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
Affiliations
Adele Moatti
Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27606, USA; Comparative Medicine Institute, North Carolina State University, Raleigh, NC 27606, USA
Chen Li
Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27606, USA; Comparative Medicine Institute, North Carolina State University, Raleigh, NC 27606, USA
Sasank Sivadanam
Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27606, USA
Yuheng Cai
Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27606, USA; Comparative Medicine Institute, North Carolina State University, Raleigh, NC 27606, USA
James Ranta
Department of Electrical Engineering, North Carolina State University, Raleigh, NC 27606, USA
Jorge A. Piedrahita
Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27606, USA; Comparative Medicine Institute, North Carolina State University, Raleigh, NC 27606, USA
Alan G. Cheng
Department of Otolaryngology-Head and Neck Surgery, Stanford University School of Medicine, Stanford, CA 94305, USA
Frances S. Ligler
Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27606, USA; Comparative Medicine Institute, North Carolina State University, Raleigh, NC 27606, USA
Alon Greenbaum
Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27606, USA; Comparative Medicine Institute, North Carolina State University, Raleigh, NC 27606, USA; Corresponding author
Summary: 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.
