Expression of Neural Receptors in Mouse Meibomian Gland

Purpose: To investigate gene and protein expression profiles of neural receptors found in the mouse meibomian gland. RNA and protein levels were determined for neuropeptide Y (NPY) receptor, vasoactive intestinal peptide (VIP) receptor, substance P (SP) receptor, and muscarinic cholinergic receptor (mAChR) subtypes M1-M5 in the mouse meibomian gland. Methods: Frozen sections of Balb/c mouse eyelids were subjected to laser capture microdissection to isolate pure samples of meibomian gland ductal and acinar cells. Real-time polymerase chain reaction, immunolabeling, and Western blot analysis for SP receptor, VIP receptor, NPY receptor, and mAChR subtypes M1-M5 were performed on meibomian gland ductal and acinar cells. Results: Expression of NPY1 receptor, VIP receptor 1, SP receptor, and all 5 mAChR subtypes was found in all meibomian gland ductal and acinar cells analyzed by real-time polymerase chain reaction. Immunolabeling and Western blot analysis confirmed the presence of NPY1 receptor, VIP receptor 1, SP receptor, and all 5 mAChR subtypes in the meibomian gland. The levels were variable with the duct showing greater levels of NPY1 receptor, SP receptor, and mAChRs 1, 2, 4, and 5 than with the gland. Conclusions: VIP receptor 1, SP receptor, NPY1 receptor, and mAChR subtypes may be involved in the regulation of meibomian gland secretion. Laser capture microdissection in conjunction with gene expression analysis provides an excellent approach for studying meibomian gland cells about which relatively little is known at the molecular level.