September 1993
Volume 34, Issue 10
Free
Articles  |   September 1993
Nuclear muscarinic acetylcholine receptors in corneal cells from rabbit.
Author Affiliations
  • G J Lind
    Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas 75235-9057.
  • H D Cavanagh
    Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas 75235-9057.
Investigative Ophthalmology & Visual Science September 1993, Vol.34, 2943-2952. doi:
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      G J Lind, H D Cavanagh; Nuclear muscarinic acetylcholine receptors in corneal cells from rabbit.. Invest. Ophthalmol. Vis. Sci. 1993;34(10):2943-2952.

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Abstract

PURPOSE: Previous studies have indicated that muscarinic acetylcholine receptors (mAChR) may be present in an unexpected, unique location and play a singular role in cellular growth regulation of rabbit corneal epithelium that may be of general physiologic significance if found in other cells. The purpose of this study was to examine rabbit corneas and corneal cells in culture to determine mAChR location and tissue distribution. METHODS: Using [3H]-propylbenzilylcholine mustard ([3H]PrBChM), which binds covalently to the active site of mAChR, rabbit corneal cross-sections, cultured corneal keratocytes, epithelial and endothelial cells, as well as nuclei isolated from these cultured corneal cells were labeled, stained, and autoradiographed. Nuclei labeled with [3H]PrBChM were further analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. RESULTS: Direct visual confirmation of the localization of mAChRs was obtained. MAChR were found in epithelial and endothelial layers of fresh-frozen corneal cross-sections, in cultured rabbit epithelial and endothelial cells, and on isolated rabbit epithelial and endothelial cell nuclei. mAChR were not detectable in keratocytes with these techniques. When [3H]PrBChM-labeled nuclei from cultured corneal cells were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, epithelial and endothelial samples showed specific mAChR binding, whereas binding to keratocyte nuclei was not detectable. CONCLUSIONS: As a result of these findings, a revised hypothesis is suggested for the locations and possible functions of mAChR in regulation of growth in corneal and other cells.

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