughout the organs in our body. The molecular clocks responsible for these intrinsic rhythmic oscillations work based on interlocked transcription/translation feedback loops, which work together with various environmental and metabolic cues to produce internal 24-hour timing. This project established a stable cellular circadian clock model using the C2C12 cell line. This genetically tractable model contains an integrated luciferase reporter, which enables continuous luminescence recording of each circadian cycle. In order to examine this cellular circadian model, I created a panel of short hairpin RNAs (shRNAs) targeting ten of the primary known clock genes (Bmal1, Clock, Per1, Per2, Per3, Cry1, Cry2, Nr1d1, Nr1d2, Fbxl3). I evaluated their functional role in circadian rhythm generation. Knockdown of Bmal1 resulted in arrhythmicity. Knockdown of the Clock caused a shorter period length and a lower amplitude. Knockdown of Per1 resulted in shorter period lengths and Knockdown of Per2 resulted in shorter periods and high amplitude. Per3 knockdown caused a high amplitude phenotype. Knockdown of Cry2 caused short period length and higher amplitude. On the other hand, the knockdown of Cry1 led to lower amplitude. Knockdown of Nr1d1, and Fbxl3 both resulted in lower amplitude with no changes in period length. In summary, I established a cellular clock model of the C2C12 cell line, which is amenable to genetic manipulation and would be ideal for studying the crosstalk between clock and muscle physiology and metabolism.

Comments

Data is provided by the student.

Library Comment

Dissertation or thesis originally submitted to ProQuest

Notes

Open Access

Recommended Citation

Akbari beni, Ali, "Development and Characterization of the cellular circadian clock of C2C12 cells" (2023). Electronic Theses and Dissertations. 3063.
https://digitalcommons.memphis.edu/etd/3063