Genetic Labeling of Car4-expressing Cells Reveals Subpopulations of Type III Taste Cells

Chem Senses. 2017 Oct 31;42(9):747-758. doi: 10.1093/chemse/bjx048.

Abstract

Carbonic anhydrases form an enzyme family of 16 members, which reversibly catalyze the hydration of carbon dioxide to bicarbonate and protons. In lung, kidney, and brain, presence of carbonic anhydrases is associated with protons and bicarbonate transport in capillary endothelium of lung, reabsorption of bicarbonate in proximal renal tubules, and extracellular buffering. In contrast, their role in taste is less clear. Recently, carbonic anhydrase IV expression was detected in sour-sensing presynaptic taste cells and was associated with the taste of carbonation, yet the precise role and cell population remained uncertain. To examine the role of carbonic anhydrase 4-expressing cells in taste reception, we generated a mouse strain carrying a modified allele of the carbonic anhydrase 4 gene in which the coding region of the red fluorescent protein monomeric Cherry is attached to that of carbonic anhydrase 4 via an internal ribosome entry site. Monomeric Cherry fluorescence was detected in lingual papillae as well as taste buds of soft palate and naso-incisor duct. However, expression patterns on the tongue differ between posterior and fungiform papillae. Whereas monomeric Cherry auto-fluorescence was almost always co-localized with presynaptic cell markers aromatic L-amino-acid decarboxylase, synaptosomal-associated protein 25 or glutamic acid decarboxylase 67 in fungiform papillae and taste buds of palate and naso-incisor duct, monomeric Cherry-positive cells in posterior tongue papillae represent only a subpopulation of presynaptic cells. We conclude that this model is well suited for detailed investigation into the role of carbonic anhydrase in gustation and other processes.

Keywords: carbonic anhydrase 4; gene-targeted animals; knock-in; mCherry; presynaptic cell; sour taste.

MeSH terms

  • Animals
  • Calcium Channels / genetics
  • Calcium Channels / metabolism
  • Carbonic Anhydrases / genetics
  • Carbonic Anhydrases / metabolism*
  • Gene Knock-In Techniques
  • Genetic Engineering
  • In Situ Hybridization
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Mice
  • Mice, 129 Strain
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / metabolism
  • Red Fluorescent Protein
  • Taste Buds / cytology
  • Taste Buds / metabolism*
  • Tongue / metabolism
  • Tongue / pathology

Substances

  • Calcium Channels
  • Luminescent Proteins
  • Pkd2l1 protein, mouse
  • Receptors, Cell Surface
  • Carbonic Anhydrases
  • carbonic anhydrase VI