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Preparation of Synaptic Plasma Membrane and Postsynaptic Density Proteins Using a Discontinuous Sucrose Gradient
Published on: September 3, 2014
Synaptic adhesion molecule OBCAM; synaptogenesis and dynamic internalization.
Mayumi Yamada1, Takashi Hashimoto, Noriko Hayashi
1Department of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan.
Brain Research
|July 31, 2007
Summary
Opioid-binding cell adhesion molecule (OBCAM) is crucial for forming new synapses in hippocampal neurons. Its surface levels dynamically change with neuronal activity, impacting synaptic plasticity.
Area of Science:
- Neuroscience
- Cell Biology
- Molecular Biology
Background:
- Opioid-binding cell adhesion molecule (OBCAM) belongs to the IgLON family within the immunoglobulin superfamily.
- Investigating OBCAM's role in neuronal function is essential for understanding synaptic development.
Purpose of the Study:
- To elucidate the functions and dynamics of OBCAM in hippocampal neurons.
- To determine OBCAM's involvement in synaptogenesis and its regulation by neuronal activity.
Main Methods:
- Western blotting to assess OBCAM expression levels over time.
- Double labeling immunofluorescence microscopy to localize OBCAM.
- Functional inhibition using OBCAM-specific antibodies and antisense oligonucleotides.
- Overexpression of OBCAM mRNA via plasmid vectors.
- Investigating OBCAM internalization pathways using 4-aminopyridine and filipin.
Main Results:
- OBCAM expression increased with neuronal culture development.
- OBCAM localized to postsynaptic spines, co-localizing with PSD-95.
- Inhibiting OBCAM function or suppressing its expression reduced synapse formation.
- Overexpressing OBCAM augmented synapse formation.
- Neuronal activity promoted OBCAM internalization via a raft-dependent pathway.
Conclusions:
- OBCAM functions as a synaptic cell adhesion molecule involved in synaptogenesis.
- OBCAM's surface localization is dynamically regulated by neuronal activity.
- These findings highlight OBCAM's importance in synaptic plasticity and neuronal network formation.
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