Volume gradients in inner hair cell-auditory nerve fiber pre- and postsynaptic proteins differ across mouse strains

In different animal models, auditory nerve fibers display variation in spontaneous activity and response threshold. Functional and structural differences among inner hair cell ribbon synapses are believed to contribute to this variation. The relative volumes of synaptic proteins at individual synapses might be one such difference. This idea is based on the observation of opposing volume gradients of the presynaptic ribbons and associated postsynaptic glutamate receptor patches in mice along the pillar modiolar axis of the inner hair cell, the same axis along which fibers were shown to vary in their physiological properties. However, it is unclear whether these opposing gradients are expressed consistently across animal models. In addition, such volume gradients observed for separate populations of presynaptic ribbons and postsynaptic glutamate receptor patches suggest different relative volumes of these synaptic structures at individual synapses; however, these differences have not been examined in mice. Furthermore, it is unclear whether such gradients are limited to these synaptic proteins. Therefore, we analyzed organs of Corti isolated from CBA/CaJ, C57BL/6, and FVB/NJ mice using immunofluorescence, confocal microscopy, and quantitative image analysis. We find consistent expression of presynaptic volume gradients across strains of mice and inconsistent expression of postsynaptic volume gradients. We find differences in the relative volume of synaptic proteins, but these are different between CBA/CaJ mice, and C57BL/6 and FVB/NJ mice. We find similar results in C57BL/6 and FVB/NJ mice when using other postsynaptic density proteins (Shank1, Homer, and PSD95). These results have implications for the mechanisms by which volumes of synaptic proteins contribute to variations in the physiology of individual auditory nerve fibers and their vulnerability to excitotoxicity

Changes in spontaneous movement in response to silent gaps are not robust enough to indicate the perception of tinnitus in mice

Approaches to identify the perception of tinnitus in various animal models have been difficult to apply to mouse. As a result, mice have been underutilized to investigate the cellular, molecular, and genetic mechanisms underlying tinnitus. A recent study in guinea pigs identified a novel spontaneous behavior (unconditioned response), changes in movement during silent gaps, that identified a subgroup of animals presumably with tinnitus. Guinea pigs identified with tinnitus failed to ''freeze'' in response to silent gaps in sound. In the hope of developing a rapid and reliable assay for mice, we used a similar approach. C57BL/6J mice underwent three trials in which spontaneous movement was video recorded in the presence of white noise interrupted with six silent gaps. Movement metrics included velocity and body movement. Before the third trial, mice underwent either sham or noise exposure to induce hearing loss and tinnitus. Auditory brainstem responses before and after noise trauma confirmed normal hearing in sham-treated animals and hearing loss in the noise-exposed cohort. No differences in the various movement metrics were detected during the silent gaps either before or after sham/noise exposure. Variability in spontaneous movement both before and after sham/noise exposure was substantially greater in mice compared to guinea pigs. Thus, this assay is not sufficiently statistically powerful to identify changes in movement that might indicate tinnitus perception in mice. Previous observations also reported increased movement overall in guinea pigs identified as suffering tinnitus. In contrast, mice showed no statistically significant differences in movement between the three trials. Despite our results, other unconditioned (as well as conditioned) behaviors should be examined in mice to test their utility to detect changes that indicate the perception of tinnitus. Such assays are essential to accelerate the use of mouse models in tinnitus research