Elsevier

Neuroscience

Volume 178, 31 March 2011, Pages 218-239
Neuroscience

Pain Mechanisms and Sensory Neuroscience
Research Paper
Vesicular glutamate transporters: spatio-temporal plasticity following hearing loss

https://doi.org/10.1016/j.neuroscience.2010.12.059Get rights and content

Abstract

An immunocytochemical comparison of vGluT1 and vGluT3 in the cochlear nucleus (CN) of deafened versus normal hearing rats showed the first example of vGluT3 immunostaining in the dorsal and ventral CN and revealed temporal and spatial changes in vGluT1 localization in the CN after cochlear injury. In normal hearing rats vGluT1 immunostaining was restricted to terminals on CN neurons while vGluT3 immunolabeled the somata of the neurons. This changed in the ventral cochlear nucleus (VCN) 3 days following deafness, where vGluT1 immunostaining was no longer seen in large auditory nerve terminals but was instead found in somata of VCN neurons. In the dorsal cochlear nucleus (DCN), while vGluT1 labeling of terminals decreased, there was no labeling of neuronal somata. Therefore, loss of peripheral excitatory input results in co-localization of vGluT1 and vGluT3 in VCN neuronal somata. Postsynaptic glutamatergic neurons can use retrograde signaling to control their presynaptic inputs and these results suggest vGluTs could play a role in regulating retrograde signaling in the CN under different conditions of excitatory input. Changes in vGluT gene expression in CN neurons were found 3 weeks following deafness using qRT-PCR with significant increases in vGluT1 gene expression in both ventral and dorsal CN while vGluT3 gene expression decreased in VCN but increased in DCN.

Research highlights

▶vGluT3 is expressed and produced in the cochlear nucleus. ▶Activity level governs neuronal compartmentalization of vGluTs. ▶Plasticity dependent co-localization of vGluTs occurs after hearing loss. ▶Activity dependent somatic localization of vGluTs suggests retrograde signaling.

Section snippets

Experimental procedures

All experimental procedures were approved by the Institutional Animal Care and Use Committee at Wayne State University and conform to NIH guidelines. Adult, male Sprague–Dawley rats (Charles River Laboratories, Wilmington, MA, USA) were used in this study.

Cochlear ablation results in profound hearing loss

To determine the hearing status of each of the animals, auditory brainstem responses were analyzed prior to and after cochlear hair cell ablation. Normal hearing animals showed average thresholds of 30 dB at 4 kHz, 28 dB at 12 kHz, and 30 dB at 20 kHz. In no case did any animal have detectable thresholds (up to 100 dB) at 3 days, 3 weeks, or 2 months following cochlear hair cell ablation (Fig. 1A) at any frequency tested. The absence of ABR thresholds following cochlear hair cell ablation

Discussion

In the current study we report the distribution of vGluT3 in the CN. We also demonstrate spatial and temporal changes in gene expression for all three vGluTs as a consequence of deafness. Previous studies of other systems support the idea of a complementary distribution of vGluTs (one neuron produces one vGluT) with vGluT1 reported primarily in cortex, hippocampus, and cerebellar cortex, vGluT2 reported in neurons of the thalamus and brainstem and vGluT3 reported in the caudate-putamen, the

Acknowledgments

These studies were supported by R03 NIDCD Grant DC007733 to AGH, and P30 Grants (Anatomy and Cell Biology, Wayne State University; Kresge Hearing Research Institute, University of MI). We would like to acknowledge the technical assistance of K. Vistisen with Western blot and Dr. R. Armant for assistance with Agilent analysis. We thank Dr. J. Erickson for providing control peptides for the mfusion-vGluT antibodies and Dr. M. Drescher for critical comments about the Organ of Corti analysis. We

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