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Preproinsulin mRNA in the rat eye

G C Budd1, B Pansky, L Glatzer

  • 1Department of Physiology, Medical College of Ohio, Toledo 43699.

Investigative Ophthalmology & Visual Science
|February 1, 1993
PubMed
Summary

Researchers confirmed insulin gene expression in rat eyes, specifically in the retina. This finding supports previous evidence of insulin-related peptides in the rat retina and cultured retinal cells.

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Area of Science:

  • Ophthalmology
  • Molecular Biology
  • Genetics

Background:

  • Previous studies indicated the presence of insulin-related peptides in the rat retina using immunoassay, immunocytochemistry, and in situ hybridization.
  • The expression of insulin genes in the rat eye remained to be confirmed at the molecular level.

Purpose of the Study:

  • To confirm the expression of insulin genes in the rat eye.
  • To extend previous findings on insulin-related peptides in the rat retina.

Main Methods:

  • RNA isolation from whole rat eyes and specific ocular tissues (retina, choroid, iris, lens, vitreous).
  • Preparation of insulin-specific complementary DNA (cDNA) using a coupled reverse transcription polymerase chain reaction (RT-PCR) protocol.
  • Characterization and identification of amplified cDNA products via Southern transfers, restriction fragment analyses, and nucleotide sequencing.

Main Results:

  • Amplified cDNA fragments of 329 ± 6 base pairs (bp) were obtained from whole rat eye and retina RNA.
  • Southern blot analysis showed homology between retinal cDNA and rat pancreatic preproinsulin1 DNA.
  • Nucleotide sequencing confirmed the amplified cDNA was identical to rat pancreatic preproinsulin1 DNA.

Conclusions:

  • The presence of preproinsulin1 messenger RNA (mRNA) in the rat retina was confirmed.
  • This molecular confirmation aligns with prior studies demonstrating insulin immunoreactivity and in situ hybridization evidence of insulin-related mRNA in retinal glial cells.

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