Matrix metalloproteinase-3 removes agrin from synaptic basal lamina.
Agrin, a heparin sulfate proteoglycan, is an integral member of the synaptic basal lamina and plays a critical role in the formation and maintenance of the neuromuscular junction. The N-terminal region of agrin binds tightly to basal lamina, while the C-terminal region interacts with a muscle-specific tyrosine kinase (MuSK) to induce the formation of the postsynaptic apparatus. Although the binding of agrin to basal lamina is tight, the binding of agrin to MuSK has yet to be shown; therefore, basal lamina binding is critical for maintaining the presentation of agrin to MuSK. Here we report evidence that supports our hypothesis that matrix metalloproteinase-3 (MMP-3) is responsible for the removal of agrin from synaptic basal lamina. Antibodies to the hinge region of human MMP-3 recognize molecules concentrated at the frog neuromuscular junction in both cross sections and whole mounts. Electron microscopy of neuromuscular junctions stained with antibodies to MMP-3 reveals that staining is found in the extracellular matrix surrounding the Schwann cell. Treatment of sections from frog anterior tibialis muscle with MMP-3 results in a clear and reproducible removal of agrin immunoreactivity from synaptic basal lamina. The same MMP-3 treatment does not alter anti-laminin staining. These results support our hypothesis that synaptic activity results in the activation of MMP-3 at the neuromuscular junction and that MMP-3 specifically removes agrin from synaptic basal lamina.
VanSaun
M
M
Department of Anatomy and Cell Biology, The University of Kansas Medical Center, Kansas City, Kansas 66160, USA.
Werle
M J
MJ
D018171:Agrin / Q000378:metabolism*; D000818:Animals; D000906:Antibodies / Q000378:metabolism; D000918:Antibody Specificity; D001485:Basement Membrane / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D002038:Bungarotoxins / Q000493:pharmacokinetics; D004305:Dose-Response Relationship, Drug; D007150:Immunohistochemistry; D007797:Laminin / Q000378:metabolism; D008297:Male; D019278:Matrix Metalloproteinase 3 / Q000378:metabolism* / Q000494:pharmacology; D008854:Microscopy, Electron; D018482:Muscle, Skeletal / Q000187:drug effects / Q000201:enzymology / Q000294:innervation; D009469:Neuromuscular Junction / Q000378:metabolism* / Q000648:ultrastructure; D011894:Rana pipiens; D012235:Rhodamines; D013570:Synaptic Membranes / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure
D018171:Agrin; D000906:Antibodies; D002038:Bungarotoxins; D007797:Laminin; D012235:Rhodamines; D019278:Matrix Metalloproteinase 3