Polarized targeting of IgLON cell adhesion molecule OBCAM to dendrites in cultured neurons.
Opioid-binding cell adhesion molecule (OBCAM) belongs to the immunoglobulin superfamily CAMs and shows a dendritically polarized distribution in hypothalamic magnocellular neurons. In the present study, the cellular localization of OBCAM was monitored in cultured cortical and hippocampal neurons to examine its polarized distribution. Double labeling immunofluorescence microscopy after fixation showed only faint OBCAM immunoreactivity in the neuronal somata during the early stages of culture, whereas the immunoreactivity was strong in MAP2-positive somata and dendrites of fully polarized neurons after longer culture. Moreover, the immunoreactivity for OBCAM showed a punctate pattern in the dendrites similar to the immunostaining pattern of synapsin I. High resolution revealed close apposition with only a partial overlap of synapsin I and OBCAM immunoreactivities, suggesting the synaptic localization of OBCAM to the dendrites. When the fully polarized neurons were reacted with anti-OBCAM antibody before fixation, OBCAM immunoreactivity became stronger on the dendritic surface than the somatic surface. Extracellular immunoreactivity was eliminated with phosphatidylinositol-specific phospholipase C and this immunoreactivity resisted extraction with the nonionic detergent Triton X-100 at 4 degrees C, indicating that OBCAM is attached to the rafts via a glycosylphosphatidyl inositol anchor. These results indicate that OBCAM is efficiently targeted to the dendritic surface of fully polarized cortical and hippocampal neurons. OBCAM is, hence, concluded to be a dendrite-associated CAM in cortical and hippocampal neurons as in hypothalamic magnocellular neurons.
Miyata
Seiji
S
Department of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, 606-8585, Kyoto, Japan. smiyata@ipc.kit.ac.jp
Matsumoto
Naoko
N
Maekawa
Shohei
S
D000818:Animals; D002352:Carrier Proteins / Q000096:biosynthesis*; D015815:Cell Adhesion Molecules / Q000096:biosynthesis*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D003712:Dendrites / Q000648:ultrastructure*; D005455:Fluorescent Antibody Technique; D058851:GPI-Linked Proteins; D006624:Hippocampus / Q000166:cytology; D018613:Microscopy, Confocal; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D017830:Octoxynol; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D054801:Phosphoinositide Phospholipase C; D051381:Rats; D017208:Rats, Wistar; D016704:Synapsins / Q000378:metabolism; D013997:Time Factors; D010738:Type C Phospholipases; D016875:tau Proteins / Q000378:metabolism
D002352:Carrier Proteins; D015815:Cell Adhesion Molecules; D058851:GPI-Linked Proteins; D008869:Microtubule-Associated Proteins; C489215:Opcml protein, rat; D016704:Synapsins; D016875:tau Proteins; D017830:Octoxynol; D010738:Type C Phospholipases; D054801:Phosphoinositide Phospholipase C; D043265:Phosphatidylinositol Diacylglycerol-Lyase