title: Altered traffic to the lysosome in an ex vivo lacrimal acinar cell model for chronic muscarinic receptor stimulation.
abstract: Evidence suggests that lacrimal and salivary epithelial cells constitutively expose potentially pathogenic autoantigens, but that active regulatory networks normally suppress pathological autoimmune responses . Events that potentially disrupt the regulatory networks include increased exposure of constitutive autoantigens and induced exposure of previously cryptic autoantigen epitopes. Chronic muscarinic receptor (MAChR) stimulation in an ex vivo rabbit lacrimal acinar cell model induces functional and biochemical alterations reminiscent of the functional quiescence associated with Sjogren's syndrome . Chronic MAChR stimulation also elicits changes in the compartmental distribution of beta-hexosaminidase, a product that normally is dually targeted into the lysosomal pathway and the regulated apical secretory pathway. Here, we use subcellular fractionation analyses to further explore the nature of the stimulation-induced traffic changes and to identify effectors that might mediate this change. Overnight stimulation of primary cultured rabbit lacrimal gland acinar cells with 10 microM carbachol (CCh) significantly decreased the abundance of mature cathepsin B in the pre-lysosome and lysosome; decreased the abundance of preprocathepsin B in fractions containing the TGN and late endosome; increased the abundance of procathepsin B in fractions containing the basal-lateral membrane; and increased the accumulation of endocytosed [(125)I]-EGF in the recycling endosome. Alterations in distribution or abundance of traffic effectors included: increased abundances of rab5A and rab6 in the TGN; decreased overall abundance of gamma-adaptin; remarkably increased relative abundance of membrane phase-associated actin; redistribution of cytoplasmic dynein from biosynthetic and proximal endocytic compartments to the lysosome; and redistribution of p150(Glued) from the lysosome to biosynthetic or proximal endocytic compartments. We conclude that chronic MAChR stimulation blocks traffic from the early endosome and the TGN to the lysosome, causing lysosomal proteins to reflux to the TGN, endosomes, and basal-lateral membrane. These traffic alterations may be mediated through action on one or more of the effectors noted.
authors.affiliation: Department of Physiology and Biophysics, Keck School of Medicine, University of Southern California, 1333 San Pablo Street, MMR 626, Los Angeles, CA 90033, USA.
authors.forename: Limin
authors.initials: L
authors.lastname: Qian
authors.forename: Jiansong
authors.initials: J
authors.lastname: Xie
authors.forename: Chadron M
authors.initials: CM
authors.lastname: Rose
authors.forename: Eunbyul
authors.initials: E
authors.lastname: Sou
authors.forename: Hongtao
authors.initials: H
authors.lastname: Zeng
authors.forename: Sarah F
authors.initials: SF
authors.lastname: Hamm-Alvarez
authors.forename: Austin K
authors.initials: AK
authors.lastname: Mircheff
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D001324:Autoantigens / Q000378:metabolism*; D001692:Biological Transport; D015415:Biomarkers / Q000032:analysis; D002217:Carbachol / Q000494:pharmacology; D002401:Cathepsin B / Q000378:metabolism; D002458:Cell Fractionation; D002462:Cell Membrane / Q000378:metabolism*; D018679:Cholinergic Agonists / Q000494:pharmacology; D004398:Dyneins / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism*; D005260:Female; D007765:Lacrimal Apparatus / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism*; D023421:Models, Animal; D011817:Rabbits; D011976:Receptors, Muscarinic / Q000187:drug effects; D013268:Stimulation, Chemical; D000520:alpha-Glucosidases / Q000378:metabolism
chemical_list: D000199:Actins; D001324:Autoantigens; D015415:Biomarkers; D018679:Cholinergic Agonists; D011976:Receptors, Muscarinic; D004815:Epidermal Growth Factor; D002217:Carbachol; D000520:alpha-Glucosidases; D002401:Cathepsin B; D004398:Dyneins