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Counting method of hair cells in Corti's organ

AuthorsSanz, Lorena ; Avendaño, Carlos; Murillo-Cuesta, Silvia ; Martínez-Vega, Raquel ; Sistiaga, J. A.; García-Alcántara, F. ; Varela-Nieto, Isabel ; Rivera, Teresa
Issue Date2012
CitationIEB (2012)
Abstract[Introduction and Objective]: The cochleogram is a graphic record which represents hair cells along the length of the basilar membrane and relates cell damage with frequency specific values in hearing thresholds. The purpose of this study is to design a simple and robust method to quantitatively determine the distribution of the inner and outer hair cells at the organ of Corti in the mouse cochlea. [Materials and Methods]: Six male CBA/CaOlaHsd mice with normal auditory brainstem responses were sacrificed at 2months of age. The cochleae from both ears (n=12) were extracted, fixed and decalcified, and then divided in two parts (apical-middle and basal), obtaining around 80% of the whole extent of the basilar membrane. The organ of Corti (OC) was isolated and phalloidin-stained in multiwall glass slides. Using a fluorescence microscope and stereological software, the total length of the OC was divided into equidistant 5% sectors1. The number of inner (IHC) and outer (OHC) hair cells in randomly distributed areas were determined, and cell density (cells/mm2) was estimated for each sector. [Results]: The distribution of hair cells along the apical, middle and available basal turns of the cochlea was fairly homogeneous. The overall mean density for IHC and OHC was similar in both sides (Left IHC: 4.10, OHC: 12.12; Right IHC: 4.20, OHC: 12.22 (p>0.05), with a ratio IHC/OHC of 2.95 and 2.91 (right and left sides, respectively), without significant differences between them, and similar to physiological 3:1 ratio. [Conclusion]: This method allows us to quantify the hair cell populations throughout the cochlear turns in a precise and reproducible manner and can be used to assess hair cell damage or regeneration in experimental mice models.
DescriptionTrabajo presentado al 49th Workshop on Inner Ear Biology celebrada en Alemania del 30 de septiembre al 2 de octubre de 2012.
Appears in Collections:(IIBM) Comunicaciones congresos
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