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John L. Ubels, Ilene K. Gipson, Sandra J. Spurr-Michaud, Ann S. Tisdale, Rachel E. Van Dyken, Mark H. Hatton; Analysis of Gene Expression in Accessory Lacrimal Glands by Laser Microdissection and cDNA Microarrays. Invest. Ophthalmol. Vis. Sci. 2012;53(14):2702.
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© ARVO (1962-2015); The Authors (2016-present)
The accessory lacrimal glands are assumed to contribute to the production of tear fluid, but little is known about their function. The goal of this study was to conduct an analysis of gene expression by glands of Wolfring that would provide a more complete picture of the function of these glands.
Accessory gland cells were isolated from frozen sections of human eyelids by laser microdissection. Acinar cells were also captured from human main lacrimal glands. RNA was extracted from the cells and amplified using the NuGEN Ovation V2 Amplification System. cDNA was hybridized to Affymetrix Human Genome U133 Plus 2.0 3’ gene expression arrays. Gene lists generated from expression scans using dChip software were analyzed using DAVID Functional AnnotationBioinformatics Microarray Analysis software.
Cluster analysis of 4 accessory glands and 2 main glands grouped the glands of Wolfring separately from main lacrimal glands, with 164 genes expressed at least 3X higher in accessory glands. The gene expressed most highly in accessory glands compared to the main gland is a mucin-like glycoprotein previously identified only in mammary epithelium and salivary gland. Other genes of relevance to lacrimal function that were strongly expressed in accessory glands at levels above the main gland include beta-defensin, proenkephlin,carbonic anhydrase IV and muscarinic cholinergic receptor 1 (mAChR). Accessory glands also expressed many other genes of known relevance to lacrimal function, including lacritin, lipocalin, lactoferrin, lysozyme, prolactin receptor, alpha-adrenergic receptor, aquaporins (AQP) and many ion channels and transporters. Expression of AQP5, mAChR, lacritin, lipocalin, lysozyme and lactoferrrin was confirmed by immunofluorescence.
This is the first large scale genetic analysis of glands of Wolfring. Several notable differences between accessory and main lacrimal glands were detected, although the data must be interpreted with caution due to the limited number of glands available. The data suggest that in general the function of glands of Wolfring is similar to main lacrimal glands. Since these glands secrete directly onto the ocular surface, their location may allow rapid response to exogenous stimuli and makes them readily accessible to topical drugs.
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