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In-vivo optogenetic manipulation approach for gerbil medial nucleus of trapezoid body

Ben‐Zheng LiiD, Matthew Ridenour, Shani PolegiD, Praveen KuruppathiD, Elizabeth A. McCullaghiD, Tim C. LeiiD, Achim KlugiD

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Abstract

PurposeThis study introduces an in-vivo optogenetic manipulation approach for the medial nucleus of the trapezoid body (MNTB) in Mongolian gerbils, a species with a hearing range similar to humans. The MNTB is crucial for sound localization, but traditional methods lack temporal precision and reversibility. The aim of this study is to develop a specific, reversible method for controlling MNTB activity with fast and precise temporal control, an approach vital for studying sound localization. MethodsWe stereotactically injected adeno-associated viral vectors encoding opsins into the gerbil MNTB. Precise targeting was achieved despite the MNTBs location in a deep, heavily myelinated area of the brainstem. Opsin expression was confirmed via confocal microscopy. In-vivo electrophysiology combined with optical stimulation was used to test optical activation and suppression of MNTB activity during sound stimuli. ResultsOpsin expression was strong and stable in MNTB neurons over weeks and months. Laser stimulation during in-vivo recordings successfully induced both activation and suppression of MNTB neurons, demonstrating fast and precise control over neural activity. ConclusionThis in-vivo optogenetic method provides specific, reversible control of MNTB activity in gerbils with rapid, real-time modulation. It offers a powerful tool for investigating sound localization and auditory processing with high temporal precision.

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