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Development of Conjunctival Goblet Cells and Their Neuroreceptor Subtype Expression

From the Schepens Eye Research Institute and Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts.

PURPOSE. To investigate expression of muscarinic, cholinergic, and adrenergic receptors on developing conjunctival goblet cells.

METHODS. Eyes were removed from rats 9 to 60 days old, fixed, and used for microscopy. For glycoconjugate expression, sections were stained with Alcian blue/periodic acid-Schiff’s reagent (AB/PAS) and with the lectins Ulex europeus agglutinin I (UEA-I) and Helix pomatia agglutinin (HPA). Goblet cell bodies were identified using anti-cytokeratin 7 (CK7). Nerve fibers were localized using anti-protein gene product 9.5. Location of muscarinic and adrenergic receptors was investigated using anti-muscarinic and ß-adrenergic receptors.

RESULTS. At days 9 and 13, single apical cells in conjunctival epithelium stained with AB/PAS, UEA-I, and CK7. At days 17 and 60, increasing numbers of goblet cells were identified by AB/PAS, UEA-I, HPA, and CK7. Nerve fibers were localized around stratified squamous cells and at the epithelial base at days 9 and 13, and around goblet cells and at the epithelial base at days 17 and 60. At days 9 and 13, M₂- and M₃-muscarinic and ß₂-adrenergic receptors were found in stratified squamous cells, but M₁-muscarinic and ß₁-adrenergic receptors were not detected. At days 17 and 60, M₂- and M₃-muscarinic receptors were found in goblet cells, whereas M₁-muscarinic receptors were in stratified squamous cells. ß₁- and ß₂-Adrenergic receptors were found on both cell types. ß₃-Adrenergic receptors were not detected.

CONCLUSIONS. In conjunctiva, nerves, M₂- and M₃-muscarinic, and ß₁- and ß₂-adrenergic receptors are present on developing goblet cells and could regulate secretion as eyelids open.

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