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Related Experiment Videos

Quantification of MUC2 and MUC5AC transcripts in human conjunctiva.

R W McKenzie1, J E Jumblatt, M M Jumblatt

  • 1Department of Ophthalmology and Visual Sciences, University of Louisville, School of Medicine, Kentucky 40292, USA.

Investigative Ophthalmology & Visual Science
|March 11, 2000
PubMed
Summary

MUC2 transcripts are present in human conjunctival tissue, though at much lower levels than MUC5AC. This study utilized quantitative PCR to detect these ocular mucin transcripts.

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Area of Science:

  • Ophthalmology
  • Molecular Biology
  • Biochemistry

Background:

  • Mucins are key components of the ocular surface.
  • MUC5AC is a known conjunctival goblet cell mucin.
  • MUC2, a goblet cell mucin from the intestine, has not been previously detected in conjunctival tissue.

Purpose of the Study:

  • To determine the presence and quantify the levels of MUC2 and MUC5AC transcripts in normal human conjunctiva.
  • To compare the relative expression levels of these mucins.

Main Methods:

  • Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was employed.
  • Internal standards were used for accurate quantification of MUC2 and MUC5AC mRNA.
  • Expression levels were normalized to beta2-microglobulin (beta2M).

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Main Results:

  • MUC5AC transcripts were detected at levels 10-fold lower than beta2M.
  • MUC2 transcripts were found to be present, contrary to previous reports.
  • MUC2 expression was significantly lower than MUC5AC, approximately 5900-fold less.

Conclusions:

  • MUC2 transcripts are present in human conjunctival tissue, albeit at very low levels compared to MUC5AC.
  • This study is the first to quantitatively analyze mucin expression in human ocular tissue using competitive PCR.
  • The sensitive method enabled detection of MUC2 transcripts missed by other techniques, allowing for relative expression comparisons.