Expression of synapsin I correlates with maturation of the neuromuscular synapse.
Synapsins are a family of neuron-specific phosphoproteins that are localized within the presynaptic terminals in adult brain. Previous work has demonstrated that introduction of exogenous synapsins I(a + b) or IIa into Xenopus spinal neurons promoted maturation of the neuromuscular synapse in a nerve-muscle co-culture system. We have now studied the expression of endogenous Xenopus synapsin I during synaptic maturation in vivo and in culture, using a polyclonal antibody raised against Xenopus synapsin I. Immunoprecipitation experiments indicated that synapsin I was not detectable during the early phase of synaptogenesis in vivo, and exhibited a marked increase during the period of synaptic maturation. In contrast, the expression of synaptophysin, another synaptic vesicle protein, was detected at the start of nervous system formation, and remained at a high level thereafter. Similar expression profiles for the two proteins were also observed in immunocytochemical studies of Xenopus spinal neurons in culture: intense staining of synaptophysin was found on the first day, while synapsin I was not detected until after three days in culture. The expression of synapsin I correlated very well with the appearance of a bell-shaped amplitude distribution of spontaneous synaptic currents, a physiological parameter which reflects functional maturation of the neuromuscular synapse. In one-day-old cultures grown in the absence of laminin, an extracellular matrix protein known to be present at the neuromuscular junction, the amplitude distribution of virtually all synapses was skewed towards smaller values. In contrast, when laminin was used as a culture substrate, many synapses exhibited a bell-shaped amplitude distribution. Laminin treatment also induced synapsin I expression in one-day-old cultures. These results suggest that the expression of endogenous synapsin I may regulate maturation at neuromuscular synapses.
Laboratory of Molecular and Cellular Neuroscience, Rockefeller University, New York 10021-6399, USA.
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Popov
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Wang
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Greengard
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D018920:Coculture Techniques; D004594:Electrophysiology; D007150:Immunohistochemistry; D007797:Laminin / Q000494:pharmacology; D008564:Membrane Potentials / Q000502:physiology; D024510:Muscle Development; D018482:Muscle, Skeletal / Q000254:growth & development* / Q000294:innervation / Q000502:physiology; D009469:Neuromuscular Junction / Q000502:physiology*; D018408:Patch-Clamp Techniques; D011233:Precipitin Tests; D013569:Synapses / Q000378:metabolism / Q000502:physiology*; D016704:Synapsins / Q000096:biosynthesis*; D016708:Synaptophysin / Q000096:biosynthesis; D014982:Xenopus laevis
D007797:Laminin; D016704:Synapsins; D016708:Synaptophysin