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. 2018 Feb 22;12:43. doi: 10.3389/fncel.2018.00043

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Copyright © 2018 Rosskothen-Kuhl, Hildebrandt, Birkenhäger and Illing.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Identity and neighborhood of Fos-positive cells in the CIC of deafened rats by 1 (B–F) or 7 d EIS (G–I), and of hearing rats by 1 d EIS (J–L) in confocal micrographs. (A) Whenever a collicular neuron contained a Fos-positive nucleus (arrows), it was located in HuC-/HuD-positive cytoplasm indicative of nerve cells. (B) Fos-positive nuclei (arrows) were never found in S100 positive cell bodies (some indicated by arrowheads) indicative of astrocytic cell bodies. (C) Fos-positive nuclei (arrow) were never found in IBA1 positive cell bodies indicative of microglia (some indicated by arrowheads). (D) Fos-positive cells (one indicated by arrow) were embedded in a meshwork of astrocytic processes that shared the neighborhood and showed distinct approximations (arrow-pairs). (E) Fos-positive cells were embedded in a dense population of microglia processes that showed specific spatial neuron–glia appositions (arrow-pairs). (F) The aggregations of glial processes were particularly dense where neurons showed strong Fos immunoreactivity and again showed many incidences of approximation between astrocytic processes and Fos-positive nuclei. (G) By 7 d EIS, Fos staining was waning and astrocyte aggregations have slightly eased. (H) By that time, microglia are still prominent but their acanthoid appearance seen by 1 d EIS (E) is no longer obvious. (I) At lower magnification a moderate decrease of glia aggregations can be recognized and approximations of glia processes and neurons recognizable by Fos staining are less evident by 7 d EIS compared to 1 d EIS (F). (J) When nh rats were stimulated for 1 d astrocytic processes were sparse and approximations with Fos-positive neurons were rare. (K) Upon 1 d EIS, hearing rats did not change microglia morphology against controls, leaving them with thin long extensions that rarely touch Fos-positive cell bodies. (L) Sparse astrocytic extensions, thin microglial dendrites, and less frequent Fos-positive neurons provide few opportunities of approximations between them. Scale bar for (A–C,F,I): 50 μm, scale bar for (D,E,G,H,J,K,L): 20 μm.