Transgenic Tg(Kcnj10-ZsGreen) Fluorescent Reporter Mice Allow Visualization of Intermediate Cells in the Stria Vascularis

Author:

Strepay Dillon1,Olszewski Rafal T.1,Nixon Sydney1,Korrapati Soumya1,Adadey Samuel1,Griffith Andrew J.2,Su Yijun3,Liu Jiamin3,Vishwasrao Harshad3,Gu Shoujun1,Saunders Thomas4,Roux Isabelle2,Hoa Michael1

Affiliation:

1. Auditory Development and Restoration Program, National Institute on Deafness and Other Communication Disorders, National Institutes of Health

2. Otolaryngology Branch, National Institute on Deafness and Other Communication Disorders, National Institutes of Health

3. Advanced Imaging and Microscopy Resource, National Institutes of Health

4. Transgenic Animal Model Core, Biomedical Research Core Facility, University of Michigan

Abstract

Abstract The stria vascularis (SV) is a stratified epithelium in the lateral wall of the mammalian cochlea, responsible for both endolymphatic ion homeostasis and generation of the endocochlear potential (EP) critical for normal hearing. The SV has three layers consisting predominantly of basal, intermediate, and marginal cells. Intermediate and marginal cells form an intricate interdigitated network of cell projections making discrimination of the cells challenging. To enable intermediate cell visualization, we engineered by BAC transgenesis, reporter mouse lines expressing ZsGreen fluorescent protein under the control of Kcnj10 promoter and regulatory sequences. Kcnj10 encodes KCNJ10 protein (also known as Kir4.1 or Kir1.2), an ATP-sensitive inwardly-rectifying potassium channel critical to EP generation, highly expressed in SV intermediate cells. In these transgenic mice, ZsGreen fluorescence mimics Kcnj10 endogenous expression in the cochlea and was detected in the intermediate cells of the SV, in the inner phalangeal cells, Hensen’s, Deiters’ and pillar cells, in a subset of spiral ganglion neurons, and in glial cells. We show that expression of the transgene in hemizygous mice does not alter auditory function, nor EP. These transgenic Tg(Kcnj10-ZsGreen) mice allow live and fixed tissue visualization of ZsGreen-expressing intermediate cells and will facilitate future studies of stria vascularis cell function.

Publisher

Research Square Platform LLC

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