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Laser Capture Microdissection of Highly Pure Trabecular Meshwork from Mouse Eyes for Gene Expression Analysis
Published on: June 3, 2018
Serial analysis of gene expression (SAGE) in normal human trabecular meshwork
Yutao Liu1, Drew Munro, David Layfield
1Center for Human Genetics, Duke University Medical Center, Durham, NC, USA.
Molecular Vision
|April 30, 2011
Summary
This study used serial analysis of gene expression (SAGE) to identify genes in normal human trabecular meshwork (HTM). The findings provide insights into the gene expression profile of HTM, crucial for understanding glaucoma pathogenesis.
Area of Science:
- Ophthalmology
- Genomics
- Molecular Biology
Background:
- The trabecular meshwork (TM) is vital for regulating intraocular pressure and is implicated in glaucoma pathogenesis.
- Understanding the gene expression profile of normal human trabecular meshwork (HTM) is essential for elucidating glaucoma mechanisms.
Purpose of the Study:
- To identify genes expressed in normal human trabecular meshwork (HTM) using SAGE.
- To establish a gene expression profile of HTM for glaucoma research.
Main Methods:
- Serial analysis of gene expression (SAGE) was performed on RNA from three human trabecular meshwork donors.
- SAGE libraries were created, and sequence tags were extracted and mapped to the human genome.
- SAGE 2000 software and SAGE Genie map were utilized for data analysis.
Main Results:
- Over 298,000 SAGE tags were identified, with over 10,000 unique tags mapping to known Unigene clusters.
- Approximately 29% of tags did not map to known Unigene clusters, and 13% mapped to multiple clusters.
- Tags corresponding to known glaucoma-related genes, such as myocilin and optineurin, were detected.
Conclusions:
- This study represents the first application of SAGE to characterize gene expression in normal human trabecular meshwork.
- SAGE analysis offers an unbiased method for sampling gene expression in biological tissues.
- The generated data offer valuable insights into the molecular biology of human aqueous outflow and glaucoma.
