The pattern of expression of ionotropic glutamate receptor (GluR) subunits 1-4 in the medial nucleus of the trapezoid body (MNTB) has been reported to change during development. The aim of this study was to compare the distribution of the GluR1-4 subunits in the MNTB at postnatal day (P) 9, before high-frequency signal transmission in the auditory system has developed, with that observed in mature adult rats. GluR1-4 subunits were studied by preembedding and postembedding immunocytochemical methods. Increased levels of GluR1, 2/3, and 4 associated with development were evident only at postsynaptic sites of MNTB principal cell bodies receiving calyces of Held synapses, whereas receptor density at nonsynaptic sites was found to remain unaltered. Taken together, the expression pattern of GluR subunits and the density of immunoparticles in postsynaptic specializations are indicative of a compartmentalization of alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) subunits upon development. These developmental changes provide a morphological basis for establishment of the postsynaptic properties needed for high-frequency synaptic transmission of auditory signals to MNTB principal neurons.
The mature calyx of Held ending on principal neurons of the medial nucleus of the trapezoid body (MNTB) has very specialized morphological and molecular features that make it possible to transmit auditory signals with high fidelity. In a previous work we described an increased localization of the ionotropic alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) glutamate receptor (GluA) subunits at postsynaptic sites of the calyx of Held-principal cell body synapses from postnatal development to adult. The aim of the present study was to investigate whether the pattern of the synaptic distribution of GluA2/3/4c and -4 in adult MNTB principal cell bodies correlated with preferential subcellular domains (stalks and swellings) of the calyx. We used a postembedding immunocytochemical method combined with specific antibodies to GluA2/3/4c and GluA4 subunits. We found that the density of GluA2/3/4c in calyceal swellings (19 +/- 1.54 particles/microm) was higher than in stalks (10.93 +/- 1.37 particles/microm); however, the differences for GluA4 were not statistically significant (swellings: 13.84 +/- 1.39 particles/microm; stalks: 10.42 +/- 1.24 particles/microm). Furthermore, GluA2/3/4c and GluA4 labeling co-localized to some extent in calyceal stalks and swellings. Taking these data together, the distribution pattern of GluA subunits in postsynaptic specializations are indicative of a spatial compartmentalization of AMPA subunits in mature calyx-principal neuron synapses that may support the temporally precise transmission required for sound localization in the auditory brainstem.
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