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J.D. Rios–Garcia, D. Zoukhri, D.A. Dartt; Influence Of Age On Mouse Conjunctival Goblet Cell Struture And Function . Invest. Ophthalmol. Vis. Sci. 2004;45(13):1507.
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Purpose:To investigate age–related changes in the structure and function of mouse conjunctival goblet cells. Methods:Fixed sections of conjunctiva, removed from BALB/c mice at 3, 12, 24 months of age, were processed with alcian blue at pH 2.5 and periodic acid Schiff’s reagent (AB–PAS) for morphological and quantitative analyses of conjunctival goblet cells. Immunofluorescence and western blot analyses were conducted to elucidate changes in the amount and composition of goblet cell secretory product. For secretion studies, pieces of inferior conjunctiva were incubated for 30 min in the absence or presence of carbachol (a cholinergic agonist, 10–4 M). Following incubation, the media were collected and the total amount of glycoconjugate released was determined using an enzyme–linked lectin assay (ELLA).Results:As indicated by AB–PAS staning, a reduction in the number of goblet cells in only the superior conjunctiva was observed in 12–24 month–old animals compared to 3 month–old animals. Immunofluorescence analyses using an antibody against MUC5AC and the lectin Helix pomatia agglutinin (HPA), which stain goblet cell secretory products, showed an abnormal pattern of mucin, characterized by dense strands of mucus adhering to the ocular surface of 24 month–old animals. However, in 3 and 12 month–old animals, MUC5AC and HPA lectin were preferentially located in apical secretory granules of conjunctival goblet cells. Western blot analysis from both inferior and superior conjunctiva showed that the MUC5AC antibody and HPA lectin detected a high–molecular weight protein (over to 220 kDa) in all samples. Carbachol induced glycoconjugate secretion in animals of all ages. The secretory response to carbachol did not change with age. Conclusions:We conclude that the number of goblet cells does not vary consistely with age and with mucin secretion. There is potentially a change in the mucus after it is secreted.
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