Presynaptic calcium channels and α3-integrins are complexed with synaptic cleft laminins, cytoskeletal elements and active zone components.
At chemical synapses, synaptic cleft components interact with elements of the nerve terminal membrane to promote differentiation and regulate function. Laminins containing the β2 subunit are key cleft components, and they act in part by binding the pore-forming subunit of a pre-synaptic voltage-gated calcium channel (Ca(v)α) (Nishimune et al. 2004). In this study, we identify Ca(v)α-associated intracellular proteins that may couple channel-anchoring to assembly or stabilization of neurotransmitter release sites called active zones. Using Ca(v)α-antibodies, we isolated a protein complex from Torpedo electric organ synapses, which resemble neuromuscular junctions but are easier to isolate in bulk. We identified 10 components of the complex: six cytoskeletal proteins (α2/β2 spectrins, plectin 1, AHNAK/desmoyokin, dystrophin, and myosin 1), two active zone components (bassoon and piccolo), synaptic laminin, and a calcium channel β subunit. Immunocytochemistry confirmed these proteins in electric organ synapses, and PCR analysis revealed their expression by developing mammalian motor neurons. Finally, we show that synaptic laminins also interact with pre-synaptic integrins containing the α3 subunit. Together with our previous finding that a distinct synaptic laminin interacts with SV2 on nerve terminals (Son et al. 2000), our results identify three paths by which synaptic cleft laminins can send developmentally important signals to nerve terminals.
Carlson
Steven S
SS
Department of Physiology and Biophysics, University of Washington, Seattle, Washington 98195, USA. ssc1@u.washington.edu
Valdez
Gregorio
G
Sanes
Joshua R
JR
D000818:Animals; D001369:Axons / Q000378:metabolism; D015220:Calcium Channels / Q000378:metabolism*; D051242:Caveolin 1 / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D039422:Integrin alpha3 / Q000378:metabolism*; D007797:Laminin / Q000378:metabolism*; D051379:Mice; D009046:Motor Neurons / Q000378:metabolism; D009411:Nerve Endings / Q000378:metabolism; D011485:Protein Binding; D011817:Rabbits; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013569:Synapses / Q000378:metabolism*; D014101:Torpedo / Q000502:physiology
D015220:Calcium Channels; D051242:Caveolin 1; D003598:Cytoskeletal Proteins; D039422:Integrin alpha3; D007797:Laminin