A5021571464- Biochemical Analysis and Sensing Techniques
- Olfactory and Sensory Function Studies
- Animal Genetics and Reproduction
- Advanced Chemical Sensor Technologies
- Virus-based gene therapy research
- CRISPR and Genetic Engineering
- Neuroscience and Neuropharmacology Research
- RNA and protein synthesis mechanisms
- Xenotransplantation and immune response
- Alzheimer's disease research and treatments
- Regulation of Appetite and Obesity
- Neurobiology and Insect Physiology Research
- Connexins and lens biology
- Toxin Mechanisms and Immunotoxins
- Photochromic and Fluorescence Chemistry
- Drug Transport and Resistance Mechanisms
- Ion Channels and Receptors
- Neuroinflammation and Neurodegeneration Mechanisms
- Neuroendocrine regulation and behavior
- Healthcare and Venom Research
- Genetically Modified Organisms Research
- Cholesterol and Lipid Metabolism
- Myasthenia Gravis and Thymoma
- Peripheral Neuropathies and Disorders
- Lipid Membrane Structure and Behavior
University of Miami
2009-2022
Zero to Three
2009
Institute of Molecular Biology and Biophysics
2007
Universidade Federal do Rio de Janeiro
2005
Mental Health Research Center of Russian Academy of Medical Sciences
2002
Louisiana State University
2002
Institute of Molecular Genetics
2002
Boston University
2002
Russian Academy of Sciences
2002
Federal Research Centre of Nutrition and Biotechnology
2001
ATP has been shown to be a taste bud afferent transmitter, but the cells responsible for, and mechanism of, its release have not identified. Using CHO expressing high-affinity neurotransmitter receptors as biosensors, we show that gustatory stimuli cause receptor secrete through pannexin 1 hemichannels in mouse buds. further stimulates other second serotonin. These results provide link intracellular Ca(2+) during transduction secretion of ATP, from cells. They also indicate route for...
Taste buds are aggregates of 50–100 cells, only a fraction which express genes for taste receptors and intracellular signaling proteins. We combined functional calcium imaging with single-cell molecular profiling to demonstrate the existence two distinct cell types in mouse buds. Calcium revealed that isolated cells responded transient elevation cytoplasmic Ca 2+ either tastants or depolarization KCl, but never both. Using reverse transcription (RT)-PCR, we show individual phospholipase C β2...
Abstract Gustatory stimuli are detected by taste buds and transmitted to the hindbrain via sensory afferent neurons. Whether each quality (sweet, bitter so on) is encoded separate neurons (‘labelled lines’) remains controversial. We used mice expressing GCaMP3 in geniculate ganglion investigate taste-evoked activity. Using confocal calcium imaging, we recorded responses oral stimulation with prototypic stimuli. Up 69% of respond multiple tastants. Moreover, tuned a single at low...
Mammalian taste buds use ATP as a neurotransmitter. Taste Receptor (type II) cells secrete via gap junction hemichannels into the narrow extracellular spaces within bud. This excites primary sensory afferent fibers and also stimulates neighboring bud cells. Here we show that is enzymatically degraded to adenosine mouse vallate this nucleoside acts an autocrine neuromodulator selectively enhance sweet taste. In in lingual slice preparation, Ca 2+ mobilization evoked by focally applied...
Taste buds are innervated by neurons whose cell bodies reside in cranial sensory ganglia. Studies on the functional properties and connectivity of these hindered lack markers to define their molecular identities classes. The mouse geniculate ganglion contains chemosensory innervating lingual palatal taste somatosensory pinna. Here, we report single RNA sequencing neurons. Using unbiased transcriptome analyses, show a pronounced separation between two major clusters which, anterograde...
Sour (acid) taste is postulated to result from intracellular acidification that modulates one or more acid-sensitive ion channels in receptor cells. The identity of such channel(s) remains uncertain. Potassium channels, by regulating the excitability cells, are candidates for acid transducers. Several 2-pore domain potassium leak conductance (K(2)P family) sensitive acidification. We examined their expression mouse vallate and foliate buds using RT-PCR, detected TWIK-1 -2, TREK-1 TASK-1. Of...
Abstract Background "Type II"/Receptor cells express G protein-coupled receptors (GPCRs) for sweet, umami (T1Rs and mGluRs) or bitter (T2Rs), as well the proteins downstream signalling cascades. Transduction of T1Rs T2Rs relies on G-protein PLCβ2-mediated release stored Ca 2+ . Whereas Gαgus (gustducin) couples to T2R (bitter) receptors, which Gα-subunit sweet (T1R2 + T1R3) receptor is presently not known. We utilized RT-PCR, immunocytochemistry single-cell gene expression profiling examine...
Abstract Although adenosine triphosphate (ATP) is known to be an afferent transmitter in the peripheral taste system, serotonin (5‐HT) and norepinephrine (NE) have also been proposed as candidate neurotransmitters detected immunocytochemically mammalian cells. To understand significance of biogenic amines taste, we evaluated ability cells synthesize, transport, package 5‐HT NE. We show by reverse transcriptase‐polymerase chain reaction immunofluorescence microscopy that enzymes for...
Abstract Cells in taste buds are closely packed, with little extracellular space. Tight junctions and other barriers further limit permeability may result buildup of K + following action potentials. In many tissues, inwardly rectifying channels such as the renal outer medullary (ROMK) channel (also called Kir1.1 derived from Kcnj1 gene) help to redistribute . Using reverse‐transcription polymerase chain reaction (RT‐PCR), we defined ROMK splice variants mouse kidney report here expression a...
Taste buds consist of at least three principal cell types that have different functions in processing gustatory signals: glial-like (type I) cells, receptor II) and presynaptic III) cells. Using a combination Ca 2+ imaging, single-cell reverse transcriptase-PCR immunostaining, we show GABA is an inhibitory transmitter mouse taste buds, acting on A B receptors to suppress (ATP) secretion from cells during stimulation. Specifically, express subunits β2, δ, π, as well receptors. In contrast,...
The acid-sensitive cation channel acid-sensing ion channel-2 (ASIC2) is widely believed to be a receptor for acid (sour) taste in mammals on the basis of its physiological properties and expression rat bud cells. Using reverse transcriptase-PCR, we detected ASIC1 ASIC3, but not ASIC4, mouse buds nonsensory lingual epithelium. Surprisingly, did detect mRNA ASIC2 buds, although readily observed buds. Furthermore, Ca 2+ imaging experiments, knock-out mice exhibited normal responses (increases...
In response to gustatory stimulation, taste bud cells release a transmitter, ATP, that activates P2X2 and P2X3 receptors on afferent fibers. Taste behavior neural responses are largely abolished in mice lacking [P2X2 double knock-out (DKO) mice]. The assumption has been eliminating only removes postsynaptic targets but transmitter secretion is normal. Using functional imaging, ATP biosensor cells, cell-free assay for we tested this assumption. Surprisingly, although stimulation mobilizes Ca...
Background The 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 might also act taste buds via its receptor, OXTR. Methodology/Principal Findings Using RT-PCR, we detected the expression of OXTR in throughout oral cavity, but not adjacent non-taste lingual epithelium. By immunostaining tissues from OXTR-YFP knock-in mice, found that is expressed a...
In mammalian taste buds, Type I cells comprise half of all cells. These are termed "glial-like" based on morphologic and molecular features, but there limited studies describing their function. We tested whether sense chemosensory activation adjacent (i.e., Types II III) bud cells, similar to synaptic glia. Using Gad2;;GCaMP3 mice both sexes, we confirmed by immunostaining that, within GCaMP expression is predominantly in (with no ≈28% III expressing weakly). dissociated GCaMP+ responded...
The second messenger, 3',5'-cyclic adenosine monophosphate (cAMP), is known to be modulated in taste buds following exposure gustatory and other stimuli. Which cell type(s) (Type I/glial-like cells, Type II/receptor or III/presynaptic cells) undergo taste-evoked changes of cAMP what the functional consequences such are remain unknown. Using Fura-2 imaging isolated mouse vallate we explored how elevating alters Ca(2+) levels identified cells. Stimulating with forskolin (Fsk; 1 microm) +...
The production of recombinant human granulocyte-colony stimulating factor (hG-CSF) for therapeutic purposes relies on its expression in selected clones transfected mammalian cells. Alternatively, this protein can be produced by targeted secretion the milk transgenic goats. Thus, aim study was to produce founder goats expressing hG-CSF and propagate a herd. After DNA microinjection vivo-produced pronuclear embryos performed, two founders were obtained (one male one female, named 10 M 12 F,...
Abstract Because of their ease use, adeno-associated viruses (AAVs) are indispensable tools for much neuroscience. Yet AAVs have been used relatively little to study the identities and connectivity peripheral sensory neurons, principally because methods selectively target neurons limited. The introduction AAV-PHP.S capsid with enhanced tropism (Chan et al., 2017) offered a solution, which we further elaborate here. Using GFP or mScarlet fluorescent proteins, show that mouse ganglia cranial...
ABSTRACT Because of their ease use and low risk containment, Adeno-Associated Virus vectors are indispensable tools for much neuroscience. Yet AAVs have been used relatively little to study the identities connectivity peripheral sensory neurons because methods selectively target particular receptive fields or neuron types limited. The introduction AAV-PHP.S capsid with selective tropism (Chan et al., 2017) offered a solution, which we further elaborate here. We demonstrate using GFP mScarlet...