January 1999
Volume 40, Issue 1
Free
Articles  |   January 1999
Expression of alternatively spliced growth factor receptor isoforms in the human trabecular meshwork.
Author Affiliations
  • R J Wordinger
    Department of Anatomy and Cell Biology and The North Texas Eye Research Institute, University of North Texas Health Science Center at Fort Worth, 76107-2699, USA.
  • A F Clark
    Department of Anatomy and Cell Biology and The North Texas Eye Research Institute, University of North Texas Health Science Center at Fort Worth, 76107-2699, USA.
  • R Agarwal
    Department of Anatomy and Cell Biology and The North Texas Eye Research Institute, University of North Texas Health Science Center at Fort Worth, 76107-2699, USA.
  • W Lambert
    Department of Anatomy and Cell Biology and The North Texas Eye Research Institute, University of North Texas Health Science Center at Fort Worth, 76107-2699, USA.
  • S E Wilson
    Department of Anatomy and Cell Biology and The North Texas Eye Research Institute, University of North Texas Health Science Center at Fort Worth, 76107-2699, USA.
Investigative Ophthalmology & Visual Science January 1999, Vol.40, 242-247. doi:
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    • Get Citation

      R J Wordinger, A F Clark, R Agarwal, W Lambert, S E Wilson; Expression of alternatively spliced growth factor receptor isoforms in the human trabecular meshwork.. Invest. Ophthalmol. Vis. Sci. 1999;40(1):242-247.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

PURPOSE: Growth factors act through high-affinity cell surface receptors expressed by target cells and are critical modulators of cell function. Because aqueous humor is known to contain growth factors, these molecules may play a key role in maintaining the normal function of the human trabecular meshwork (HTM). Alternate mRNA splicing is an important mechanism used by cells to generate diverse isoforms of growth factor receptors. Although previous investigators have suggested that HTM cells may express alternative isoforms of several growth factor receptors, there have been no studies to verify these preliminary findings. The objective of this study was to determine whether cultured and ex vivo HTM cells express alternate isoforms of hepatocyte, keratinocyte, and transforming growth factor beta (TGFbeta)-II receptors and to characterize the isoform molecular sequences. METHODS: To determine whether cells within the HTM express mRNA for alternate isoforms of growth factor receptors, total RNA was isolated from several well-characterized HTM cell lines that were established from donors of various ages and from fresh ex vivo HTM tissues from healthy donors. After cDNA synthesis, polymerase chain reaction was initiated using specific primers for alternate forms of the following receptors: hepatocyte growth factor (HGFR), keratinocyte growth factor (KGFR), and transforming growth factor beta receptor II (TGFbetaR-II). Specificity and characterization of the polymerase chain reaction amplification products were determined by nucleic acid sequencing. RESULTS: Amplification products of the expected size for the growth factor isoforms were expressed in cell lines and in ex vivo tissues. Nucleic acid sequencing showed that cultured HTM cells and fresh ex vivo trabecular meshwork tissues expressed specific mRNA for alternatively spliced isoforms of HGFR, KGFR, and TGFbetaR-II The HGFR alternate isoform contained a 96-bp insert in the C-terminal coding region of the cytoplasmic tyrosine kinase domain. The KGFR alternate isoform is a soluble, truncated form, because it has no transmembrane or cytoplasmic domain as does the normal membrane-associated form. The TGFbetaR-II alternate isoform contained a 75-bp insert in the N-terminal coding region of the extracellular domain. CONCLUSIONS: In vitro and ex vivo HTM cells express mRNA for alternatively spliced isoforms of HGFR, KGFR, and TGFbetaR-II. These alternatively spliced receptor isoforms may be functional within the HTM and may play a critical role in maintaining the normal microenvironment of this important tissue.

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