Implications of Notch Signaling in Taste Cell Replacement

Cyclophosphamide (CYP) was one of the first chemotherapy drugs developed and used to treat several types of cancer, by disrupting proliferative cells. Unfortunately, CYP is unable to differentiate between cancerous cells and healthy cells turning over which ultimately kills normally functioning cells, including those of the taste system. This loss of taste cells may result in dysgeusia (altered sense of taste), hypogeusia (reduced taste ability) or ageusia (inability to detect any tastes), eventually leading to malnutrition and poor prognosis for patients. The notch signaling pathway is one of the most important pathways involved in the differentiation and fate of neural stem cells (Hitoshi et al., 2002). A previous study looked at genes expressed in developing circumvallate taste cells and found that notch signaling remains active in adult mice to determine cell lineage as the sensory cells are continuously replaced (Seta, Seta, & Barlow, 2003). The current research uses immunohistochemistry to identify the presence of notch signaling following injury by CYP. It was hypothesized that if Notch1 is involved in taste cell replacement, we predict the Notch1 signal should be amplified following challenge by cyclophosphamide

The effect of cyclophosphamide on salt taste in mice

Chemotherapy is a common cancer treatment, yet it has many severe side effects including altered taste. Patients report that salt taste is most affected by chemotherapy. The salt taste transduction system has yet to be fully elucidated. Type I taste cells are thought to be responsible in part for salt taste. The goal of this study was to determine how cyclophosphamide (CYP), a common chemotherapeutic agent, affects salt taste in mice. This involved two experiments. The first experiment examined how an induced conditioned taste aversion (CTA) to NaCl (salt) would change following CYP treatment. The second used a brief access test to observe how NaCl preference changed before and after either a single dose or multiple dose CYP treatment. We hypothesized that CYP would affect Type I taste cells leading to changes in salt preference, that CYP would reduce salt aversion, and that multiple doses would affect multiple salt taste cell types leading to more significant changes in salt preference. Our results demonstrated that after treatment, CYP mice had higher NaCl lick rates than control mice. This occurred in two phases, initially around day 8 and again around day 18. CTA mice maintained an aversion to NaCl following treatment, indicating a pathway protected from CYP disturbance. A single CYP injection and multiple CYP injections had the same effects on mice, indicating that this methodology is not useful in disturbing multiple salt taste cell populations. These data support that there are at least two salt taste transduction pathways in mice

AP1 transcription factors are required to maintain 1 the peripheral taste system

The sense of taste is used by organisms to achieve the optimal nutritional requirement and avoid potentially toxic compounds. In the oral cavity, taste receptor cells are grouped together in taste buds that are present in specialized taste papillae in the tongue. Taste receptor cells are the cells that detect chemicals in potential food items and transmit that information to gustatory nerves that convey the taste information to the brain. As taste cells are in contact with the external environment, they can be damaged and are routinely replaced throughout an organism's lifetime to maintain functionality. However, this taste cell turnover loses efficiency over time resulting in a reduction in taste ability. Currently, very little is known about the mechanisms that regulate the renewal and maintenance of taste cells. We therefore performed RNA-sequencing analysis on isolated taste cells from 2 and 6-month-old mice to determine how alterations in the taste cell-transcriptome regulate taste cell maintenance and function in adults. We found that the activator protein-1 (AP1) transcription factors (c-Fos, Fosb and c-Jun) and genes associated with this pathway were significantly downregulated in taste cells by 6 months and further declined at 12 months. We generated conditional c-Fos-knockout mice to target K14-expressing cells, including differentiating taste cells. c-Fos deletion caused a severe perturbation in taste bud structure and resulted in a significant reduction in the taste bud size. c-Fos deletion also affected taste cell turnover as evident by a decrease in proliferative marker, and upregulation of the apoptotic marker cleaved-PARP. Thus, AP1 factors are important regulators of adult taste cell renewal and their downregulation negatively impacts taste maintenance

Dopaminergic and Non-Dopaminergic Value Systems in Conditioning and Outcome-Specific Revaluation

Animals are motivated to choose environmental options that can best satisfy current needs. To explain such choices, this paper introduces the MOTIVATOR (Matching Objects To Internal Values Triggers Option Revaluations) neural model. MOTIVATOR describes cognitiveemotional interactions between higher-order sensory cortices and an evaluative neuraxis composed of the hypothalamus, amygdala, and orbitofrontal cortex. Given a conditioned stimulus (CS), the model amygdala and lateral hypothalamus interact to calculate the expected current value of the subjective outcome that the CS predicts, constrained by the current state of deprivation or satiation. The amygdala relays the expected value information to orbitofrontal cells that receive inputs from anterior inferotemporal cells, and medial orbitofrontal cells that receive inputs from rhinal cortex. The activations of these orbitofrontal cells code the subjective values of objects. These values guide behavioral choices. The model basal ganglia detect errors in CS-specific predictions of the value and timing of rewards. Excitatory inputs from the pedunculopontine nucleus interact with timed inhibitory inputs from model striosomes in the ventral striatum to regulate dopamine burst and dip responses from cells in the substantia nigra pars compacta and ventral tegmental area. Learning in cortical and striatal regions is strongly modulated by dopamine. The model is used to address tasks that examine food-specific satiety, Pavlovian conditioning, reinforcer devaluation, and simultaneous visual discrimination. Model simulations successfully reproduce discharge dynamics of known cell types, including signals that predict saccadic reaction times and CS-dependent changes in systolic blood pressure.Defense Advanced Research Projects Agency and the Office of Naval Research (N00014-95-1-0409); National Institutes of Health (R29-DC02952, R01-DC007683); National Science Foundation (IIS-97-20333, SBE-0354378); Office of Naval Research (N00014-01-1-0624

Is fat the sixth taste primary? Evidence and implications

Explores our tongue\u27s ability to detect fat as a distinct taste similar to our ability to sense sweet, sour, bitter, acid and savory. Abstract Taste is the chemical sense responsible for the detection of non-volatile chemicals in potential foods. For fat to be considered as one of the taste primaries in humans, certain criteria must be met including class of affective stimuli, receptors specific for the class of stimuli on taste bud cells (TBC), afferent fibres from TBC to taste-processing regions of the brain, perception independent of other taste qualities and downstream physiological effects. The breakdown products of the macronutrients carbohydrates (sugars) and proteins (amino acids) are responsible for the activation of sweet and umami tastes, respectively. Following the same logic, the breakdown products of fat being fatty acids are the likely class of stimuli for fat taste. Indeed, psychophysical studies have confirmed that fatty acids of varying chain length and saturation are orally detectable by humans. The most likely fatty acid receptor candidates located on TBC are CD36 and G protein-coupled receptor 120. Once the receptors are activated by fatty acids, a series of transduction events occurs causing the release of neurotransmitters towards afferent fibres signalling the brain. Whether fatty acids elicit any direct perception independent of other taste qualities is still open to debate with only poorly defined perceptions for fatty acids reported. Others suggest that the fatty acid taste component is at detection threshold only and any perceptions are associated with either aroma or chemesthesis. It has also been established that oral exposure to fat via sham feeding stimulates increases in blood TAG concentrations in humans. Therefore, overall, with the exception of an independent perception, there is consistent emerging evidence that fat is the sixth taste primary. The implications of fatty acid taste go further into health and obesity research, with the gustatory detection of fats and their contributions to energy and fat intake receiving increasing attention. There appears to be a coordinated bodily response to fatty acids throughout the alimentary canal; those who are insensitive orally are also insensitive in the gastrointestinal tract and overconsume fatty food and energy. The likely mechanism linking fatty acid taste insensitivity with overweight and obesity is development of satiety after consumption of fatty foods

Qualitative and quantitative differences between taste buds of the rat and mouse

Abstract Background Numerous electrophysiological, ultrastructural, and immunocytochemical studies on rodent taste buds have been carried out on rat taste buds. In recent years, however, the mouse has become the species of choice for molecular and other studies on sensory transduction in taste buds. Do rat and mouse taste buds have the same cell types, sensory transduction markers and synaptic proteins? In the present study we have used antisera directed against PLCβ2, α-gustducin, serotonin (5-HT), PGP 9.5 and synaptobrevin-2 to determine the percentages of taste cells expressing these markers in taste buds in both rodent species. We also determined the numbers of taste cells in the taste buds as well as taste bud volume. Results There are significant differences (p 3) is smaller than a rat taste bud (64,200 μm3). The numerical density of taste cells in mouse circumvallate taste buds (2.1 cells/1000 μm3) is significantly higher than that in the rat (1.2 cells/1000 μm3). Conclusion These results suggest that rats and mice differ significantly in the percentages of taste cells expressing signaling molecules. We speculate that these observed dissimilarities may reflect differences in their gustatory processing.</p

Nicotine-Induced Effects on Nicotinic Acetylcholine Receptors (nAChRs), Ca2+ and Brain-Derived Neurotrophic Factor (BDNF) in STC-1 Cells

In addition to the T2R bitter taste receptors, neuronal nicotinic acetylcholine receptors (nAChRs) have recently been shown to be involved in the bitter taste transduction of nicotine, acetylcholine and ethanol. However, at present it is not clear if nAChRs are expressed in enteroendocrine cells other than beta cells of the pancreas and enterochromaffin cells, and if they play a role in the synthesis and release of neurohumoral peptides. Accordingly, we investigated the expression and functional role of nAChRs in enteroendocrine STC-1 cells. Our studies using RT-PCR, qRT-PCR, immunohistochemical and Western blotting techniques demonstrate that STC-1 cells express several α and β nAChR subunits. Exposing STC-1 cells to nicotine acutely (24h) or chronically (4 days) induced a differential increase in the expression of nAChR subunit mRNA and protein in a dose- and time-dependent fashion. Mecamylamine, a non-selective antagonist of nAChRs, inhibited the nicotineinduced increase in mRNA expression of nAChRs. Exposing STC-1 cells to nicotine increased intracellular Ca2+ in a dose-dependent manner that was inhibited in the presence of mecamylamine or dihydro-β-erythroidine, a α4β2 nAChR antagonist. Brain-derived neurotrophic factor (BDNF) mRNA and protein were detected in STC-1 cells using RT-PCR, specific BDNF antibody, and enzyme-linked immunosorbent assay. Acute nicotine exposure (30 min) decreased the cellular content of BDNF in STC-1 cells. The nicotine-induced decrease in BDNF was inhibited in the presence of mecamylamine. We also detected α3 and β4 mRNA in intestinal mucosal cells and α3 protein expression in intestinal enteroendocrine cells. We conclude that STC-1 cells and intestinal enteroendocrine cells express nAChRs. In STC-1 cells nAChR expression is modulated by exposure to nicotine in a doseand time-dependent manner. Nicotine interacts with nAChRs and inhibits BDNF expression in STC-1 cells

Biological activity of Citrus spp. metabolites on Ceratitis capitata (Wiedemann).

The Mediterranean fruit fly (medfly), Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), is one of the most injurious pest at global level. During the last years, several electrophysiological and behavioural studies have been carried out in order to investigate plant volatile compound-insect interactions with the aim to use this knowledge in sustainable control techniques. It has been observed that lemons are not attacked by medfly, probably because of the peel oil, that is toxic to other fruit flies. In the present paper electrophysiological recordings were conducted to evaluate the insect sensitivity to peel extract and peel oil of two Sicilian cultivars (Interdonato and Lunario) of Citrus x limon (L.) Burm.f. on C. capitata females. Behavioural bioassays were also performed to show their possible biological activity (repellent, antioviposition, insecticidal). C. limon peel extracts in different solvents (petroleum ether, dichloromethane and methanol) were investigated at various concentrations using a single cell recording technique (stimulation of tarsal taste chemosensilla). Different tarsal taste cell responses to the two cultivars were recorded. The higher sensitivity was evoked by C. limon Interdonato, particularly to the methanol extract, which elicited significant increases in the spike frequency at increasing concentrations. The peel oil of the same cultivars as well as that ones of other two C. limon varieties (Monachello and Femminello) have been tested by EAG techniques. The EAG data showed a high sensitivity (about -8.0/8.5mV) of the medfly antennae to the oils of Citrus spp. and a clear dose-response relationship. Responses of adult females (virgin and mated) to Citrus spp. peel extract were quantified in a double-choice test using yellow spheres (diameter 7.0cm) housed in field cages. Preliminary tests conducted on three extracts of C. limon Interdonato and Lunario have provided interesting results. It was recorded a general decrease of the oviposition on treated spheres compared to control and in the case of the cultivar Lunario, a mortality of insects

The Evolution of Senses: My Research Journey into the Nervous System of Cnidaria

Our understanding of the evolutionary history of animals is improving, but knowledge of the ancient sensory systems that early animals used to interact with their environments is still largely unknown. Using molecular cloning and in situ hybridization staining procedures, I was able to test the hypothesis that some senses evolved prior to the evolution of animals with bilateral symmetry. My data provides evidence that cnidarians can taste using genes that are closely related to human taste receptors. This finding changes our current understanding of when tasteevolved by hundreds of millions of years. The in situ hybridization results also demonstrated co-localization, or overlap, of the expression of taste and photosensitivity genes, which provides preliminary evidence that cnidarians use a polymodal sensory-motor (PSM) neuron to sense light and chemical cues (“tastes”) to coordinate their feeding behavior. The cDNA constructs I have produced will also provide further biochemical insights into their function. My long-term research projects have taught me about the process of making scientific discoveries, and I hope to continue conducting research throughout my career