Erratic deposition of agrin during the formation of Xenopus neuromuscular junctions in culture.
In order to disclose the mechanism that regulate synapse development we compared the distributions of agrin, acetylcholine receptors (AChR) and a basal lamina heparan sulfate proteoglycan (HSPG) in sections and cultures prepared from Xenopus laevis and Ambystoma mexicanum embryos. While agrin, AChR and HSPG may accumulate almost synchronously at synapses in vivo, agrin deposition usually lagged well behind the other synaptic markers during development in culture, and was not detectable at many differentiated junctions. Agrin deposition at nerve-muscle contacts in culture also appeared to require the presence of other synaptic components. A similarly variable deposition occurred on noninnervated myocytes, where agrin again collected near sites of HSPG and AChR accumulation on some cells. Profuse agrin accretion occurred consistently, however, within the extracellular matrices of surrounding epithelial cells derived from both myotomes and neural tubes. In cocultures of Ambystoma neurons and Xenopus myocytes Ambystoma agrin collected at some chimeric neuromuscular junctions, but also accumulated on noninnervated myocytes and in the extracellular matrices of salamander neuroendothelial cells. Based upon these observations we conclude that (a) focal agrin deposition is not required for synaptic differentiation on Xenopus myocytes and (b) agrin may be one of several muscle basal lamina components that stem mainly from the secreted products of nearby epithelial cells.
Anderson
M J
MJ
Department of Anatomy, University of Calgary, Alberta, Canada.
Shi
Z Q
ZQ
Grawel
R
R
Zackson
S L
SL
D018171:Agrin / Q000032:analysis* / Q000276:immunology / Q000502:physiology; D000557:Ambystoma / Q000196:embryology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001485:Basement Membrane / Q000648:ultrastructure; D004627:Embryonic Induction; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000032:analysis*; D008856:Microscopy, Fluorescence; D009469:Neuromuscular Junction / Q000737:chemistry* / Q000196:embryology / Q000648:ultrastructure; D009924:Organ Culture Techniques; D011509:Proteoglycans / Q000032:analysis*; D011950:Receptors, Cholinergic / Q000032:analysis*; D013045:Species Specificity; D014982:Xenopus laevis / Q000196:embryology*
D018171:Agrin; D000911:Antibodies, Monoclonal; D016326:Extracellular Matrix Proteins; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D011509:Proteoglycans; D011950:Receptors, Cholinergic