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S. K. Carlsson, A. Pettersson, J. P. Gierow, D. Delbro; Presence of SLURP-1 and Nicotinic Acetylcholine Receptor in the Rabbit Lacrimal Gland. Invest. Ophthalmol. Vis. Sci. 2009;50(13):4242.
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It has recently been shown that nicotinic acetylcholine receptors are present also in non-neuronal tissue and that they together with SLURP (secreted mammalian Ly-6/urokinase plasminogen acivator receptor-related protein) might be involved in immuno-modulatory/apoptotic events. The aim of the present study is therefore to investigate the presence of the alpha-7 subunit of the nicotinic acetylcholine receptor and SLURP in the rabbit lacrimal gland.
Rabbit lacrimal gland acinar cells were prepared according to our standard procedure (Gierow et al., Am. J. Physiol. (Cell Physiology) 271: C1685, 1996; Andersson et al, Glycobiol. 15: 211, 2005) yielding single cells that were placed in a serum-free culture medium on standard culture plates coated with Matrigel for two days. The cultured cells were rinsed briefly and secretion was measured after a pre-incubation for 30 min at 37 C, with buffer alone, followed by incubation for an additional 30 min w./w.o. secretagogues as indicated. Basal secretion and regulated secretion was determined enzymatically as secreted beta-hexosaminidase activity. The lacrimal gland was also subjected to immunohistochemistry to confirm the presence the alpha-7 subunit, SLURP-1 and -2.
Secretion of beta-hexosaminidase from cultured acinar cells was significantly increased by approximately 30 %, at 1 µM nicotine. Immunohistochemistry indicated that both the alpha-7 subunit of the nicotinic acetylcholine receptor and its ligand SLURP-1 are present in lacrimal tissue, whereas SLURP-2 could not be detected.
Our results indicate that both the alpha-7 subunit of the nicotinic acetylcholine receptor and its ligand SLURP-1 are present in the rabbit lacrimal gland and that nicotine tartrate has stimulatory effects on protein secretion in cultured cells.
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