{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,26]],"date-time":"2025-09-26T22:18:02Z","timestamp":1758925082041},"reference-count":49,"publisher":"The Endocrine Society","issue":"10","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2008,10,1]]},"abstract":"<jats:p>In neuroendocrine cells, discharge of hormones follows the fusion of exocytotic vesicles with the plasma membrane at confined sites; however, the molecular nature of these distinct sites remains poorly understood. We studied intact pituitary lactotrophs and plasma membrane lawns by confocal microscopy in conjunction with antibodies against rat prolactin (rPRL), soluble N-ethylmaleimide-sensitive factor-attachment protein receptor (SNARE) proteins (syntaxin-1 and synaptobrevin-2,) and fluorescent cholera toxin subunit B (CT-B), a marker of ganglioside monosialic acid (GM1) lipid rafts, to examine 1) whether rPRL vesicles discharge cargo at GM1 rafts, 2) whether discharging rPRL vesicles interact with SNAREs, and 3) to examine the overlap of GM1 rafts, rPRL, and syntaxin-1 sites in plasma membrane lawns. In intact cells, immunofluorescently labeled rPRL poorly colocalized (&amp;lt;6%) with CT-B. In conditions favoring endocytotic trafficking, vesicle SNARE synaptobrevin-2 modestly colocalized (35%) with CT-B, whereas it highly colocalized (58%) with retrieved rPRL. Although partial mixing between rPRL and CT-B intracellular trafficking pathways is likely, our results indicated that rPRL discharge involves interactions with plasma membrane SNAREs, but not with GM1 rafts. In support of this, the plasma membrane SNARE syntaxin-1 poorly colocalized with CT-B (&amp;lt;5%), whereas it highly colocalized (75%) with rPRL in inside-out plasma membrane lawns. Spontaneous and stimulated rPRL discharge in live lactotrophs is thus associated with plasma membrane sites enriched with SNARE proteins, however, spatially confined to plasma membrane areas other than GM1 rafts.<\/jats:p>","DOI":"10.1210\/en.2008-0096","type":"journal-article","created":{"date-parts":[[2008,6,13]],"date-time":"2008-06-13T01:54:08Z","timestamp":1213322048000},"page":"4948-4957","source":"Crossref","is-referenced-by-count":21,"title":["Prolactin Secretion Sites Contain Syntaxin-1 and Differ from Ganglioside Monosialic Acid Rafts in Rat Lactotrophs"],"prefix":"10.1210","volume":"149","author":[{"given":"Paula P.","family":"Gonc\u0327alves","sequence":"first","affiliation":[{"name":"Centro de Estudos do Ambiente e do Mar (P.P.G.), Departamento de Biologia, Universidade de Aveiro, 3810-193 Aveiro, Portugal"}]},{"given":"Matjaz\u030c","family":"Stenovec","sequence":"first","affiliation":[{"name":"Celica Biomedical Center (M.S., H.H.C., S.G., M.K., R.Z.), Technology Park Ljubljana, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia"},{"name":"Laboratory of Neuroendocrinology-Molecular Cell Physiology (M.S., H.H.C., S.G., M.K., R.Z.), Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia"}]},{"given":"Helena H.","family":"Chowdhury","sequence":"first","affiliation":[{"name":"Celica Biomedical Center (M.S., H.H.C., S.G., M.K., R.Z.), Technology Park Ljubljana, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia"},{"name":"Laboratory of Neuroendocrinology-Molecular Cell Physiology (M.S., H.H.C., S.G., M.K., R.Z.), Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia"}]},{"given":"Sonja","family":"Grilc","sequence":"first","affiliation":[{"name":"Celica Biomedical Center (M.S., H.H.C., S.G., M.K., R.Z.), Technology Park Ljubljana, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia"},{"name":"Laboratory of Neuroendocrinology-Molecular Cell Physiology (M.S., H.H.C., S.G., M.K., R.Z.), Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia"}]},{"given":"Marko","family":"Kreft","sequence":"first","affiliation":[{"name":"Celica Biomedical Center (M.S., H.H.C., S.G., M.K., R.Z.), Technology Park Ljubljana, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia"},{"name":"Laboratory of Neuroendocrinology-Molecular Cell Physiology (M.S., H.H.C., S.G., M.K., R.Z.), Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia"}]},{"given":"Robert","family":"Zorec","sequence":"first","affiliation":[{"name":"Celica Biomedical Center (M.S., H.H.C., S.G., M.K., R.Z.), Technology Park Ljubljana, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia"},{"name":"Laboratory of Neuroendocrinology-Molecular Cell Physiology (M.S., H.H.C., S.G., M.K., R.Z.), Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia"}]}],"member":"80","published-online":{"date-parts":[[2008,6,12]]},"reference":[{"key":"2020071612264323900_R1","doi-asserted-by":"crossref","first-page":"17221","DOI":"10.1074\/jbc.R000005200","article-title":"Structure and function of sphingolipid- and cholesterol-rich membrane rafts.","volume":"275","author":"Brown","year":"2000","journal-title":"J Biol Chem"},{"key":"2020071612264323900_R2","doi-asserted-by":"crossref","first-page":"412","DOI":"10.1016\/S0166-2236(02)02215-4","article-title":"Lipid rafts in neuronal signaling and function.","volume":"25","author":"Tsui-Pierchala","year":"2002","journal-title":"Trends Neurosci"},{"key":"2020071612264323900_R3","doi-asserted-by":"crossref","first-page":"3734","DOI":"10.1073\/pnas.96.7.3734","article-title":"Raft association of SNAP receptors acting in apical trafficking in Madin-Darby canine kidney cells.","volume":"96","author":"Lafont","year":"1999","journal-title":"Proc Natl Acad Sci USA"},{"key":"2020071612264323900_R4","doi-asserted-by":"crossref","first-page":"5619","DOI":"10.1073\/pnas.091502398","article-title":"SNARE proteins are highly enriched in lipid rafts in PC12 cells: implications for the spatial control of exocytosis.","volume":"98","author":"Chamberlain","year":"2001","journal-title":"Proc Natl Acad Sci USA"},{"key":"2020071612264323900_R5","doi-asserted-by":"crossref","first-page":"49750","DOI":"10.1074\/jbc.M206936200","article-title":"The vesicle- and target-SNARE proteins that mediate Glut4 vesicle fusion are localized in detergent-insoluble lipid rafts present on distinct intracellular membranes.","volume":"277","author":"Chamberlain","year":"2002","journal-title":"J Biol Chem"},{"key":"2020071612264323900_R6","doi-asserted-by":"crossref","first-page":"1236","DOI":"10.1074\/jbc.M410674200","article-title":"The SNARE proteins SNAP-25 and SNAP-23 display different affinities for lipid rafts in PC12 cells. 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