Cells in taste buds are closely packed with little extracellular space. Tight junctions and other barriers further limit permeability and may result in buildup of extracellular K+ following action potentials. In many tissues, inwardly-rectifying K channels such as the Renal Outer Medullary K (ROMK) channel (also called Kir1.1 and derived from the Kcnj1 gene) help to redistribute K+. Using RT-PCR, we defined ROMK splice variants in mouse kidney, and report here the expression of a single one of these, ROMK2, in a subset of mouse taste cells. With qRT-PCR, we show the abundance of ROMK mRNA in taste buds is vallate > foliate ≫ palate ≫ fungiform. ROMK protein follows the same pattern of prevalence as mRNA, and is essentially undetectable by immunohistochemistry in fungiform taste buds. ROMK protein is localized to the apical tips of a subset of taste cells. Using tissues from PLCβ2-GFP and GAD-GFP transgenic mice, we show that ROMK is not found in PLCβ2-expressing type II/Receptor cells or in GAD-expressing type III/Presynaptic cells. Instead, ROMK is found, by single-cell RT-PCR and immunofluorescence, in most cells that are positive for the taste glial cell marker, Ectonucleotidase2. ROMK is precisely localized to the apical tips of these cells, at and above apical tight junctions. We propose that in taste buds, ROMK in type I/glial-like cells may serve a homeostatic function, excreting excess K+ through the apical pore, and allowing excitable taste cells to maintain a hyperpolarized resting membrane potential.

The lateral habenula (LHb) plays an important role in learning driven by negative outcomes. Many drugs of abuse, including ethanol, have dose-dependent aversive effects that act to limit intake of the drug. However, the role of the LHb in regulating ethanol intake is unknown. In the present study, we compared voluntary ethanol consumption and self-administration, yohimbine-induced reinstatement of ethanol seeking, and ethanol-induced conditioned taste aversion in rats with sham or LHb lesions. In rats given home cage access to 20% ethanol in an intermittent access two bottle choice paradigm, lesioned animals escalated their voluntary ethanol consumption more rapidly than sham-lesioned control animals and maintained higher stable rates of voluntary ethanol intake. Similarly, lesioned animals exhibited higher rates of responding for ethanol in operant self-administration sessions. In addition, LHb lesion blocked yohimbine-induced reinstatement of ethanol seeking after extinction. Finally, LHb lesion significantly attenuated an ethanol-induced conditioned taste aversion. Our results demonstrate an important role for the LHb in multiple facets of ethanol-directed behavior, and further suggest that the LHb may contribute to ethanol-directed behaviors by mediating learning driven by the aversive effects of the drug.

BackgroundThe neuropeptide, oxytocin (OXT), acts on brain circuits to inhibit food intake. Mutant mice lacking OXT (OXT knockout) overconsume salty and sweet (i.e. sucrose, saccharin) solutions. We asked if OXT might also act on taste buds via its receptor, OXTR.Methodology/Principal FindingsUsing RT-PCR, we detected the expression of OXTR in taste buds throughout the oral cavity, but not in adjacent non-taste lingual epithelium. By immunostaining tissues from OXTR-YFP knock-in mice, we found that OXTR is expressed in a subset of Glial-like (Type I) taste cells, and also in cells on the periphery of taste buds. Single-cell RT-PCR confirmed this cell-type assignment. Using Ca2+ imaging, we observed that physiologically appropriate concentrations of OXT evoked [Ca2+]i mobilization in a subset of taste cells (EC50 ∼33 nM). OXT-evoked responses were significantly inhibited by the OXTR antagonist, L-371,257. Isolated OXT-responsive taste cells were neither Receptor (Type II) nor Presynaptic (Type III) cells, consistent with our immunofluorescence observations. We also investigated the source of OXT peptide that may act on taste cells. Both RT-PCR and immunostaining suggest that the OXT peptide is not produced in taste buds or in their associated nerves. Finally, we also examined the morphology of taste buds from mice that lack OXTR. Taste buds and their constituent cell types appeared very similar in mice with two, one or no copies of the OXTR gene.Conclusions/SignificanceWe conclude that OXT elicits Ca2+ signals via OXTR in murine taste buds. OXT-responsive cells are most likely a subset of Glial-like (Type I) taste cells. OXT itself is not produced locally in taste tissue and is likely delivered through the circulation. Loss of OXTR does not grossly alter the morphology of any of the cell types contained in taste buds. Instead, we speculate that OXT-responsive Glial-like (Type I) taste bud cells modulate taste signaling and afferent sensory output. Such modulation would complement central pathways of appetite regulation that employ circulating homeostatic and satiety signals.

Oxytocin (OXT) suppresses food intake and lack of OXT leads to overconsumption of sucrose. Taste bud cells were recently discovered to express OXT-receptor. In the present study we tested whether administering OXT to wild-type mice affects their licking behavior for tastants in a paradigm designed to be sensitive to taste perception. We injected C57BL/6J mice intraperitoneally (i.p.) with 10 mg/kg OXT and assayed their brief-access lick responses, motivated by water deprivation, to NaCl (300 mM), citric acid (20 mM), quinine (0.3 mM), saccharin (10 mM), and a mix of MSG and IMP (100 mM and 0.5 mM respectively). OXT had no effect on licking for NaCl, citric acid, or quinine. A possible effect of OXT on saccharin and MSG+IMP was difficult to interpret due to unexpectedly low lick rates to water (the vehicle for all taste solutions), likely caused by the use of a high OXT dose that suppressed licking and other behaviors. A subsequent experiment focused on another preferred tastant, sucrose, and employed a much lower OXT dose (0.1 mg/kg). This modification, based on our measurements of plasma OXT following i.p. injection, permitted us to elevate plasma [OXT] sufficiently to preferentially activate taste bud cells. OXT at this low dose significantly reduced licking responses to 0.3 M sucrose, and overall shifted the sucrose concentration – behavioral response curves rightward (mean EC50saline = 0.362 M vs. EC50OXT = 0.466 M). Males did not differ from females under any condition in this study. We propose that circulating oxytocin is another factor that modulates taste-based behavior.