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Volume 17, Number 19, Issue of October 1, 1997 pp. 7339-7350
Copyright ©1997 Society for Neuroscience

The Development of Excitatory Synapses in Cultured Spinal Neurons

Richard J. O'Brien^{1, 2, 3}, Andrew L. Mammen^{1, 2}, Seth Blackshaw², Michael D. Ehlers^{1, 2}, Jeffrey D. Rothstein³, and Richard L. Huganir^{1, 2}

¹ Howard Hughes Medical Institute, and Departments of ² Neuroscience and ³ Neurology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205

Immunohistochemical studies of synapses in the CNS have demonstrated that glutamate receptors (GluRs) are concentrated at postsynaptic sites in vivo and in vitro (). The mechanisms leading to receptor clustering at excitatory synapses are far less understood than those governing acetylcholine receptor accumulation at the neuromuscular junction () or glycine receptor aggregation at central inhibitory synapses (). Using cultured rat spinal cord neurons, we demonstrate that clustering of the AMPA receptor subunit GluR1 is among the earliest events in excitatory synapse formation in vitro, coincident with the onset of miniature EPSCs and in many cases preceding presynaptic vesicle accumulation. Postsynaptic receptor clustering is induced in a highly specific and reiterative pattern, independent of receptor activation, by contact with a subset of axons capable of inducing receptor clusters. The subunit composition of AMPA receptor clusters varied significantly between neurons but was invariant within a given neuron. The presence of either GluR2 or GluR3 was common to all receptor clusters. Neither high-affinity glutamate transporters nor NMDA receptors appeared to be concentrated with AMPA receptor subunits at these excitatory synapses.

Key words: GluR1; synaptogenesis; spinal cord; glutamate transporter; glutamate receptors; rat; tissue culture

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