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Related Concept Videos

Circadian Rhythms and Gene Regulation02:19

Circadian Rhythms and Gene Regulation

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The biological clock is involved in many aspects of regulating complex physiology in all animals. It was in 1935 when German zoologists, Hans Kalmus and Erwin Bünning, discovered the existence of circadian rhythm in Drosophila melanogaster. However, the internal molecular mechanisms behind the circadian clock remained a mystery until 1984, when Jeffrey C. Hall, Michael Rosbash, and Michael W. Young discovered the expression of the Per gene oscillating over a 24-hour cycle. In subsequent...
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Circadian Gene Profiling in Laser Capture Microdissected Mouse Club Cells.

Zhenguang Zhang1, Andrew Loudon1

  • 1School of Medical Science, Faculty of Biology, medicine and Health, University of Manchester, Manchester, UK.

Bio-Protocol
|March 4, 2021
PubMed
Summary
This summary is machine-generated.

This study details a reproducible RNA sequencing protocol for analyzing the circadian clock in pure mouse lung club cells. This method provides deeper insights than traditional bulk tissue analysis.

Keywords:
Circadian clockClub cellLaser capture micro-dissectionLungRNA seq

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Area of Science:

  • Cellular Biology
  • Genomics
  • Chronobiology

Background:

  • Tissues like the lung exhibit significant cellular heterogeneity.
  • Studying pure cell populations offers greater insights than bulk tissue analysis, particularly for complex biological processes like the circadian clock.
  • Understanding cellular heterogeneity is crucial for accurate biological research.

Purpose of the Study:

  • To provide a detailed protocol for circadian clock studies in specific lung cell types.
  • To enable RNA sequencing analysis on laser capture micro-dissected mouse lung club cells.
  • To establish a reproducible method for investigating circadian rhythms in isolated cells.

Main Methods:

  • Utilized laser capture micro-dissection to isolate specific mouse lung club cells.
  • Employed RNA sequencing (RNA seq) for gene expression analysis.
  • Developed a workflow optimized for frozen tissue samples.

Main Results:

  • Successfully established a detailed workflow for circadian clock studies.
  • Demonstrated the feasibility of applying RNA sequencing to micro-dissected lung cells.
  • The protocol was found to be highly reproducible.

Conclusions:

  • This protocol enables precise circadian clock analysis in isolated mouse lung club cells.
  • The method overcomes limitations of bulk tissue analysis by focusing on pure cell populations.
  • The high reproducibility ensures reliable data for future chronobiology research.