Whole organ imaging of the mature and aged mammalian vestibular system.

Goettl-Meyer, M.; Perez-Guevara, M.; Pendyala, P.; Caprara, G. A.; Rennie, K. J.; Peng, A. W.

Aging is associated with an increased risk of falling, which is contributed to by diminished balance. However, the precise anatomical changes occurring within the vestibular system as it ages are debated. In this study, we used immunofluorescence, tissue clearing, and 2-photon microscopy to visualize the anatomical changes that occur as the vestibular system ages. The entire temporal bone was dissected and decalcified, instead of dissecting individual vestibular organs, to minimize tissue damage and distortions during dissection. We compared tested multiple clearing methods, both aqueous and solvent based methods to find which works best for the vestibular organs and settled on using the ethyl cin-namate method. These methods allow imaging of the entire vestibular system sensory cells in their native orientations. Using a combination of antibodies, we can label all hair cells as well as the type II hair cells. Using automated analyses, we delineate the region of the sensory epithelia and automate the counting and mapping of the locations of each cell. Using this protocol, we quantify the number of hair cells in both mature (1-2 months) and aged (36-40 months) epithelia to determine how hair cell numbers change with age. We also correlate these changes with vestibular function using vestibular evoked potential recordings. This study lays the groundwork for determining changes in the vestibular system with age to determine the pathophysiological changes.

POLAND; VESTIBULAR apparatus; PUBERTY; FLUORESCENT antibody technique; CONFERENCES & conventions; MAMMALS; AGING; MICROSCOPY

Journal of Hearing Science, 2024, Vol 14, Issue 3, p119

2083-389X

Academic Journal