title: Integrin adhesion in regulation of lacrimal gland acinar cell secretion.
abstract: The extracellular microenvironment regulates lacrimal gland acinar cell secretion. Culturing isolated rabbit lacrimal gland acinar cells on different extracellular matrix proteins revealed that laminin enhances carbachol-stimulated secretion to a greater extent than other extracellular matrix proteins investigated. Furthermore, immunofluorescence indicated that integrin subunits, potentially functioning as laminin receptors are present in acinar cells. Among these, the integrin alpha6 and beta1 subunit mRNA expression was also confirmed by RT-PCR and sequence analysis. Secretion assays, which measured beta-hexosaminidase activity released in the culture media, demonstrated that function-blocking integrin alpha6 and beta1 monoclonal antibodies (mAbs) induce a rapid, transient and dose-dependent secretory response in cultured cells. To determine the intracellular pathways by which integrin alpha6 and beta1 mAbs could induce secretion, selected second messenger molecules were inhibited. Although inhibitors of protein kinase C and IP(3)-induced Ca(2+) mobilization attenuated carbachol-stimulated secretion, no effect on integrin mAb-induced release was observed. In addition, protein tyrosine kinases do not appear to have a role in transducing signals arising from mAb interactions. Our data clearly demonstrate, though, that cell adhesion through integrins regulates secretion from lacrimal gland acinar cells. The fact that the integrin mAbs affect the cholinergic response differently and that the integrin beta1 mAb secretion, but not the alpha6, was attenuated by the phosphatase inhibitor, sodium orthovanadate, suggests that each subunit utilizes separate intracellular signaling pathways to induce exocytosis. The results also indicate that the secretory response triggered by the beta1 integrin mAb is generated through dephosphorylation events.
authors.affiliation: Department of Chemistry and Biomedical Sciences, University of Kalmar, SE-391 82 Kalmar, Sweden.
authors.forename: Sofia V
authors.initials: SV
authors.lastname: Andersson
authors.forename: Sarah F
authors.initials: SF
authors.lastname: Hamm-Alvarez
authors.forename: J Peter
authors.initials: JP
authors.lastname: Gierow
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002217:Carbachol / Q000494:pharmacology; D002448:Cell Adhesion; D002478:Cells, Cultured; D018679:Cholinergic Agonists / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism*; D039503:Integrin alpha6 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D019012:Integrin beta1 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D016023:Integrins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D007765:Lacrimal Apparatus / Q000166:cytology / Q000378:metabolism*; D007797:Laminin / Q000378:metabolism* / Q000494:pharmacology; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014638:Vanadates / Q000494:pharmacology
chemical_list: D000911:Antibodies, Monoclonal; D018679:Cholinergic Agonists; D039503:Integrin alpha6; D019012:Integrin beta1; D016023:Integrins; D007797:Laminin; D012333:RNA, Messenger; D014638:Vanadates; D002217:Carbachol; D011494:Protein Kinases; D011493:Protein Kinase C; D010744:Phosphoric Monoester Hydrolases; D002118:Calcium