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Developmental switch from GABA to glycine release in single central synaptic terminals

Nature Neuroscience volume 7, pages 17–23 (2004)Cite this article

Abstract

Early in postnatal development, inhibitory inputs to rat lateral superior olive (LSO) neurons change from releasing predominantly GABA to releasing predominantly glycine into the synapse. Here we show that spontaneous miniature inhibitory postsynaptic currents (mIPSCs) also change from GABAergic to glycinergic over the first two postnatal weeks. Many 'mixed' mIPSCs, resulting from co-release of glycine and GABA from the same vesicles, are seen during this transition. Immunohistochemistry showed that a large number of terminals contained both GABA and glycine at postnatal day 8 (P8). By P14, both the content of GABA in these mixed terminals and the contribution of GABA to the mixed mIPSCs had decreased. The content of glycine in terminals increased over the same period. Our results indicate that switching from GABAergic to glycinergic inputs to the LSO may occur at the level of a single presynaptic terminal. This demonstrates a new form of developmental plasticity at the level of a single central synapse.

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Figure 1: Developmental change in the bicuculline sensitivity of IPSCs recorded in LSO neurons in response to electrical stimulation of the ventromedial aspect of the LSO brain slice.
Figure 2: Pharmacological and kinetic isolation of GABAergic, glycinergic and mixed mIPSCs in voltage-clamped, isolated LSO neurons.
Figure 3: Developmental change in mIPSCs recorded in isolated LSO neurons.
Figure 4: Developmental decrease in the contribution of the GABAergic component to mixed mIPCSs.
Figure 5: Quantitative analysis of GABA and glycine content in presynaptic terminals using immunogold staining and electron microscopy.
Figure 6: Immunohistochemical staining of GAD and glycine in the developing LSO.

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Acknowledgements

We thank A. Moorhouse for discussion and editing of the manuscript, and N. Akaike for technical advice. We also thank O.P. Otterson, I.J. Kopin, W.H. Oertel, D.E. Schmechel and M.L. Tappaz for help obtaining antibodies. This work was supported by research grants from the Ministry of Education, Culture, Sports, Science and Technology, Japan (15016082, 15650076 and 15390065 to J.N).

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Author notes

  1. Junichi Nabekura and Shutaro Katsurabayashi: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cellular and System Physiology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, 812-8582, Japan

    Junichi Nabekura, Shutaro Katsurabayashi, Yasuhiro Kakazu, Shumei Shibata, Yoshito Mizoguchi & Hitoshi Ishibashi

  2. Department of Developmental Physiology, National Institute for Physiological Sciences, Okazaki, 444-8585, Japan

    Junichi Nabekura

  3. Department of Otorhinolaryngology, School of Medicine, Hirosaki University, Hirosaki, 036-8562, Japan

    Atsushi Matsubara & Akira Sasaki

  4. Department of Anatomy and Neurobiology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, 812-8582, Japan

    Shozo Jinno

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Correspondence to Junichi Nabekura.

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Supplementary information

Supplementary Fig. 1

Sensitivity of antibody to GABA and glycine. Test sections were incubated in the same drops of glycine and GABA antibodies together with LSO sections. Note that the glycine conjugates are stained selectively positive for the 10 nm gold particles while the GABA conjugates are immunolabeled for 20 nm gold particles. None: without addition of any amino acid. (JPG 14 kb)

Supplementary Fig. 2

Histogram showing the particle densities representing the sum of GABA and glycine in the presynaptic terminals in the P14 rat LSO. Note that the histogram displays two peaks, less than 5 and more than 10. Thus, particle densities less than 5 (particles/μm2) of GABA and glycine were considered to show background level labeling. Terminals > 5 (particles/μm2) were employed for analysis as inhibitory terminals. (JPG 27 kb)

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Nabekura, J., Katsurabayashi, S., Kakazu, Y. et al. Developmental switch from GABA to glycine release in single central synaptic terminals. Nat Neurosci 7, 17–23 (2004). https://doi.org/10.1038/nn1170

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