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The Journal of Neuroscience, January 26, 2005, 25(4):799-807; doi:10.1523/JNEUROSCI.4256-04.2005

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Cellular/Molecular
Number and Density of AMPA Receptors in Single Synapses in Immature Cerebellum

Jun-ichi Tanaka,^1,2 Masanori Matsuzaki,^1,3 Etsuko Tarusawa,^1,2 Akiko Momiyama,² Elek Molnar,⁵ Haruo Kasai,^1,3 and Ryuichi Shigemoto^1,2,4

¹Department of Physiological Sciences, Graduate University for Advanced Studies, Sokendai 444-8787, Japan, Divisions of ²Cerebral Structure and ³Cell Physiology, National Institute for Physiological Sciences, Myodaiji, Okazaki 444-8787, Japan, ⁴Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, Kawaguchi, Saitama 332-0012, Japan, and ⁵Medical Research Council Centre for Synaptic Plasticity, Department of Anatomy, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, United Kingdom

The number of ionotropic receptors in synapses is an essential factor for determining the efficacy of fast transmission. We estimated the number of functional AMPA receptors at single postsynaptic sites by a combination of two-photon uncaging of glutamate and the nonstationary fluctuation analysis in immature rat Purkinje cells (PCs), which receive a single type of excitatory input from climbing fibers. Areas of postsynaptic membrane specialization at the recorded synapses were measured by reconstruction of serial ultrathin sections. The number of functional AMPA receptors was proportional to the synaptic area with a density of 1280 receptors/µm². Moreover, highly sensitive freeze-fracture replica labeling revealed a homogeneous density of immunogold particles for AMPA receptors in synaptic sites (910 ± 36 particles/µm²) and much lower density in extrasynaptic sites (19 ± 2 particles/µm²) in the immature PCs. Our results indicate that in this developing synapse, the efficacy of transmission is determined by the synaptic area.

Key words: AMPA; cerebellum; freeze fracture; postnatal; Purkinje cell; receptor; slice; synapse; ultrastructure; glutamate

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Received Oct 13, 2004; revised November 30, 2004; accepted December 2, 2004.

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