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  • Patch-clamp Recordings and Single Fiber Labeling from Spiral Ganglion Somata in Acutely Prepared Semi-intact Cochleae from Neonatal Rats.

Patch-clamp Recordings and Single Fiber Labeling from Spiral Ganglion Somata in Acutely Prepared Semi-intact Cochleae from Neonatal Rats.

Bio-protocol (2022-02-05)
Alexander L Markowitz, Megana R Iyer, Radha Kalluri
摘要

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.

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HEPES, ≥99.5% (titration)
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胶原酶 来源于溶组织梭菌, suitable for release of physiologically active rat hepatocytes, Type IV, 0.5-5.0 FALGPA units/mg solid, ≥125 CDU/mg solid
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胰蛋白酶 来源于牛胰腺, TPCK Treated, essentially salt-free, lyophilized powder, ≥10,000 BAEE units/mg protein
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腺苷 3',5'-环单磷酸 钠盐 一水合物, ≥98.0% (HPLC), powder
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5′-三磷酸鸟苷三钠三磷酸鸟苷三钠 锂盐, ~95% (HPLC), powder
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抗外周蛋白抗体, serum, Chemicon®