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Volume 17, Number 9,
Issue of May 1, 1997
pp. 2947-2958
Copyright ©1997 Society for Neuroscience
Activity-Dependent Expression of NT-3 in Muscle Cells in Culture:
Implications in the Development of Neuromuscular Junctions
Received Dec. 2, 1996; revised Jan. 23, 1997; accepted Feb. 10, 1997.
Kewei Xie,
Ti Wang,
Petur Olafsson,
Keiko Mizuno, and
Bai Lu
Laboratory of Developmental Neurobiology, National Institute of
Child Health and Human Development, National Institutes of Health,
Bethesda, Maryland 20892-4480
Although activity-dependent expression of neurotrophins has been
studied extensively in the CNS, its physiological role during synapse
development is not well established. At the developing neuromuscular
junction in culture, exogenous application of the neurotrophin BDNF or
NT-3 has been shown to acutely potentiate synaptic transmission and
chronically promote synapse maturation. Using the same cell culture
model, we have investigated activity-dependent neurotrophin expression
in muscle cells and its role in developing neuromuscular synapses.
Membrane depolarization, elicited by either depolarizing agents or
repetitive electric stimulation, rapidly and specifically increased the
levels of NT-3 mRNA in developing Xenopus laevis muscle
cells in culture. NT-3 gene expression also was enhanced by
acetylcholine (ACh), the neurotransmitter that causes muscle membrane
depolarization. The effects of depolarization were mediated by
increasing intracellular calcium concentration. Moreover, factor(s)
induced by membrane depolarization appeared to enhance synaptic
transmission at the developing neuromuscular junction. The frequency of
spontaneous synaptic currents (SSCs) recorded from neuromuscular
synapses was increased significantly after treatment with conditioned
medium from depolarized muscle cultures. The amplitude, rise time, and
decay time of SSCs were not affected, indicating a presynaptic action
of the conditioned medium. The effects of the conditioned medium were
blocked, partially, by the NT-3 scavenger TrkC-IgG, suggesting that the
potentiation of synaptic efficacy was attributable, at least in part,
to elevated NT-3 as a consequence of muscle depolarization. Thus,
activity-dependent expression of muscle NT-3 may contribute to the
development of the neuromuscular synapse.
Key words:
neurotrophins;
RT-PCR;
activity-dependent;
membrane
depolarization;
neuromuscular synapse;
nerve-muscle
co-culture
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