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M Wachowiak

Publications and source records attributed to M Wachowiak.

14 recordsLinked to original sources
PubMed2001

Representation of odorants by receptor neuron input to the mouse olfactory bulb.

To visualize odorant representations by receptor neuron input to the mouse olfactory bulb, we loaded receptor neurons with calcium-sensitive dye and imaged odorant-evoked responses from their axon terminals. Fluorescence increases reflected activation of receptor neuron populations converging onto individual glomeruli. We report several findings. First, five glomeruli were identifiable across animals based on their location and odorant responsiveness; all five showed complex response specificities. Second, maps of input were chemotopically organized at near-threshold concentrations but, at moderate concentrations, involved many widely distributed glomeruli. Third, the dynamic range of input to a glomerulus was greater than that reported for individual receptor neurons. Finally, odorant activation slopes could differ across glomeruli, and for different odorants activating the same glomerulus. These results imply a high degree of complexity in odorant representations at the level of olfactory bulb input.

Acetates↗
PubMed2001

Patch-clamp analysis of voltage-activated and chemically activated currents in the vomeronasal organ of Sternotherus odoratus (stinkpot/musk turtle).

The electrophysiological basis of chemical communication in the specialized olfactory division of the vomeronasal (VN) organ is poorly understood. In total, 198 patch-clamp recordings were made from 42 animals (Sternotherus odoratus, the stinkpot/musk turtle) to study the electrically and chemically activated properties of VN neurons. The introduction of tetramethylrhodamine-conjugated dextran into the VN orifice permitted good visualization of the vomeronasal neural epithelium prior to dissociating it into single neurons. Basic electrical properties of the neurons were measured (resting potential, -54.5 +/- 2.7 mV, N=11; input resistance, 6.7 +/- 1.4 G Omega, N=25; capacitance, 4.2 +/- 0.3 pF, N=22; means +/- S.E.M.). The voltage-gated K(+) current inactivation rate was significantly slower in VN neurons from males than in those from females, and K(+) currents in males were less sensitive (greater K(i)) to tetraethylammonium. Vomeronasal neurons were held at a holding potential of -60 mV and tested for their response to five natural chemicals, female urine, male urine, female musk, male musk and catfish extract. Of the 90 VN neurons tested, 33 (34 %) responded to at least one of the five compounds. The peak amplitude of chemically evoked currents ranged from 4 to 180 pA, with two-thirds of responses less than 25 pA. Urine-evoked currents were of either polarity, whereas musk and catfish extract always elicited only inward currents. Urine applied to neurons harvested from female animals evoked currents that were 2-3 times larger than those elicited from male neurons. Musk-evoked inward currents were three times the magnitude of urine- or catfish-extract-evoked inward currents. The calculated breadth of responsiveness for neurons presented with this array of five chemicals indicated that the mean response spectrum of the VN neurons is narrow (H metric 0.11). This patch-clamp study indicates that VN neurons exhibit sexual dimorphism in function and specificity in response to complex natural chemicals.iol

Animals↗
PubMed2000

Odors elicit three different oscillations in the turtle olfactory bulb.

We measured the spatiotemporal aspects of the odor-induced population response in the turtle olfactory bulb using a voltage-sensitive dye, RH414, and a 464-element photodiode array. In contrast with previous studies of population activity using local field potential recordings, we distinguished four signals in the response. The one called DC covered almost the entire area of the olfactory bulb; in addition, three oscillations, named rostral, middle, and caudal according to their locations, occurred over broad regions of the bulb. In a typical odor-induced response, the DC signal appeared almost immediately after the start of the stimulus, followed by the middle oscillation, the rostral oscillation, and last, the caudal oscillation. The initial frequencies of the three oscillations were 14.1, 13.0, and 6.6 Hz, respectively. When the rostral and caudal oscillations occurred together, their frequencies differed by a factor of 1.99 +/- 0.01. The following evidence suggests that the four signals are functionally independent: (1) in different animals some signals could be easily detected whereas others were undetectable; (2) the four signals had different latencies and frequencies; (3) the signals occurred in different locations and propagated in different directions; (4) the signals responded differently to changes in odor concentration; (5) the signals had different shapes; and (6) the rostral and caudal signals added in a simple, linear manner in regions where the location of the two signals overlapped. However, the finding that the frequency of the rostral oscillation is precisely two times that of the caudal oscillation suggests a significant relationship between the two. The location of the caudal oscillation in the bulb changed from cycle to cycle, implying that different groups of neurons are active in different cycles. This result is consistent with the earlier findings in the olfactory system of the locust (). Our results suggest an additional complexity of parallel processing of olfactory input by multiple functional population domains.

Animals↗
PubMed2000

A computer analysis of reflex eyelid motion in normal subjects and in facial neuropathy.

OBJECTIVE: To demonstrate how computerized eyelid motion analysis can quantify the human reflex blink. DESIGN: Seventeen normal subjects and 10 patients with unilateral facial nerve paralysis were analyzed. BACKGROUND: Eyelid closure is currently evaluated by systems primarily designed to assess lower/midfacial movements. The methods are subjective, difficult to reproduce, and measure only volitional closure. Reflex closure is responsible for eye hydration, and its evaluation demands dynamic analysis. METHODS: A 60Hz video camera incorporated into a helmet was used to analyze blinking. Reflective markers on the forehead and eyelids allowed for the dynamic measurement of the reflex blink. Eyelid displacement, velocity and acceleration were calculated. The degree of synchrony between bilateral blinks was also determined. RESULTS: This study demonstrates that video motion analysis can describe normal and altered eyelid motions in a quantifiable manner. CONCLUSIONS: To our knowledge, this is the first study to measure dynamic reflex blinks. Eyelid closure may now be evaluated in kinematic terms. This technique could increase understanding of eyelid motion and permit more accurate evaluation of eyelid function. Dynamic eyelid evaluation has immediate applications in the treatment of facial palsy affecting the reflex blink. Relevance No method has been developed that objectively quantifies dynamic eyelid closure. Methods currently in use evaluate only volitional eyelid closure, and are based on direct and indirect observer assessments. These methods are subjective and are incapable of analyzing dynamic eyelid movements, which are critical to maintenance of corneal hydration and comfort. A system that quantifies eyelid kinematics can provide a functional analysis of blink disorders and an objective evaluation of their treatment(s).

Acceleration↗
PubMed2000

Imaging membrane potential with voltage-sensitive dyes.

Membrane potential can be measured optically using a variety of molecular probes. These measurements can be useful in studying function at the level of an individual cell, for determining how groups of neurons generate a behavior, and for studying the correlated behavior of populations of neurons. Examples of the three kinds of measurements are presented. The signals obtained from these measurements are generally small. Methodological considerations necessary to optimize the resulting signal-to-noise ratio are discussed.

Animals↗
PubMed1999

Presynaptic inhibition of primary olfactory afferents mediated by different mechanisms in lobster and turtle.

Presynaptic regulation of transmission at the first olfactory synapse was investigated by selectively imaging axon terminals of receptor neurons in the lobster olfactory lobe and turtle olfactory bulb. In both species, action potential propagation into axon terminals after olfactory nerve stimulation was measured using voltage-sensitive dyes. In addition, in the turtle, calcium influx into terminals was measured by selectively labeling receptor neurons with dextran-conjugated calcium indicator dyes. In the lobster, application of the inhibitory transmitters GABA or histamine suppressed action potentials in the terminals. The suppression was blocked by picrotoxin and cimetidine, respective antagonists to lobster GABA and histamine receptors. These results suggest that previously characterized GABA and histaminergic interneurons regulate olfactory input by suppressing action potential propagation into axon terminals of olfactory afferents. In contrast, in the turtle olfactory bulb, neither GABA nor dopamine had any effect on receptor cell action potentials as measured with voltage-sensitive dyes. However, calcium influx into axon terminals was reduced by the GABA(B) agonist baclofen and the dopamine D(2) agonist quinpirole, and paired-pulse suppression of calcium influx was reduced by the GABA(B) antagonist saclofen. These results indicate that in the turtle, GABA and dopamine mediate presynaptic inhibition not by affecting action potentials directly, as in the lobster, but by reducing calcium influx via GABA(B) and dopamine D(2) receptors. Thus, although mediated by different cellular mechanisms, presynaptic regulation of olfactory input to the CNS, via dual synaptic pathways, is a feature common to vertebrates and invertebrates. This inhibition may be important in the processing of olfactory information.

Animals↗
PubMed1998

The initiation and sequence of digital joint motion. A three-dimensional motion analysis.

We studied the initiation and sequence of digital joint motion during unrestricted flexion and extension using a 3-D motion analysis of all fingers moving simultaneously. Our results showed that motion started in a single joint in 83%, of flexion and 80%, of extension cycles. The DIP joint initiated flexion and extension in the index, middle, and ring fingers, but in the little finger, flexion started in the PIP joint, and extension in the MP joint. The two most frequent sequences of joint movement during flexion of the three radial fingers were DIP-PIP-MP and PIP-DIP-MP. The two most frequent sequences during extension of the three radial fingers were DIP-MP-PIP followed by DIP-MP/PIP. In the little finger, however, the most frequent sequences during flexion were PIP-DIP-MP followed by DIP-PIP-MP and during extension, DIP-MP/PIP followed by PIP/DIP-MP.

Adult↗
PubMed1998

Presynaptic afferent inhibition of lobster olfactory receptor cells: reduced action-potential propagation into axon terminals.

Action-potential propagation into the axon terminals of olfactory receptor cells was measured with the use of voltage-sensitive dye imaging in the isolated spiny lobster brain. Conditioning shocks to the olfactory nerve, known to cause long-lasting suppression of olfactory lobe neurons, allowed the selective imaging of activity in receptor cell axon terminals. In normal saline the optical signal from axon terminals evoked by a test stimulus was brief (40 ms) and small in amplitude. In the presence of low-Ca2+/high-Mg2+ saline designed to reduce synaptic transmission, the test response was unchanged in time course but increased significantly in amplitude (57 +/- 16%, means +/- SE). This increase suggests that propagation into receptor cell axon terminals is normally suppressed after a conditioning shock; this suppression is presumably synaptically mediated. Thus our results show that presynaptic inhibition occurs at the first synapse in the olfactory pathway and that the inhibition is mediated, at least in part, via suppression of action-potential propagation into the presynaptic terminal.

Action Potentials↗
PubMed1997

Ionotropic GABA receptor from lobster olfactory projection neurons.

This study reports an ionotropic GABA (gamma-aminobutyric acid) receptor in projection neurons acutely dissociated from the olfactory lobe of the brain of the spiny lobster and analyzed by whole cell and cell-free patch-clamp recording. GABA evokes a macroscopic current in the cells that is linear from -100 to + 100 mV, reverses at the imposed chloride equilibrium potential, has a permeability sequence of Cl- > acetate > bicarbonate > phosphate > propionate and SCN- > Br- > I- > Cl- > F-, and is reversibly blocked by the Cl channel blocker picrotoxin but not tert-butylbicyclophosphorothionate (TBPS). The current is bicuculline insensitive and activated by muscimol, isoguvacine, cis-4-aminocrotonic acid (CACA), and trans-aminocrotonic acid (TACA), as well as by the GABA(C)-receptor antagonists 4,5,6,7-tetrahydroisoxazolo [5,4,-c]pyridin-3-ol (THIP), 3-amino-1-propanesulfonic acid (3-APS), and imidazole-4-acetic acid (I-4AA), but not the GABA(B)-receptor agonists baclofen and 3-aminopropylphosphonic acid (3-APA). Agonist potency for the receptor is TACA > muscimol > GABA > I-4AA > isoguvacine > 3-APS > CACA > THIP. Unitary chloride currents in cell-free, outside-out patches from the cells share enough of these pharmacological properties to indicate that the channel underlies the macroscopic current. The receptor mediates an inhibitory current in the cells in vivo. The receptor is similar, if not identical, to one from neurons cultured from the thoracic ganglia of the clawed lobster. The more extensive pharmacological characterization of the receptor reported here indicates that this lobster CNS receptor is pharmacologically distinct from previously characterized ionotropic GABA receptors.

Animals↗
PubMed1997

Local interneurons define functionally distinct regions within lobster olfactory glomeruli

Whole-cell recording coupled with biocytin injection revealed four types of interneurons intrinsic to the olfactory lobe (OL) of the spiny lobster Panulirus argus. Each type of neuron had a distinct pattern of arborization within the three anatomically defined regions of OL glomeruli (cap, subcap and base). Type I interneurons innervated all three regions, while types II, III and IV branched only in the cap, subcap and base, respectively. Type I interneurons responded to electrical stimulation of the antennular (olfactory) nerve with a burst of 1­20 action potentials and a 1­10 s depolarization. Type II (cap) interneurons responded to the same input with a burst of 1­3 action potentials followed by a shorter hyperpolarization. Type III (subcap) interneurons responded with a burst of 1­6 action potentials followed by a delayed, 0.5­4 s depolarization. Type IV (base) interneurons responded with a brief depolarization or a burst of 1­3 action potentials followed by a 1 s hyperpolarization. The regionalized arborization and the different response properties of the type II, III and IV interneurons strongly imply that lobster olfactory glomeruli contain functionally distinct regions, a feature that should be useful in understanding the multiple synaptic pathways involved in processing olfactory input.

Journal Article↗
PubMed1994

Enhancement of gustatory nerve fibers to NaCl and formation of ion channels by commercial novobiocin.

Single fibers of the rat chorda tympani nerve were used to study the mechanism of action of the antibiotic novobiocin on salt taste transduction. In the rat, novobiocin selectively enhanced the responses of sodium-specific and amiloride-sensitive chorda tympani nerve fibers (N type) without affecting more broadly responsive cation-sensitive and amiloride-insensitive fibers (E type). In the presence of amiloride, novobiocin was ineffective at enhancing the response of N-type fibers toward sodium chloride. Novobiocin also increased the conductance of bilayers formed from neutral lipids by forming nonrectifying ion channels with low conductance (approximately 7 pS in 110 mM NaCl), long open times (several seconds and longer), and high cation selectivity. Amiloride did not alter either the conductance or kinetics of these novobiocin channels. These observations suggest that even though novobiocin is able to form cation channels in lipid bilayers, and possibly in cell membranes as well, its action on the salt-taste response is through modulation of existing amiloride-sensitive sodium channels.

Amiloride↗
PubMed1993

Stereoselective detection of amino acids by lobster olfactory receptor neurons.

1. Biochemical and electrophysiological assays were used to test the hypothesis that the olfactory system of the Caribbean spiny lobster, Panulirus argus, contains populations of chemosensory receptors that are differentially sensitive to the L- and D-stereoisomers of the amino acid alanine. 2. Independent binding sites for L-alanine (dissociation constant (KD) of 6.6 microM and maximum binding (Bmax) of 16.8 fmole/microgram protein) and for D-alanine (KD of 21.6 microM and Bmax of 17.8 fmole/microgram protein) were characterized biochemically. The interaction of ligand with each binding site is rapid, reversible and saturable with respect to both time and concentration. 3. Based on a difference of at least 20% in the relative sensitivity of an olfactory receptor cell to alanine enantiomers, 44% and 34% of the 77 neurons tested were classified as L-alanine and D-alanine sensitive, respectively. The relative sensitivity to alanine enantiomers was independent of the concentration tested. Stereoselective receptors are likely for 17 of 20 other amino acids tested. 4. The congruence of biochemical and electrophysiological results leads to the conclusion that the lobster's responses to D- and L-alanine are mediated by receptors specific for each stereoisomer and that the receptors are differentially distributed among receptor cells.

Alanine↗