Biomedical subjects

Roberto Tirindelli

Publications and source records attributed to Roberto Tirindelli.

7 recordsLinked to original sources

PubMed2006

Neuropeptide Y in the olfactory microvillar cells.

This paper examines a possible role of microvillar cells in coordinating cell death and regeneration of olfactory epithelial neurons. The olfactory neuroepithelium of mammals is a highly dynamic organ. Olfactory neurons periodically degenerate by apoptosis and as a consequence of chemical or physical damage. To compensate for this loss of cells, the olfactory epithelium maintains a lifelong ability to regenerate from a pool of resident multipotent stem cells. To assure functional continuity and histological integrity of the olfactory epithelium over a period of many decades, apoptosis and regeneration require to be precisely coordinated. Among the factors that have been implicated in mediating this regulation is the neuropeptide Y (NPY). Knockout mice that lack functional expression of this neurogenic peptide show defects in embryonic development of the olfactory epithelium and in its ability to regenerate in the adult. Here we show that, in postnatal olfactory epithelia, NPY is exclusively expressed by a specific population of microvillar cells. We previously characterized these cells as a novel type of putative chemosensory cells, which are provided with a phosphatidyl-inositol-mediated signal transduction cascade. Our findings allow for the first time to suggest that microvillar cells are involved in connecting apoptosis to neuronal regeneration by stimulus-induced release of NPY.

Animals↗

PubMed2006

Cross-modal interactions between olfaction and vision when grasping.

This study used kinematics to investigate the integration between vision and olfaction during grasping movements. Participants were requested to smell an odorant and then grasp an object presented in central vision. The results indicate that if the target was small (e.g., a strawberry), the time and amplitude of maximum hand aperture were later and greater, respectively, when the odor evoked a larger object (e.g., an orange) than when the odor evoked an object of a similar size as the target or no odor was presented. Conversely, the time and amplitude of maximum hand aperture were earlier and reduced, respectively, when the target was large (e.g., a peach) and the odor evoked a smaller sized object (e.g., an almond) than when the odor evoked an object of a similar size as the target or no odor was presented. Taken together, these results support the evidence of cross-modal links between olfaction and vision and extend this notion to goal-directed actions.

Adult↗

PubMed2005

The vomeronasal receptor V2R2 does not require escort molecules for expression in heterologous systems.

In rodents, many behavioural responses are triggered by pheromones. These molecules are believed to bind and activate two families of G-protein coupled receptors, namely V1Rs and V2Rs, which are specifically expressed in the chemosensory neurons of the vomeronasal organ. V2Rs are homologous with Group 3 of G-protein-coupled receptors, which includes metabotropic glutamate receptors, calcium-sensing receptors, fish olfactory receptors, and taste receptors for sweet molecules and amino acids. The large extracellular region of these receptors is folded as a dimer and, in this form, binds agonists that in many cases are amino acids. It has recently been reported that V2Rs must be physically associated with specific major histocompatibility complex class Ib molecules (MHC) for their expression in both mouse vomeronasal neurons and heterologous cell lines. Here, we show that in contrast to the other V2Rs, V2R2, an atypical member of this receptor family, can be successfully and abundantly expressed by insect cells without the requirement of escort molecules like MHC. Moreover, the extracellular binding domain of V2R2, secreted as a soluble product, forms dimers via cysteine-mediated sulphur bridges. Overall, the data presented in this paper confirm that V2R2 diverges from the other members of the V2R family and suggest a different role for this receptor in pheromonal communication.

Animals↗

PubMed2005

Phosphatidyl-inositide signalling proteins in a novel class of sensory cells in the mammalian olfactory epithelium.

Ciliated sensory neurons, supporting cells and basal stem cells represent major cellular components of the main olfactory epithelium in mammals. Here we describe a novel class of sensory cells in the olfactory neuroepithelium. The cells express phospholipase C beta-2 (PLC beta2), transient receptor potential channels 6 (TRPC6) and inositol 3, 4, 5-trisphosphate receptors type III (InsP3R-III). Unlike ciliated olfactory neurons, they express neither olfactory marker protein nor centrin, adenylyl cyclase or cyclic nucleotide-gated cation channels. Typical components of the cytoskeleton of microvilli, ezrin and actin are found co-localized with PLC beta2 and TRPC6 in apical protrusions of the cells. In Ca2+-imaging experiments, the cells responded to odours. They express neuronal marker proteins and possess an axon-like process, but following bulbectomy the cells do not degenerate. Our results suggest a novel class of microvillous secondary chemosensory cells in the mammalian olfactory system. These cells, which utilize phosphatidyl-inositides in signal transduction, represent about 5% of all olfactory cells. Their abundance indicates that they play an important role in stimulus-dependent functions and/or the regeneration of the olfactory system.

Animals↗

PubMed2003

Apical and basal neurones isolated from the mouse vomeronasal organ differ for voltage-dependent currents.

The mammalian vomeronasal organ (VNO) contains specialized neurones that transduce the chemical information related to pheromones into discharge of action potentials to the brain. Molecular and biochemical studies have shown that specific components of the pheromonal transduction systems are segregated into two distinct subsets of vomeronasal neurones: apical neurones and basal neurones. However, it is still unknown whether these neuronal subsets also differ in other functional characteristics, such as their membrane properties. We addressed this issue by studying the electrophysiological properties of vomeronasal neurones isolated from mouse VNO. We used the patch-clamp technique to examine both the passive membrane properties and the voltage-gated Na+, K+ and Ca2+ currents. Apical neurones were distinguished from basal ones by the length of their dendrites and by their distinct immunoreactivity for the putative pheromone receptor V2R2. The analysis of passive properties revealed that there were no significant differences between the two neuronal subsets. Also, apical neurones were similar to basal neurones in their biophysical and pharmacological properties of voltage-gated Na+ and K+ currents. However, we found that the density of Na+ currents was about 2-3 times greater in apical neurones than in basal neurones. Consistently, in situ hybridization analysis revealed a higher expression of the Na+ channel subtype III in apical neurones than in basal ones. In contrast, basal neurones were endowed with Ca2+ currents (T-type) of greater magnitude than apical neurones. Our findings indicate that apical and basal neurones in the VNO exhibit distinct electrical properties. This might have a profound effect on the sensory processes occurring in the VNO during pheromone detection.

Action Potentials↗

PubMed2003

Ion conductances in supporting cells isolated from the mouse vomeronasal organ.

The vomeronasal organ (VNO) is a chemosensory structure involved in the detection of pheromones in most mammals. The VNO sensory epithelium contains both neurons and supporting cells. Data suggest that vomeronasal neurons represent the pheromonal transduction sites, whereas scarce information is available on the functional properties of supporting cells. To begin to understand their role in VNO physiology, we have characterized with patch-clamp recording techniques the electrophysiological properties of supporting cells isolated from the neuroepithelium of the mouse VNO. Supporting cells were distinguished from neurons by their typical morphology and by the lack of immunoreactivity for Ggamma8 and OMP, two specific markers for vomeronasal neurons. Unlike glial cells in other tissues, VNO supporting cells exhibited a depolarized resting potential (about -29 mV). A Goldman-Hodgkin-Katz analysis for resting ion permeabilities revealed indeed an unique ratio of P(K):P(Na):P(Cl) = 1:0.23:1.4. Supporting cells also possessed voltage-dependent K(+) and Na(+) conductances that differed significantly in their biophysical and pharmacological properties from those expressed by VNO neurons. Thus glial membranes in the VNO can sustain significant fluxes of K(+) and Na(+), as well as Cl(-). This functional property might allow supporting cells to mop-up and redistribute the excess of KCl and NaCl that often occurs in certain pheromone-delivering fluids, like urine, and that could blunt the sensitivity of VNO neurons to pheromones. Therefore vomeronasal supporting cells could affect chemosensory transduction in the VNO by regulating the ionic strength of the pheromone-containing medium.

Animals↗

PubMed1991

Characterization and Regional Distribution of a Class of Synapses with Highly Concentrated cAMP Binding Sites in the Rat Brain.

A class of putative synaptic terminals with concentrated cAMP binding sites are labelled in unfixed sections of rat brain by means of the ligand 8-thioacetamido fluorescein cAMP (SAF-cAMP), a fluorescent analogue of cAMP. The labelled terminals appear as sharply delimited bouton-like structures in close proximity but external to the cell body of neurons. The SAF-cAMP binding, measured at equilibrium in competition with other nucleotides, indicates that the binding site recognizes the cAMP moiety of SAF-cAMP. In the labelled terminals of the frontal cortex the concentration of SAF-cAMP binding sites is estimated to be in the millimolar range (at least 2.1 +/- 1.0 mM). In a brain homogenate, labelled terminals are visualized only in the membrane fraction enriched in synaptosomes. The cAMP binding activity of the synaptosomes is insoluble in high and in low ionic strength solution and is only partially solubilized by detergents, suggesting that the binding sites are intrinsic membrane proteins and/or proteins associated with the cytoskeleton. There is the possibility that SAF-cAMP labels new cAMP binding sites highly concentrated in a class of synaptic terminals. SAF-cAMP labelling is prominent in well defined regions of the rat brain: (i) the frontal and entorhinal areas of the cortex; (ii) the field CA1 of the hippocampus; (iii) the olfactory system; (iv) the medial nuclei of the thalamus; (v) the parabrachial nuclei and other less defined regions of the reticular substance; (vi) the substantia gelatinosa of Rolando in the spinal cord; and (vii) the neo- and paleocerebellum in the Purkinje cell layer, the archicerebellum in the granular cell layer. SAF-cAMP labelling is absent in specific motor and sensory structures, with the exception of the olfactory system. It is proposed that SAF-cAMP binding sites single out a new type of synaptic terminals involved in complex nervous functions.

Journal Article↗