title: Expression of two membrane fusion proteins, synaptosome-associated protein of 25 kDa and vesicle-associated membrane protein, in choroid plexus epithelium.
abstract: In addition to being the major site of cerebrospinal fluid formation, the choroid plexus epithelium emerges as an important source of polypeptides in the brain. Physiologically regulated release of some polypeptides synthesized by the choroid plexus has been shown. The molecular mechanisms underlying this polypeptide secretion have not been characterized, however. In the present study, synaptosome-associated protein of 25 kDa and vesicle-associated membrane protein, two membrane fusion proteins playing a critical role in exocytosis in neurons and endocrine cells, were found to be expressed in the choroid plexus epithelium. It was also shown that in choroidal epithelium, synaptosome-associated protein of 25 kDa and vesicle-associated membrane protein stably interact. Two members of the vesicle-associated membrane protein family, vesicle-associated membrane protein-1 and vesicle-associated membrane protein-2, were expressed in the rat choroid plexus at the messenger RNA and protein level. However, their newly discovered isoforms, vesicle-associated membrane protein-1b and vesicle-associated membrane protein-2b, produced by alternative RNA splicing, were not detected in choroidal tissue. Immunohistochemistry demonstrated that vesicle-associated membrane protein is confined to the cytoplasm of choroidal epithelium, whereas synaptosome-associated protein of 25 kDa is associated with plasma membranes, albeit with a varied cellular distribution among species studied. Specifically, in the rat choroid plexus, synaptosome-associated protein of 25 kDa was localized to the basolateral membrane domain of choroidal epithelium and was expressed in small groups of cells. In comparison, in ovine and human choroidal tissues, apical staining for synaptosome-associated protein of 25 kDa was found in the majority of epithelial cells. These species-related differences in cellular synaptosome-associated protein of 25 kDa distribution suggested that the synaptosome-associated protein of 25 kDa homologue, synaptosome-associated protein of 23 kDa, is also expressed in the rat choroid plexus, which was confirmed by reverse-transcriptase polymerase chain reaction. Our findings suggest that synaptosome-associated protein of 25 kDa and vesicle-associated membrane protein are involved in secretion of polypeptides from the choroid plexus epithelium. The presence of synaptosome-associated protein of 25 kDa and its homologue as well as multiple isoforms of vesicle-associated membrane protein in choroidal epithelium may play a role in the apical versus basolateral targeting of secretory vesicles.
authors.affiliation: Department of Clinical Neurosciences, Brown University School of Medicine, Aldrich Building, Room 403, 593 Eddy Street, Providence, RI 02903, USA.
authors.forename: I
authors.initials: I
authors.lastname: Chung
authors.forename: A
authors.initials: A
authors.lastname: Burkart
authors.forename: J
authors.initials: J
authors.lastname: Szmydynger-Chodobska
authors.forename: K A
authors.initials: KA
authors.lastname: Dodd
authors.forename: W S
authors.initials: WS
authors.lastname: Trimble
authors.forename: K V
authors.initials: KV
authors.lastname: Miller
authors.forename: M
authors.initials: M
authors.lastname: Shim
authors.forename: A
authors.initials: A
authors.lastname: Chodobski
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002831:Choroid Plexus / Q000737:chemistry / Q000502:physiology*; D004848:Epithelium / Q000737:chemistry / Q000502:physiology; D005089:Exocytosis / Q000502:physiology; D007150:Immunohistochemistry; D008297:Male; D008561:Membrane Fusion / Q000502:physiology; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics*; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000235:genetics*; D011233:Precipitin Tests; D050683:R-SNARE Proteins; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050825:Synaptosomal-Associated Protein 25
chemical_list: D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D050683:R-SNARE Proteins; D012333:RNA, Messenger; C494210:Snap25 protein, rat; D050825:Synaptosomal-Associated Protein 25