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First published online on August 12, 2004.
 Copyright © 2004 by The Physiological Society
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jphysiol.2004.067017v1
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Received April 23, 2004
Revised June 25, 2004
Accepted after revision August 11, 2004

Distinct re-filling profiles of inhibitory neurotransmitters into presynaptic terminals projecting to spinal neurons in immature rats

Shutaro Katsurabayashi1, Hisahiko Kubota2, Hideho Higashi3, Norio Akaike4*, and Yushi Ito5

1 Baylor College of Medicine
2 Hamamatsu University School of Medicine
3 Kurume University School of Medicine
4 Kumamoto Health Science University
5 Kyushu University

* To whom correspondence should be addressed. E-mail: akaike{at}kumamoto-hsu.ac.jp.

Co-release of glycine and GABA from the single synaptic terminal (synaptic bouton) is well accepted in immature rat spinal cord and brainstem. However, it raises the question of how glycine and GABA are accumulated in the same synaptic vesicles and co-released. To address this issue, spontaneous miniature inhibitory postsynaptic currents (mIPSCs) and focally evoked IPSCs (eIPSCs) mediated via a single synapse were recorded from synaptic bouton preparations of the rat immature sacral dorsal commissural nucleus (SDCN) neurons by whole-cell patch recording. Focal stimulation of a single synaptic bouton revealed that three different quantal releases occur from a single synaptic bouton: i.e., pure glycine, pure GABA, and mixed. Prolonged treatment with bafilomycin A1, a vacuolar-type H+/ATPase inhibitor, to the SDCN neuron greatly suppressed frequency and amplitude of the mIPSCs. During washing out of bafilomycin A1, complete recovery in the amplitude of glycinergic mIPSCs was observed, while that of GABAergic and mixed mIPSCs was incomplete. These observations indicate that three types of vesicles co- exist in single synaptic terminals, and that re-filling of glycine into the synaptic vesicle occurs to GABA, after the pretreatment with bafilomycin A1 in immature rats. This could be explained by the decrease in the cytosolic concentration of GABA, or by the presence of subtypes of vesicular inhibitory amino acid transporter in the synaptic vesicle membrane.


Key words: GABA • Glycine • Presynaptic terminal




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