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A Enriquez de Salamanca, Y Diebold, M Calonge, S Callejo, ME Stern; Immunodetection of Muscarinic and Adrenergic Receptors in an Epithelial Cell Line from Human Conjunctiva . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3166.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To determine the presence of muscarinic and α- and ß- adrenergic receptor subtypes in cultured epithelial cells from human conjunctiva. Methods: A conjunctival epithelial cell line (NHC) obtained in our laboratory was used. Cells in 62-65 passages were seeded onto glass coverslips and fixed with methanol for 10 min at -20ºC. The presence of neurotransmitter receptors was analyzed by indirect immunofluorescence with specific antibodies. Muscarinic (M1, M2, M3) and α- and ß- adrenergic (α1A-, α1B-, α1C-, α2A-, α2B-, α2C-, ß1-, ß2-, ß3-) receptor subtypes were studied. Negative controls included the omission of primary antibodies. Human conjunctiva and rodent ocular tissue sections were used as positive controls. Results: M2- and M3- muscarinic receptors were immunodetected in cytosol of NHC cells. M1- muscarinic receptor was not immunodetected. α1C- and α2A- and α2B-adrenergic receptor subtypes were also immunodetected in the cytosol of NHC cells. No immunoreactivity was found for α1A- , α1B- and α2C- adrenergic receptors. ß1- and ß2- adrenergic receptor subtypes were immunodetected, showing a predominant cytosolic distribution. ß3 adrenergic receptor was not immunodetected. Conclusion: NHC cell line from human conjunctiva express M2- and M3-muscarinic and α1C-, α2A-, α2B-, ß1- and ß2- adrenergic receptor subtypes. Therefore, NHC cells may be a useful in vitro system to study neural regulation of conjunctival cell functioning in some ocular surface diseases.
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