A Comparison of Collection Techniques for Gene Expression Analysis of Human Oral Taste Tissue.

Nicholas Steven Archer,Jan Shaw,Konsta Duesing,Garry Hannan,Russell Keast,Dongli Liu

doi:10.1371/journal.pone.0152157

Nicholas Steven Archer, Jan Shaw + Show 4 more

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https://doi.org/10.1371/journal.pone.0152157

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Abstract

Variability in human taste perception is associated with both genetic and environmental factors. The influence of taste receptor expression on this variability is unknown, in part, due to the difficulty in obtaining human oral tissue that enables quantitative expression measures of taste genes. In a comparison of six current techniques (Oragene RNeasy Kit, Isohelix swab, Livibrush cytobrush, tongue saliva, cheek saliva collection, and fungiform papillae biopsy), we identify the fungiform papillae biopsy is the optimal sampling technique to analyse human taste gene expression. The fungiform papillae biopsy resulted in the highest RNA integrity, enabling amplification of all the assessed taste receptor genes (TAS1R1, TAS1R2, TAS1R3, SCNN1A and CD36) and taste tissue marker genes (NCAM1, GNAT3 and PLCβ2). Furthermore, quantitative expression was observed in a subset of taste genes assessed from the saliva collection techniques (cheek saliva, tongue saliva and Oragene RNA kit). These saliva collection techniques may be useful as a non-invasive alternative sampling technique to the fungiform papillae biopsy. Identification of the fungiform papillae biopsy as the optimal collection method will facilitate further research into understanding the effect of gene expression on variability in human taste perception.

Highlights

The presence of taste cells in the oral cavity is an essential component in our ability to taste the foods we consume
The taste cells express specialised receptors that sense the nutrient profile of ingested foods, with each taste cell tuned to a single taste quality i.e. sweet only, bitter only [1]
The papillae biopsy produced the highest RNA integrity (Table 2), with a RNA Integrity Number (RIN) average greater than 8, with the other collection techniques resulting in lower quality RNA

Summary

Introduction

The presence of taste cells in the oral cavity is an essential component in our ability to taste the foods we consume. The taste cells express specialised receptors that sense the nutrient profile of ingested foods, with each taste cell tuned to a single taste quality i.e. sweet only, bitter only [1]. Binding of a taste ligand to its cognate receptor (or ion through a taste associated ion channel) elicits a secondary messenger cascade within taste cells signalling the taste quality of the ingested food (i.e. sweet or bitter, respectively). While we detect the sweetness of a lollypop and the bitterness of tonic water, the perceived intensity varies significantly from person to person.

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