Ontogeny of cellular organization and LGR5 expression in porcine cochlea revealed using tissue clearing and 3D imaging

Adele Moatti; Chen Li; Sasank Sivadanam; Yuheng Cai; James Ranta; Jorge A Piedrahita; Alan G Cheng; Frances S Ligler; Alon Greenbaum

doi:10.1016/j.isci.2022.104695

Ontogeny of cellular organization and LGR5 expression in porcine cochlea revealed using tissue clearing and 3D imaging

iScience. 2022 Jun 30;25(8):104695. doi: 10.1016/j.isci.2022.104695. eCollection 2022 Aug 19.

Authors

Adele Moatti^{1

2}, Chen Li^{1

2}, Sasank Sivadanam¹, Yuheng Cai^{1

2}, James Ranta³, Jorge A Piedrahita^{1

2}, Alan G Cheng⁴, Frances S Ligler^{1

2}, Alon Greenbaum^{1

2}

Affiliations

¹ Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27606, USA.
² Comparative Medicine Institute, North Carolina State University, Raleigh, NC 27606, USA.
³ Department of Electrical Engineering, North Carolina State University, Raleigh, NC 27606, USA.
⁴ Department of Otolaryngology-Head and Neck Surgery, Stanford University School of Medicine, Stanford, CA 94305, USA.

Abstract

Over 11% of the world's population experience hearing loss. Although there are promising studies to restore hearing in rodent models, the size, ontogeny, genetics, and frequency range of hearing of most rodents' cochlea do not match that of humans. The porcine cochlea can bridge this gap as it shares many anatomical, physiological, and genetic similarities with its human counterpart. Here, we provide a detailed methodology to process and image the porcine cochlea in 3D using tissue clearing and light-sheet microscopy. The resulting 3D images can be employed to compare cochleae across different ages and conditions, investigate the ontogeny of cochlear cytoarchitecture, and produce quantitative expression maps of LGR5, a marker of cochlear progenitors in mice. These data reveal that hair cell organization, inner ear morphology, cellular cartography in the organ of Corti, and spatiotemporal expression of LGR5 are dynamic over developmental stages in a pattern not previously documented.

Keywords: Biological sciences research methodologies; Cell biology; Imaging anatomy; Optical imaging.