Longitudinal imaging of the taste bud in vivo with two-photon laser scanning microscopy.

Taste bud cells in the tongue transduce taste information from chemicals in food and transmit this information to gustatory neurons in the geniculate ganglion that innervate taste buds. The peripheral taste system is a dynamic environment where taste bud cells are continuously replaced, but further...

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Main Authors: Brittany N Walters, Zachary D Whiddon, Aaron W McGee, Robin F Krimm
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2024-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0309366
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author Brittany N Walters
Zachary D Whiddon
Aaron W McGee
Robin F Krimm
author_facet Brittany N Walters
Zachary D Whiddon
Aaron W McGee
Robin F Krimm
author_sort Brittany N Walters
collection DOAJ
description Taste bud cells in the tongue transduce taste information from chemicals in food and transmit this information to gustatory neurons in the geniculate ganglion that innervate taste buds. The peripheral taste system is a dynamic environment where taste bud cells are continuously replaced, but further understanding of this phenomenon has been limited by the inability to directly observe this process. To overcome this challenge, we combined chronic in vivo two-photon laser scanning microscopy with genetic labeling of gustatory neurons and taste buds to observe how cells within the taste bud change over time. This method expands the investigative possibilities beyond those offered by fixed-tissue methods. This method permits direct observation of taste bud cell entry, cell differentiation, cell loss, and arbor plasticity. We demonstrate that a few stains/dyes can be used to observe nuclei and organelles in the taste bud in vivo. We also describe a workflow for reconstructing composite z-stacks with grayscale data of both cells and arbors using ImageJ, Neurolucida 360, and Neurolucida Explorer software. Together, the methodology and software options for analyses presented here provide a novel approach for longitudinally observing taste bud cells and arbors in the taste bud in vivo.
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spelling doaj-art-1f968dbbbb6e4d58a3aea0728ce5b4f52025-08-20T02:59:45ZengPublic Library of Science (PLoS)PLoS ONE1932-62032024-01-011912e030936610.1371/journal.pone.0309366Longitudinal imaging of the taste bud in vivo with two-photon laser scanning microscopy.Brittany N WaltersZachary D WhiddonAaron W McGeeRobin F KrimmTaste bud cells in the tongue transduce taste information from chemicals in food and transmit this information to gustatory neurons in the geniculate ganglion that innervate taste buds. The peripheral taste system is a dynamic environment where taste bud cells are continuously replaced, but further understanding of this phenomenon has been limited by the inability to directly observe this process. To overcome this challenge, we combined chronic in vivo two-photon laser scanning microscopy with genetic labeling of gustatory neurons and taste buds to observe how cells within the taste bud change over time. This method expands the investigative possibilities beyond those offered by fixed-tissue methods. This method permits direct observation of taste bud cell entry, cell differentiation, cell loss, and arbor plasticity. We demonstrate that a few stains/dyes can be used to observe nuclei and organelles in the taste bud in vivo. We also describe a workflow for reconstructing composite z-stacks with grayscale data of both cells and arbors using ImageJ, Neurolucida 360, and Neurolucida Explorer software. Together, the methodology and software options for analyses presented here provide a novel approach for longitudinally observing taste bud cells and arbors in the taste bud in vivo.https://doi.org/10.1371/journal.pone.0309366
spellingShingle Brittany N Walters
Zachary D Whiddon
Aaron W McGee
Robin F Krimm
Longitudinal imaging of the taste bud in vivo with two-photon laser scanning microscopy.
PLoS ONE
title Longitudinal imaging of the taste bud in vivo with two-photon laser scanning microscopy.
title_full Longitudinal imaging of the taste bud in vivo with two-photon laser scanning microscopy.
title_fullStr Longitudinal imaging of the taste bud in vivo with two-photon laser scanning microscopy.
title_full_unstemmed Longitudinal imaging of the taste bud in vivo with two-photon laser scanning microscopy.
title_short Longitudinal imaging of the taste bud in vivo with two-photon laser scanning microscopy.
title_sort longitudinal imaging of the taste bud in vivo with two photon laser scanning microscopy
url https://doi.org/10.1371/journal.pone.0309366
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