Protocols in Current Issue
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0 Q&A 2460 Views Jan 5, 2022

Spiral ganglion neurons (SGN) are the primary neuronal pathway for transmitting sensory information from the inner ear to the brainstem. Recent studies have revealed significant biophysical and molecular diversity indicating that auditory neurons are comprised of sub-groups whose intrinsic properties contribute to their diverse functions. Previous approaches for studying the intrinsic biophysical properties of spiral ganglion neurons relied on patch-clamp and molecular analysis of cultured somata that were disconnected from their pre-synaptic hair cell partners. In the absence of the information provided by cell-to-cell connectivity, such studies could not associate biophysical diversity with functional sub-groups. Here we describe a protocol for preparing, recording, and labeling spiral ganglion neurons in a semi-intact ex-vivo preparation. In these preparations, the cell bodies of spiral ganglion neurons remain connected to their hair cell partners. The recordings are completed within 4 hours of euthanasia, alleviating concerns about whether long culture times and culture conditions change the intrinsic properties of neurons.

0 Q&A 2648 Views Dec 5, 2020

The inferior colliculus (IC) is a critical midbrain integration center for auditory and non-auditory information. Although much is known about the response properties of the IC neurons to auditory stimuli, how the IC neural circuits function during movement such as locomotion remains poorly understood. Mice offer a valuable model in this respect, but previous studies of the mouse IC were performed in anesthetized or restrained preparations, making it difficult to study the IC function during behavior. Here we describe a neural recording protocol for the mouse IC in which mice are head-fixed, but can run on a passive treadmill. Mice first receive a headpost surgery, and become habituated to head-fixing while being on a treadmill. Following a few days of habituation, neural recordings of the IC neuron activity are performed. The neural activity can be compared across different behavioral conditions, such as standing still versus running on a treadmill. We describe how to overcome the challenges of headpost surgery for awake IC recording, presented by the location and overlying bones. This protocol allows investigations of the IC function in behaving mice, while allowing precise stimulus control and the use of recording methods similar to those for anesthetized preparations.

0 Q&A 4470 Views Dec 5, 2019
Hearing loss is a common sensory deficiency suffered by millions worldwide. It is a heterogeneous condition and genetics plays a critical role in its etiology. Gene variants can fundamentally alter hearing function, or predispose the auditory system towards loss of function resulting from other factors. In mouse studies of hearing loss and gene function, an evoked potential electrophysiological recording, the auditory brainstem response (ABR), is now considered the optimal way to screen large numbers of individuals, either with normal hearing sensitivity or with hearing impairment. Other routinely used methods to assess hearing function (such as acoustic startle responses, or otoacoustic emissions) do not allow assessment of the same broad spectrum of dysfunction nor readily allow the threshold sensitivity of the neural output of the cochlea to be assessed and are less ideal. An optimized recording system to rapidly and reproducibly record high-quality ABRs from mutant mice as part of a high-throughput phenotyping pipeline was developed. Click-evoked ABRs and ABRs evoked by pure-tone frequencies over a range of sound levels from 0 dB to 95 dB, sound pressure levels (SPL) are recorded. This takes approximately 15-20 min per mouse (with 5 tone frequencies), allowing a large number of mutant mice to be screened. This method has been used to measure ABRs on a high-throughput mutant mouse phenotyping pipeline and in laboratory tests to follow-up the hearing loss phenotypes identified on that pipeline.

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