Presentation Details
Application of Expansion Microscopy to Study Taste

Kang-Hoon Kim, Emma Larsson, Minliang Zhou, Hong Wang.

Monell Chemical Senses Center, Philadelphia, PA, USA

Abstract


The sense of taste can distinguish at least five principal sensory qualities: salt, sweet, sour, bitter, and umami through specialized epithelial cells of taste buds, called taste receptor cells. In general, taste receptor cells express molecular receptors for only one taste quality. However, the responses of taste receptor cells are not always restricted to a single quality. In addition, responses of individual gustatory nerve fibers range from specific, responding to a single taste quality, to multimodal, responding to multiple qualities. However, the origin of this broader responsiveness is still unclear and may reside in the lack of specificity in synapses between taste cells and gustatory nerve fibers. To understand the complex subsynaptic organization of proteins in the pre- and post-synaptic terminals in the surrounding synaptic cleft, we applied hydrogel-based tissue expansion, the so-called expansion microscopy (ExM). Even if the high resolution of transmission electron microscopy (TEM) enables identification of cellular interactions within a taste bud, including taste cell-to-nerve fiber synapses as well as contacts between taste cells themselves, TEM imaging cannot always provide reliable and precise localizations of target synaptic proteins because of technical challenges regarding labeling specificity, probe size, and sample embedding. Here, we optimize antigenicity of several type of taste receptor markers and synapse molecules and improve specificity of immunostaining in hydrogel platform of the tongue tissue. We expect that the hydrogel method can be employed for studying the mechanisms underlying the regulation of synaptic architecture between taste cells and gustatory nerve fibers and providing nanoscale molecular distribution of synaptic proteins in situ.

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