Cytoplasmic surface ultrastructures of gap junctions in bovine lens fibers

T. Hatae, H. Iida, A. Kuraoka, Y. Shibata

Research output: Contribution to journalArticle

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Abstract

Purpose. To examine the cytoplasmic surface ultrastructures of lens fiber gap junctions, where the cytoplasmic domains of connexons were expected to be exposed. Methods. Bovine lens fiber gap junctions, both in situ and in the form of isolated membranes, were examined with the deep etching replica methods. Isolated membranes were also examined with the same methods after the treatment with endoprotemase glu-C4 which is known to cleave off the cytoplasmic domain of a putative lens fiber connexin MP70 to determine whether any structural changes should occur between proteolyzed and nonproteolyzed gap junctions. In addition, both proteolyzed and nonproteolyzed gap junctions were studied by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunolabeling with the monoclonal antibody that recognized cytoplasmic domain of MP70 to clarify whether MP70 lost its cytoplasmic domain by the treatment with endoproteinase glu-C. Results. Gap junctions were shown to have particulate substructures on their cytoplasmic surfaces; the distributions of the particles were restricted within gap junctional plaques and the non-gap-junctional areas showed smooth cytoplasmic surfaces. Although the treatment with endoproteinase glu-C failed to remove the cytoplasmic particles of gap junctions in deep etching replica study, MP70 was shown to have lost its cytoplasmic domain in sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunolabeling studies. Conclusions. Each particle revealed on the cytoplasmic surfaces of lens fiber gap junctions corresponded to the cytoplasmic domain of a connexon. The particles were not removed by the treatment with endoproteinase glu-C, whereas MP70 was cleaved by the same treatment.

Original languageEnglish
Pages (from-to)2164-2173
Number of pages10
JournalInvestigative Ophthalmology and Visual Science
Volume34
Issue number7
Publication statusPublished - Jan 1 1993

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Gap Junctions
Lenses
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Connexins
Membranes
Monoclonal Antibodies
glutamyl endopeptidase

All Science Journal Classification (ASJC) codes

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Cytoplasmic surface ultrastructures of gap junctions in bovine lens fibers. / Hatae, T.; Iida, H.; Kuraoka, A.; Shibata, Y.

In: Investigative Ophthalmology and Visual Science, Vol. 34, No. 7, 01.01.1993, p. 2164-2173.

Research output: Contribution to journalArticle

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