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S Firestein

Publications and source records attributed to S Firestein.

At least 37 records · Page 2Linked to original sources
PubMed1995

Quantal-like current fluctuations induced by odorants in olfactory receptor cells.

Many sensory systems have evolved signal detection capabilities that are limited only by the physical attributes of the stimulus. For example, 'hair' cells of the inner ear can detect displacements of atomic dimensions. Likewise, both in vertebrates and in invertebrates photoreceptors can detect a single photon. The olfactory stimulus also has a quantal unit, the single odorant molecule. Insects are reportedly able to detect a single pheromone molecule, whereas quantal responses in vertebrate olfactory receptor cells have not been reported yet. Psychophysical measurements indicate that a minimum of 50 odorant molecules are necessary for human olfactory detection, suggesting that an individual receptor may be activated by a single odorant molecule. We report here measurements of current fluctuations induced by odorants that suggest a quantal event of about 0.3-1 pA, presumably triggered by the binding of a single odorant molecule.

Ambystoma↗
PubMed1995

Interaction of anionic and cationic currents leads to a voltage dependence in the odor response of olfactory receptor neurons.

1. We recorded odor-induced currents from isolated olfactory receptor neurons of the land phase tiger salamander (Ambystoma tigrinum) with the whole cell patch clamp. 2. In a subset of cells the current-voltage relation for the odor-induced current showed a strong rectification with, in some cells, a negative resistance slope between about -45 and -25 mV. In these cells there was little or no odor-induced current at -55 mV, the average resting potential of olfactory neurons. 3. Depolarizing the membrane to +20 mV revealed a large outward current, and on repolarizing the membrane to -55 mV we could observe a large inward current. This current was not observed in the absence of the depolarizing step or in the absence of odor stimuli. 4. This odor-induced tail current was dependent on extracellular Ca2+ and voltage, activating with increased depolarization. The reversal potential was sensitive to the chloride equilibrium potential and it could be significantly blocked by niflumic acid, a blocker of calcium-activated chloride currents. The voltage dependence could result from either the voltage-dependent block of adenosine 3',5'-cyclic monophosphate-gated cation channels known to be activated by odorants and permeable to Ca2+, or from an inherent voltage dependence in the chloride channel gating. 5. The current appears to function as a regenerative mechanism that might increase the amplitude and duration of the odor-induced current, especially to low concentrations of stimulus.

Ambystoma↗
PubMed1994

NADPH-diaphorase localization in the olfactory system.

NADPH-diaphorase staining was studied in the intact olfactory system and following transections of the olfactory nerve. Intense staining was found in the olfactory epithelia of rat and salamander, especially at the mucosal surface. The olfactory bulb nerve and glomerular layers were also stained. Transection of the olfactory nerve in salamander, or bulbectomy in rat, resulted in decreased staining at the mucosal surface, although staining increased in cells deeper in the epithelium. Deafferentation also caused the disappearance of staining in the olfactory nerve and glomerular layers of the bulb. These results support the notion of a role for NADPH in the olfactory system, perhaps as a biosynthesis substrate for NO modulation of cGMP in the developing epithelium and a complementary role for CO in the adult.

Animals↗
PubMed1994

The cyclic nucleotide gated channel of olfactory receptor neurons.

Olfactory transduction proceeds through a G-protein coupled cascade that produces the ubiquitous second messenger cyclic AMP. The cyclic AMP causes a change in membrane potential by acting directly on an ion channel that allows cations to flow into the cell. This ion channel is one of a new family of ion channels that are activated by intracellular cyclic nucleotides. However, even though they are activated by binding a ligand their amino acid structure shows that they share a common ancestry with voltage activated channels, especially voltage dependent Ca2+ channels. In olfactory neurons these channels perform a critical role in the transduction of chemical information in the environment into changes in membrane electrical properties that are transmitted to higher order processing centers in the brain.

Animals↗
PubMed1993

Rapid application and removal of second messengers to cyclic nucleotide-gated channels from olfactory epithelium.

The last step in the second-messenger cascade mediating vertebrate olfactory transduction is the direct opening of a nonspecific cation channel by cAMP. The kinetic properties of this interaction are critical in determining the time course of the sensory response. To analyze these properties, excised inside-out membrane patches containing either the native channel from salamander olfactory-receptor neurons or a recombinant rat olfactory cyclic nucleotide-gated channel were exposed to short pulses of known concentrations of cAMP or cGMP to mimic a rapid and transient production of second messenger. Channel activity outlasted cyclic nucleotide pulses for several hundred milliseconds. This effect was due to an intrinsic property of the olfactory channel protein because it did not occur with cGMP-activated channels from retinal photoreceptors. Gating kinetics of the olfactory channel were both voltage and agonist dependent. These results demonstrate that the overall slow channel-gating kinetics could account for the difference in time course between the odor-induced changes in cAMP concentration and the subsequent sensory generator current.

Ambystoma↗
PubMed1993

Membrane currents and mechanisms of olfactory transduction.

The term olfactory transduction refers to the mechanisms that transform chemical information into electrical signals. With the patch-clamp technique it is possible to record those signals and to infer something about the mechanism that produced them. The direct activation of a cation-permeable channel by cAMP is the final step in producing the odour-induced ionic current. Because it occupies a critical position in the transduction process, measurements of the ion channel's activity provide useful insights into the molecular processes underlying olfactory transduction. In addition to its activation by cAMP and cGMP, the channel is modulated by both extracellular and intracellular Ca2+ ions and by extracellular Mg2+ ions, all at physiological concentrations. These effects are probably important in promoting signal reliability. An unusual feature of this channel is its termination kinetics--it can remain active for hundreds of milliseconds after the agonist has been removed. This is likely to add to the integrating properties of the olfactory sensory neuron.

Animals↗
PubMed1993

The relation between stimulus and response in olfactory receptor cells of the tiger salamander.

1. Olfactory receptor cells were isolated from the adult tiger salamander Ambystoma tigrinum and the current in response to odorant stimuli was measured with the whole-cell voltage-clamp technique while odorants at known concentrations were rapidly applied for controlled exposure times. 2. Three odorants, cineole, isoamyl acetate and acetophenone, were first applied at 5 x 10(-4) M. Out of forty-nine cells tested, 53% responded to one odorant only, 22% to two odorants and 25% to all three odorants. 3. The amplitude of the current in response to a given odorant concentration was found to be dependent on the duration of the odorant stimulus and reached a saturating peak value at 1.2 s of stimulus duration. 4. The current measured at the peak of the response for odorant steps of 1.2 s as a function of odorant concentration was well described by the Hill equation for the three odorants with Hill coefficients higher than 1 and K1/2 (odorant concentration needed to activate half the maximal current) ranging from 3 x 10(-6) to 9 x 10(-5) M. 5. It is concluded that olfactory receptor cells are broadly tuned and have a low apparent affinity for odorants, integrate stimulus information over time, and have a narrow dynamic range.

Acetophenones↗
PubMed1993

Divalent cations block the cyclic nucleotide-gated channel of olfactory receptor neurons.

1. The effects of external divalent cations on odor-dependent, cyclic AMP-activated single-channel currents from olfactory receptor neurons of the tiger salamander (Ambystoma tigrinum) were studied in inside-out membrane patches taken from dendritic regions of freshly isolated sensory cells. 2. Channels were reversibly activated by 100 microM cyclic AMP. In the absence of divalent cations, the channel had a linear current-voltage relation giving a conductance of 45 pS. With increasing concentrations of either Ca2+ or Mg2+ in the external solution, the channel displayed a rapid flickering behavior. At higher concentrations of divalent cations, the transitions were too rapid to be fully resolved and appeared as a reduction in mean unitary single-channel current amplitude. 3. This effect was voltage dependent, and on analysis was shown to be due to an open channel block by divalent ions. In the case of Mg2+, the block increased steadily with hyperpolarization. In contrast, for Ca2+ the block first increased with hyperpolarization and then decreased with further hyperpolarization beyond -70 mV, providing evidence for Ca2+ permeation of this channel. 4. This block is similar to that seen in voltage-gated calcium channels. Additionally, the cyclic nucleotide-gated channel shows some pharmacological similarities with L-type calcium channels, including a novel block of the cyclic nucleotide channel by nifedipine (50 microM). 5. Our results indicate that the sensory generator current simultaneously depends on the presence of the second messenger and on the membrane potential of the olfactory neuron.

Ambystoma↗
PubMed1992

Electrical signals in olfactory transduction.

Olfactory transduction involves a G-protein-coupled second messenger system, which results in the odor-dependent production of cAMP. The direct activation of ion channels in the cilia membrane by cAMP is the final step in producing the slow depolarization that brings the membrane potential to threshold for spike generation. Because of the central role in the transduction cascade occupied by these channels considerable effort has been directed toward understanding their behavior at a molecular level. Alternative second messenger pathways have also been proposed in olfaction, but the physiological evidence for these is less well developed.

Animals↗
PubMed1992

Neurotransmitter antagonists block some odor responses in olfactory receptor neurons.

The first step in olfactory transduction is the recognition of odor molecules by membrane bound receptors belonging to the superfamily of G-protein coupled receptors; other members of this family are involved in neurotransmission. Based on the considerable homology between individual members of this family, we have investigated the ability of well-known neurotransmitter antagonists to block the olfactory response. Adrenergic and muscarinic antagonists were found to block some odor induced currents (45-55%) with an IC50 between 15 and 75 microM. By contrast, antagonists of glutamate and GABA receptors, which do not belong to this receptor superfamily, were ineffective. These results suggest that further pharmacological analysis may be useful for characterizing and classifying the family of odor receptors.

2-Amino-5-phosphonovalerate↗
PubMed1991

Inhibition of the olfactory cyclic nucleotide gated ion channel by intracellular calcium.

When olfactory receptor neurons are exposed to sustained application of odours, the elicited ionic current is transient. This adaptation-like effect appears to require the influx of Ca2+ through the odour-sensitive conductance; in the absence of extracellular Ca2+ the current remains sustained. Odour transduction proceeds through a G-protein-based second messenger system, resulting finally in the direct activation of an ion channel by cyclic AMP. This channel is one possible site for a negative feedback loop using Ca2+ as a messenger. In recordings of single cyclic AMP gated channels from olfactory receptor neurons, the open probability of the channel in saturating cAMP concentrations was dependent on the concentration of intracellular Ca2+. It could be reduced from 0.6 in 100 nm Ca2+ to 0.09 in 3 microM Ca2+. However, as neither the single channel conductance nor the mean open time were affected by Ca+ concentration, this does not appear to be a mechanism of simple channel block. Rather, these results suggest that intracellular Ca2+ acts allosterically to stabilize a closed state of the channel.

Ambystoma↗
PubMed1991

Activation of the sensory current in salamander olfactory receptor neurons depends on a G protein-mediated cAMP second messenger system.

Olfactory receptor neurons respond to odor stimulation with an inward cationic current. Under whole-cell patch clamp, individual, isolated olfactory receptors were exposed to pharmacological agents known to interact with distinct enzymes in a putative second messenger cascade, and their response to odors was measured. IBMX prolonged the odor-evoked current and also reduced its amplitude. cAMP and cGMP induced a current electrically identical to the odor current, but the current showed desensitization only with cAMP. GTP-gamma-s prolonged and GDP-beta-s interfered with the odor-evoked current. The long latency seen in the odor response appears to be mainly due to the loading of the G protein and secondarily to the requirement for cAMP accumulation. The main source of the response decay appears to be cyclic nucleotide hydrolysis.

1-Methyl-3-isobutylxanthine↗
PubMed1991

Toward a pharmacology of odor receptors and the processing of odor images.

Odor molecules may be considered as molecular ligands which bind to receptors in the olfactory sensory neurons to give rise to the sensory response. Binding studies in whole sensory epithelia suggest that the receptors also bind muscarinic cholinergic antagonists. Preliminary electrophysiological evidence indicates that muscarinic and beta adrenergic antagonists block odor-elicited membrane currents in single isolated salamander sensory neurons. These results support the idea that models developed for analyzing ligand binding by members of the 7 transmembrane domain family of membrane receptors may apply rather closely to olfactory transduction. We suggest that sensory neurons express single receptor types with differing degrees of affinity for different ligands. We further suggest that glomeruli in the olfactory bulb function as labeled lines for particular sets of odor ligand determinants, and that interglomerular circuits bind together similar glomeruli and enhance contrast between dissimilar glomeruli. The odor image laid down in the sensory neuron population is thus subjected to abstracting and enhancement at the glomerular stage, prior to being transmitted for further processing in the deeper layers of the olfactory bulb and in the olfactory cortex.

Animals↗
PubMed1991

A kinetic model of the odor response in single olfactory receptor neurons.

The detection of odor molecules by olfactory receptors is a biochemical process, but the neural signal is electrical. The transformation of chemical information into a change in membrane potential, i.e. the process of signal transduction, is accomplished in olfactory receptor neurons by a multi-step second messenger pathway resulting finally in the activation of ion channels by cAMP. Many of the biochemical and physiological details of this process are beginning to be appreciated, giving rise to a comprehensive model of the basic mechanisms of olfactory transduction that has much in common with those of other signal transduction systems. One interesting result of these new insights is that the olfactory neuron may act more as a molecule counter than a concentration detector, as had been believed previously.

Action Potentials↗
PubMed1991

Electrotonic structure of olfactory sensory neurons analyzed by intracellular and whole cell patch techniques.

1. Experimental studies employing whole cell patch recordings from freshly isolated olfactory sensory neurons of the salamander (Ambystoma tigrinum) yield much higher estimates of specific membrane resistance (Rm) than studies using conventional intracellular recordings from in situ neurons. Because Rm is critical for understanding information transfer in these cells, we have used computational methods to analyze the possible reasons for this difference. 2. Compartmental models were constructed for both the in situ and isolated neurons, using SABER, a general-purpose simulation program. For Rm in the in situ cell, we used a high value of 100,000 omega.cm2, as estimated in the whole cell recordings from isolated cells. A shunt across the cell membrane caused by the penetrating microelectrode was simulated by several types of shunt mechanisms, and its effects on lowering the apparent value of resting membrane potential (MP), input resistance (RN), and membrane time constant (tau m) and increasing the electrotonic length (L) were analyzed. 3. A good approximation of the electrotonic properties recorded intracellularly was obtained in the in situ model with high Rm combined with an electrode shunt consisting of Na and K conductances. A raised K conductance (1-5 nS) helps to maintain the resting MP while contributing to the increased conductance, which lowers RN and shortens the apparent tau m toward the experimental values. 4. Combined shunt resistances of 0.1-0.2 G omega (5-10 nS) gave the best fits with the experimental data. These shunts were two to three orders of magnitude smaller than the values reported from intracellular penetrations in muscle cells and motoneurons. This may be correlated with the smaller electrode tips used in the recordings from these small neurons. We thus confirm the prediction that even small values of electrode shunt have relatively large effects on the recorded electrotonic properties of small neurons, because of their high RN (2-5 G omega). 5. We have further explored the effects on electrotonic structure of a nonuniform Rm by giving higher Rm values to the distally located cilia compared with the proximal soma-dendritic region, as indicated by recent experiments. For the same RN, large increases in ciliary Rm above 100,000 omega.cm2 can be balanced by relatively small decreases below that value in soma-dendritic Rm. A high ciliary Rm appears to be a specialization for transduction of the sensory input, as reported also in photoreceptors and hair cells.

Ambystoma↗