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The Journal of Neuroscience, February 1, 2000, 20(3):1009-1019
The Presynaptic Calcium Channel Is Part of a Transmembrane
Complex Linking a Synaptic Laminin ( 4 2 1) with
Non-Erythroid Spectrin
William J.
Sunderland1,
Young-Jin
Son1,
Jeffrey H.
Miner2,
Joshua R.
Sanes3, and
Steven S.
Carlson1
1 Department of Physiology and Biophysics, University
of Washington, Seattle, Washington 98195, and Departments of
2 Medicine and 3 Anatomy and Neurobiology,
Washington University School of Medicine, St. Louis, Missouri
63110
Nerve regeneration studies at the neuromuscular junction (NMJ)
suggest that synaptic basal lamina components tell the returning axon
where to locate neurotransmitter release machinery, including synaptic
vesicle clusters and active zones. Good candidates for these components
are the synaptic laminins (LNs) containing  4, 5, or  2 chains.
Results from a 2 laminin knockout mouse have suggested a linkage of
this extracellular laminin to cytosolic synaptic vesicle clusters. Here
we report such a transmembrane link at the electric organ synapse,
which is homologous to the NMJ. We immunopurified electric organ
synaptosomes and found on their surface two laminins of 740 and 900 kDa. The 740 kDa laminin has a composition of 42 1 (laminin-9).
Immunostaining reveals that as in the NMJ, 4 and 2 chains are
concentrated at the electric organ synapse. Using detergent-solubilized
synaptosomes, we immunoprecipitated a complex containing 421
laminin, the voltage-gated calcium channel, and the cytoskeletal
protein spectrin. Other presynaptic proteins such as 900 kDa laminin
are not found in this complex. We hypothesize that 421 laminin
in the synaptic basal lamina attaches to calcium channel, which in turn
is attached to cytosolic spectrin. Spectrin could then organize
synaptic vesicle clusters by binding vesicle-associated proteins.
Key words:
calcium channel; laminin; spectrin; electric organ; synapses; synaptosomes; nerve terminal
Copyright © 2000 Society for Neuroscience 0270-6474/00/2031009-11$05.00/0
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