title: Increase in neuropilin-1 on the surface of growth cones and putative raft domains in neuronal NG108-15 cells co-cultured with vascular smooth muscle SM-3 cells.
abstract: The mechanisms underlying autonomic innervation to its targets involve various chemical factors, but have not yet been elucidated in detail. We constructed a co-culture system of neuronal cells and vascular smooth muscle cells to investigate the mechanisms underlying innervation of the vasculature. A co-culture with the vascular smooth muscle cell line, SM-3 significantly promoted cell viability, neurite extension, and neuropilin-1 (Nrp-1) mRNA expression in the cholinergic neuronal cell line, NG108-15. Furthermore, immunocytochemistry with or without a detergent treatment revealed that a co-culture with SM-3 cells or culturing with the conditioned medium of SM-3 cells translocated Nrp-1 onto the cell surface of growth cones rather than varicosities of NG108-15 cells. Immunofluorescent microscopy combined with a cold detergent treatment or cholesterol depletion revealed that Nrp-1 accumulated in putative raft domains in the plasma membrane of NG108-15 cells co-cultured with SM-3 cells. The results of the present study suggest that some soluble factors from smooth muscle cells may affect the localization of Nrp-1 in cholinergic neuronal cells, which may, in turn, be involved in the autonomic innervation of blood vessels.
authors.affiliation: Department of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-Ku, Kyoto, 606-8585, Japan, ryoshimu@kit.ac.jp.
authors.forename: Ryoichi
authors.initials: R
authors.lastname: Yoshimura
authors.forename: Ayumi
authors.initials: A
authors.lastname: Kyuka
authors.forename: Miwa
authors.initials: M
authors.lastname: Jinno
authors.forename: Satomi
authors.initials: S
authors.lastname: Nishio
authors.forename: Mamoru
authors.initials: M
authors.lastname: Matsusaka
authors.forename: Tomoki
authors.initials: T
authors.lastname: Nishida
authors.forename: Yasuhisa
authors.initials: Y
authors.lastname: Endo
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D018920:Coculture Techniques; D015870:Gene Expression; D020439:Growth Cones / Q000378:metabolism*; D007150:Immunohistochemistry; D021962:Membrane Microdomains / Q000378:metabolism*; D051379:Mice; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D039942:Neuropilin-1 / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats
chemical_list: D012333:RNA, Messenger; D039942:Neuropilin-1