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Proteome Profiling of Developing Murine Lens Through Mass Spectrometry
Shahid Y Khan1, Muhammad Ali1, Firoz Kabir1
1The Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States.
Investigative Ophthalmology & Visual Science
|January 15, 2018
Summary
Researchers profiled the mouse lens proteome using mass spectrometry. They identified thousands of proteins, revealing key developmental changes and proteins linked to lens transparency and diseases like cataracts.
Area of Science:
- Ophthalmology
- Proteomics
- Developmental Biology
Background:
- Previous research established a comprehensive mouse lens transcriptome profile.
- Understanding the lens proteome is crucial for maintaining ocular transparency.
Purpose of the Study:
- To investigate the mouse lens proteome using mass spectrometry-based protein sequencing.
- To analyze protein expression at various embryonic and postnatal developmental stages.
Main Methods:
- Mouse lenses were collected at embryonic day 15 (E15) and 18 (E18), and postnatal days 0 (P0), 3 (P3), 6 (P6), and 9 (P9).
- Proteins were extracted, digested, labeled with isobaric tandem mass tags (TMT), and analyzed across three independent TMT experiments.
- Biological replicates were maintained for each developmental stage.
Main Results:
- A total of 5404 proteins were identified in the mouse ocular lens.
- 3155 proteins were consistently detected across all three TMT experiments.
- 39 proteins showed an 8-fold differential expression during development, with significant proteins linked to cataractogenesis and autophagy.
Conclusions:
- A comprehensive profile of the developing murine lens proteome has been established.
- This proteomic data serves as a valuable resource for identifying critical components in lens development.
- The findings aid in understanding processes essential for maintaining lens transparency.
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