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A R Cinelli

Publications and source records attributed to A R Cinelli.

At least 19 recordsLinked to original sources

High-definition mapping of neural activity using voltage-sensitive dyes.

The distribution of patterns of activity in different brain structures has been related to the encoding and processing of sensory information. Consequently, it is important to be able to image the distribution of these patterns to understand basic brain functions. The spatial resolution of voltage-sensitive dye (VSD) methods has recently been enhanced considerably by the use of video imaging techniques. The main factor that now hampers the resolution of VSD patterns is the inherent limitation of the optical systems. Unfortunately, the intrinsic characteristics of VSD images impose important limitations that restrict the use of general deconvolution techniques. To overcomes this problem, in this study an image restoration procedure has been implemented that takes into consideration the limiting characteristics of VSD signals. This technique is based on applying a set of imaging processing steps. First, the signal-to-noise (S/N) ratio of the images was improved to avoid an increase in the noise levels during the deconvolution procedures. For this purpose, a new filter technique was implemented that yielded better results than other methods currently used in optical imaging. Second, focal plane images were deconvolved using a modification of the well-known nearest-neighbor deconvolution algorithm. But to reduce the light exposure of the preparation and simplify image acquisition procedures, adjacent image planes were modeled according to the in-focus image planes and the empirical point spread function (PSF) profiles. Third, resulting focal plane responses were processed to reduce the contribution of optical responses that originate in distant image planes. This method was found to be satisfactory under simulated and real experimental conditions. By comparing the restored and unprocessed images, it was clearly demonstrated that this method can effectively remove the out-of-focus artifacts and produce focal plane images of better quality. Evaluations of the tissue optical properties allowed assessment of the maximum practical optical section thickness using this deconvolution technique in the optical system tested. Determination of the three-dimensional PSF permitted the correct application of deconvolution algorithms and the removal of the contaminating light arising from adjacent as well as distant optical planes. The implementation of this deconvolution approach in salamander olfactory bulb allowed the detailed study of the laminar distribution of voltage-sensitive changes across the bulb layer. It is concluded that (1) this deconvolution procedure is well suited to deconvolved low-contrast images and offers important advantages over other alternatives; (2) this method can be properly used only when the tissue optical properties are first determined; (3) high levels of light scattering in the tissue reduce the optical section capabilities of this technique as well as other deconvolution procedures; and (4) use of the highest numerical aperture in the objectives is advisable because this improves not only the light-collecting efficiency to detect poor-contrast images, but also the spatial frequency differences between adjacent image planes. Under this condition it is possible to overcome some of the limitations imposed by the light scattering/birefringence of the tissue.

Algorithms↗

Flexible method to obtain high sensitivity, low-cost CCD cameras for video microscopy.

A simple method is described to extend image exposure times in video-rate CCD cameras and thereby, increase their sensitivity and reduce noise level of low-light images. Most commercial video cameras lack the capability of extending image exposures since they operate regular television timing formats. The technique described here implements the control of the exposure times by selectively gating the image readout from the CCD sensor. This prevents the cyclic clearing of photo-charges occurring at regular video-rates, allowing image integration beyond the duration of single video field periods. Image readout is controlled by the duration of external gating pulses, giving the camera an efficient operational versatility under different light conditions. This technique is applicable to standard monochrome and color CCD cameras. The evaluations described here using this technique show that the light sensitivity of an standard video-rate CCD camera can be significantly improved, generating high quality images at low-light levels. These were comparable to those obtained with image intensifiers or intensified video cameras. Cameras are still compatible with regular video equipment, since this technique preserves the normal TV synchronization signals. Results in simulated and real experimental situations confirmed that this technique enables the use of affordable video-rate CCD cameras for a variety of fluorescence microscopy and optical recording applications.

Animals↗

Regulation of Na,K-ATPase transport activity by protein kinase C.

Considerable evidence indicates that the renal Na+,K+-ATPase is regulated through phosphorylation/dephosphorylation reactions by kinases and phosphatases stimulated by hormones and second messengers. Recently, it has been reported that amino acids close to the NH2-terminal end of the Na+,K+-ATPase alpha-subunit are phosphorylated by protein kinase C (PKC) without apparent effect of this phosphorylation on Na+,K+-ATPase activity. To determine whether the alpha-subunit NH2-terminus is involved in the regulation of Na+, K+-ATPase activity by PKC, we have expressed the wild-type rodent Na+,K+-ATPase alpha-subunit and a mutant of this protein that lacks the first thirty-one amino acids at the NH2-terminal end in opossum kidney (OK) cells. Transfected cells expressed the ouabain-resistant phenotype characteristic of rodent kidney cells. The presence of the alpha-subunit NH2-terminal segment was not necessary to express the maximal Na+,K+-ATPase activity in cell membranes, and the sensitivity to ouabain and level of ouabain-sensitive Rb+-transport in intact cells were the same in cells transfected with the wild-type rodent alpha1 and the NH2-deletion mutant cDNAs. Activation of PKC by phorbol 12-myristate 13-acetate increased the Na+,K+-ATPase mediated Rb+-uptake and reduced the intracellular Na+ concentration of cells transfected with wild-type alpha1 cDNA. In contrast, these effects were not observed in cells expressing the NH2-deletion mutant of the alpha-subunit. Treatment with phorbol ester appears to affect specifically the Na+,K+-ATPase activity and no evidence was observed that other proteins involved in Na+-transport were affected. These results indicate that amino acid(s) located at the alpha-subunit NH2-terminus participate in the regulation of the Na+,K+-ATPase activity by PKC.

Animals↗

Stimulation of protein kinase C rapidly reduces intracellular Na+ concentration via activation of the Na+ pump in OK cells.

Na+ reabsorption is regulated in proximal tubules by hormones that stimulate protein kinase C (PKC). To determine whether stimulation of PKC causes a reduction in intracellular Na+ concentration ([Na+]i) that might link Na+ pump activation to increased Na+ reabsorption, [Na+]i was measured in kidney cells loaded with the Na+-sensitive fluorescent indicator SBFI. Rapid digital imaging fluorescence microscopy determinations were performed in epithelial kidney cells transfected with the rodent Na+ pump alpha1 cDNA. In 42 determinations, the basal [Na+]i was 19.7 +/- 2.4 mM. Stimulation of PKC reduced the [Na+]i to 5.6 +/- 0.6 mM in approximately 10 sec. This drastic change in [Na+]i requires a transient 74-120-fold increase in Na+ pump activity. After the new steady state [Na+]i is reached, the Na+ pump is 58% activated. The entry of Na+ into the cells is not affected by stimulation of PKC; therefore, the reduction in [Na+]i is exclusively dependent on activation of the Na+ pump. Accordingly, PKC stimulation does not affect the [Na+]i of cells expressing a mutant Na+ pump that is not stimulated by PKC. The decrease in [Na+]i observed in cells transfected with the rodent Na+ pump alpha1 cDNA is large and sufficiently fast that it is expected to stimulate rapidly passive Na+-influx into the cells, thereby accounting for the observed PKC-induced stimulation of Na+ reabsorption.

Animals↗

Salamander olfactory bulb neuronal activity observed by video rate, voltage-sensitive dye imaging. I. Characterization of the recording system.

1. In this paper we describe properties of a video imaging system used to acquire voltage-sensitive dye fluorescence signals from the salamander olfactory bulb. Sources of noise in these signals were evaluated in preparations stained with the potentiometric probe RH-414. These were compared with noise levels in signals obtained from a light-emitting diode array designed to stimulate the experimental conditions with light levels similar to those seen in the salamander bulb recordings. These experiments define a number of determinants of video image quality to standardize optical voltage measurements in the salamander olfactory bulb. 2. Images were acquired at video rates using a Newvicon camera in a standard upright microscope and digitized with an eight-bit video frame grabber. 3. Sources of noise related to camera sensitivity, stability of illumination, and mechanical vibration were characterized. Camera dark noise was less than the pixel variability due to photon noise at the camera faceplate. This pixel noise was the limiting factor for discriminating the spatial and temporal properties of the optical responses. 4. No significant noise was found to be related to image digitization, transmission, or readout by the eight-bit frame grabber. Mechanical vibration, light stability, and other sources of noise could be controlled in vitro. In this condition, voltage-sensitive dye signal noise was similar to that in stimulated experiments using the light-emitting diode array. Higher levels of noise were found in vivo; some of this was reduced by sychronizing frame acquisition to the heartbeat. 5. On the basis of photodiode and video measurements, voltage-sensitive dye responses in the salamander olfactory bulb typically fell between 0.75% and 2.5% fractional change of background fluorescence. By appropriately adjusting the video signals before analog-to-digital conversion, we could detect fractional changes of < 0.5%. 6. Both response averaging and low-bandpass spatial filtering improved the signal-to-noise ratios of the images. For small numbers of averaged runs, the best improvement was obtained by low-bandpass spatial filtering. 7. Acquisition of high-spatial resolution video images permitted the use of low-bandpass spatial filters to suppress pixel noise. The degree of spatial enhancement depended on the relationship between the size of the structures of interest, pixel density, and the properties of the convolution filter kernel. This method avoided exposure of the preparation to prolonged illumination and the necessity of applying the large numbers of repeated stimuli required for averaging.

Animals↗

Salamander olfactory bulb neuronal activity observed by video rate, voltage-sensitive dye imaging. II. Spatial and temporal properties of responses evoked by electric stimulation.

1. Video imaging of changes in voltage-sensitive dye (VSD) fluorescence was used to analyze spatial and temporal properties of activity patterns in the in vivo salamander olfactory bulb and primordium piriform cortex after electric stimulation. Distribution of activity among and within the neuronal layers was analyzed after orthodromic stimulation of the whole olfactory nerve (ON), isolated fascicles, or local epithelial sites, and after antidromic stimulation of the medial olfactory tract (OT). 2. Optical signals propagated through the bulbar layers with a sequence that correlates with electrophysiological responses. After orthodromic stimulation, VSD responses started in the glomerular layer, spread to the deeper laminae, and, after reaching the region of mitral/tufted somata, were observed as a brief burst of activity in the OT. Compound action potentials in the ON were associated with short-duration, rapidly depolarizing optical responses in the ON layer. Responses in glomerular layer and external plexiform layer (EPL) first showed in some recordings a brief, small-amplitude hyperpolarization, followed by a period of depolarization, followed by a second, longer-lasting hyperpolarization. The periods of optical hyperpolarization could be related to events observed in intracellular mitral/tufted cell recordings. 3. With shocks delivered to the entire ON, depolarizing responses were nonhomogeneously distributed, appearing as multiple foci or bands of activity. Spatial patterns within each bulbar layer had poorly defined borders. Sites showing short-latency responses were often those with the largest and longest-lasting activity. 4. Increasing the intensity of stimulation to the ON enhanced the size and duration of the depolarizing and hyperpolarizing responses. The short-latency, early hyperpolarization was best seen with low-intensity, peripherally placed stimuli. 5. ON stimulation also elicited activity in the contralateral bulb. Activity started at the innermost layers and spread in patches to regions of the EPL just beneath the glomeruli. These had durations similar to ipsilateral responses, but longer latencies. A period of early hyperpolarization, longer than that on the ipsilateral side, was followed by prolonged depolarization and then by a second, later hyperpolarization. 6. Antidromic stimuli applied to the OT evoked optical responses consisting of a period of depolarization followed by hyperpolarization, similar to the components elicited by orthodromic stimuli. These responses had short time courses, began in the deeper layers, and spread to the superficial region of the bulb usually without reaching the glomerular region. 7. Punctate stimulation of the mucosa or nerve elicited depolarizing and hyperpolarizing events that depended on the stimulation site.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Salamander olfactory bulb neuronal activity observed by video rate, voltage-sensitive dye imaging. III. Spatial and temporal properties of responses evoked by odorant stimulation.

1. Activity patterns across and within the laminae of the olfactory bulb were analyzed by imaging voltage-sensitive dye responses during odorant stimulation of all or part of the ventral olfactory mucosa. 2. The time course of the signals was generally characterized by a brief, small hyperpolarization, followed by a period of depolarization, and then a longer-lasting hyperpolarization similar to that seen with electric stimulation but with longer durations. 3. The activity was distributed nonhomogeneously across the bulbar laminae in the form of spatially segregated clusters having bandlike appearances. Clusters were observed with three monomolecular odorants, amyl acetate, ethyl-n-butyrate, and limonene, and with the complex odor of meal worms. Although response patterns to different odorants overlapped, they also showed differences in overall distribution. 4. Delivery of high odorant concentrations increased the size of the activated areas and accentuated the degree of response pattern overlap among different odorants. The general properties of the response patterns generated by each odorant were, however, similar at different odorant concentrations and in each of the animals tested. 5. The spatial and temporal distributions of the bulbar responses were somewhat similar regardless of whether the odorants were applied to local epithelial regions via punctate stimulation or to the entire mucosa. Certain regions did, however, have lower thresholds than others for eliciting bulbar activity in response to particular odorants. 6. Odorants applied to regions of the epithelium outside the areas of maximum sensitivity elicited odorant-related activity patterns with depolarizing and hyperpolarizing components similar to those seen with overall stimulation, but only if higher concentrations were used. Activation of distributed odorant sensitivities presumably gave rise to these patterns. 7. These data suggest that subsets of odorant receptor types are found in different areas of the olfactory epithelium, and demonstrate that there is widespread distribution across the epithelium of receptors sensitive to particular odorants. On the basis of the structure of these epithelial fields and the bulb response patterns that they relate to, these findings also provide evidence for complex spatial relationships between the olfactory epithelium and bulb. 8. The findings from this study suggest that representation of odor information in the salamander olfactory bulb does not occur by activation of a few selective bulbar regions, each related to a different odorant species. Instead, large regions of bulbar circuitry are involved in which molecular epitopes may be the unit of representation. Incorporation of new data presented here into a hypothesis of odor coding is discussed.

Animals↗

Are there structural and functional modules in the vertebrate olfactory bulb?

A number of different recording methods have shown that odorants elicit patterns of neuronal activity widely distributed across cells of the olfactory receptor epithelium, olfactory bulb, and piriform cortex in the vertebrate olfactory system. These findings suggest that the physicochemical properties of odorant molecules are processed by distributed coding mechanisms activated in parallel in olfactory circuits in order to characterize a single, "monomolecular" odorant. These findings also suggest that the response patterns seen at higher levels are set up by differential responses in peripheral receptor cells of the olfactory epithelium. One requirement for understanding the details of this proposed encoding scheme is correlation of odor-generated patterns with the components of these circuits. In this paper, results from 2-deoxyglucose and voltage-sensitive dye studies suggest that certain components of these responses may relate to patterns established in reproducibly identifiable aggregates of bulbar cells. These findings are consistent with previous observations suggesting that columnar groups of periglomerular, mitral/tufted and granule cells, oriented perpendicular to the laminae of the bulb, are functionally related to one another. Such cell groups or modules, when activated in parallel, could serve as building block components of the complete ensemble response. According to this hypothesis, different sets of such modules would be activated with different odorant stimuli and modules could be shared to the degree to which the physicochemical properties of the different stimuli overlap.

Animals↗

Intracellular injection of vital dyes into single cells in the salamander olfactory epithelium.

Intracellular vital dye injection was used to examine the morphology of single sustentacular and receptor cells and the developmental fate of individual basal cells in the olfactory epithelium of the tiger salamander. In acute experiments, Lucifer yellow injections were used to identify single basal, receptor or sustentacular cells on the basis of their overall morphology. Dye-coupling between a number of the different epithelial cells was observed. Progeny of basal cells were examined by following labeled cells for up to 2 weeks using intracellular injection of rhodamine-labeled dextran. These experiments indicate that some olfactory epithelial cells are dye-coupled and that dye-filled basal cells can undergo division and migration.

Ambystoma↗

Dendritic origin of late events in optical recordings from salamander olfactory bulb.

1. Optical recordings of membrane-potential changes were used to characterize the origin and properties of the electrical signals from the dendritic level in slices of the salamander olfactory bulb. 2. The optical events were correlated with field-potential waves recorded simultaneously. Both responses exhibited patterns similar to those found in other species. 3. Orthodromic stimulation evoked a compound action potential in the olfactory nerve fibers, followed by two additional principal waves (N1 and N2). These field-potential waves reflected excitatory postsynaptic potentials at the primary mitral/tufted and granule cell dendrites, respectively. 4. Extrinsic optical signals from horizontal slices stained with the pyrazo-oxonal dye RH-155 showed a characteristic sequence of depolarizing and hyperpolarizing events. All of the signals exhibited a wavelength dependence expected for this dye and were abolished in the presence of high K+ in the bath. 5. According to their time courses, depolarizing responses under normal recording conditions were divided into two components, fast and slow. Orthodromic stimuli evoked a fast presynaptic response that represents synchronous compound action potentials from olfactory nerve fibers. At subglomerular levels, additional fast responses could often be recorded at the peri/subglomerular level and in the mitral/tufted somata region. These postsynaptic responses partially coincided with the rising phase of a different depolarizing signal, a slow component characterized by its prolonged time course. 6. With orthodromic stimulation, this slow signal attained its largest amplitude in the zone between the glomeruli and the superficial part of the external plexiform layer (EPL). Antidromic stimuli evoked a signal with some similarities to the one evoked orthodromically, but originating in deeper EPL regions. 7. Slow components were characterized by their Ca dependence. Low Ca2+ medium, or calcium channel blockers, suppressed this optical component, whether evoked orthodromically, antidromically, or by direct stimulation. In addition, Ba2+ (2.5-3.6 mM) in the bath did not abolish these responses, suggesting that they do not reflect a glial depolarization in response to elevated extracellular K+ concentration ([K+]o). 8. Locally applied stimuli next to the glomerular layer elicited these signals in 5-10 microM tetrodotoxin (TTX) or in low extracellular Na+ concentration ([Na+]o) medium, but antidromic or orthodromic stimuli failed to evoke the response under these conditions. The sizes of the responses to local stimuli remained constant, but an increase in their duration was observed in either TTX or low [Na+]o. 9. gamma-Aminobutyric acid (GABA) and baclofen reduced the size of the slow components in a dose-dependent manner.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

Multiple site optical recording of transmembrane voltage (MSORTV), single-unit recordings, and evoked field potentials from the olfactory bulb of skate (Raja erinacea).

1. Multiple site optical recording of transmembrane voltage (MSORTV), together with conventional extracellular electrophysiological techniques were utilized with in vivo and in vitro preparations of the olfactory bulb of the Atlantic skate Raja erinacea to analyze electrical activity simultaneously in layers deep to the glomerular layer. 2. In the living animals and the in vitro isolated olfactory bulb, orthodromic stimulation evoked a compound action potential in the olfactory nerve fibers, followed by a series of early field-potential waves (N1, P1, N2, P2, N3, and N4). During paired stimulation experiments, unusual patterns of facilitation and suppression were observed for the N2 wave. 3. After orthodromic stimulation, single units, presumably mitral/tufted cells, exhibited a period of early discharge, followed by a period of suppression of spontaneous activity and of their test response in a pair stimulation paradigm. Some neurons also exhibited a labile period of reexcitation that was accompanied by a late surface negative field potential; these responses were also present in olfactory bulb slices. 4. Extrinsic absorption changes obtained from 500-microns saggital slices of the olfactory bulb, stained with the pyrazooxonal dye RH-155, consisted mainly of two types of depolarizing responses, a fast and a slow component, followed under some conditions by a late hyperpolarization. All signals exhibited wavelength dependences typical of the action spectrum of RH-155 and were abolished in the presence of tetrodotoxin (TTX) or high K+ in the bath. 5. The fast component of the optical signal represents synchronous compound action potentials conducted by the olfactory nerve fibers or evoked in the mitral/tufted somata and axonal pathways. The slow depolarizing optical signal appeared, after orthodromic stimuli, mainly in the zone between the glomeruli and the mitral/tufted layer; barium (1-10 mM), which depolarizes glial cells, increased its size and duration, suggesting that this signal does not reflect a glial response to [K+]o. 6. Different condition/test (C/T) intervals produced partial or complete suppression of the test response, depending on the recording site and the stimulus intensity. Just threshold orthodromic stimuli evoked an intermediate period of facilitation of the slow signals. A similar period was also observed in the N2 wave of the field potential. 7. Calcium channel blockers such as cadmium ion, or a low Ca2+ medium, suppressed the slow optical component whether evoked by orthodromic, antidromic, or direct stimulation. gamma-Aminobutyric acid (GABA) and baclofen also reduced or blocked the slow component of the extrinsic absorption signal.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Current generators and properties of late components evoked in rat olfactory cortex.

Following main olfactory bulb (MOB) stimulation at frequencies of 0.1-0.3 Hz, in addition to early field potentials, a frequency-sensitive, surface negative late N2 wave (latency range: 63-96 msec) followed occasionally by a late N3 transient, was evoked in the piriform cortex and endopiriform nucleus of the rat. The N2 wave inverted polarity at the Ib-II cortical layer interface (P2 wave) and was associated with late unit discharges 200 to 1200 microns deep to the turnover point. Response probability, peak latency, recovery curve and frequency-sensitivity of the P2 wave were not significantly different in animals under urethane or pentobarbital. Current-source-density (CSD) analysis revealed that the N2 wave generators were localized to the Ib-II layer interface. Since inhibitory activity does not contribute substantially to the second derivative curve, CSD analysis strengthens the assumption that late components (LCs) are excitatory events (compound EPSPs) presumably generated on the proximal apical dendritic segments of pyramidal cells by association axons. The early "b" wave in a test response was facilitated, rather than occluded, when a LC was present in the conditioning response, or when the priming volley was delivered to the mediodorsal thalamic nucleus. Clustering of unit and field activity in two distinct periods of the evoked response separated by a prolonged interval of cell silence suggests that cortical coding of olfactory cues might be more efficiently achieved by temporal modulation of the neuronal response rather than by spatial distribution of firing patterns.

Animals↗

A computer program for automatic plotting of isopotential contours in CNS.

A computer program was developed in a Basic (Applesoft) version for generating up to five isopotential curves from field potentials recorded in nervous structures. Voltages are fed according to a cartesian coordinate system, and an area is delineated each four points in which a certain number of intermediate voltage points are calculated, according to the required resolution. The calculated values are compared to those prefixed for each curve and, if similar, their coordinates are stored in corresponding bidimensional matrixes. A special subroutine was designed for constructing an isometric tridimensional perspective of the isopotential curve ensemble. The reliability of this program was tested in the localization of sensory representation areas on the neocortex of the South American armadillo (Chaetophractus vellerosus) studied by evoked potential mapping following visual, auditory and somatosensory stimuli. The isopotential curves traced permitted a quantitative evaluation of the cortical activated areas, and from their topographical distribution, relative unresponsive zones could be inferred where only inconspicuous responses were obtained. It is concluded that the present program provides a reliable and fast method for studying the evoked potential's spatial distribution over the entire neocortex. In addition, it can be extended to the study of curves or contours which connect equivalent values pertaining to biophysical magnitudes other than voltage data.

Animals↗

Reciprocal functional connections of the olfactory bulbs and other olfactory related areas with the prefrontal cortex.

Reciprocal putative connections of the prefrontal cortex (PFC) (agranular insular, ventral and lateral orbital region) with the ipsi and contralateral main olfactory bulb (IOB; COB), the mediodorsal thalamic nucleus (MD), the basolateral amygdaloid nucleus (BLA) and the piriform cortex (PC) were investigated with electrophysiological techniques. Evoked field responses and orthodromic unit driving, generated in PFC following electrical stimulation of the above mentioned structures, were abolished following topical application of KCl, except for COB evoked mass potentials. Thus, locally generated activity was elicited in agranular insular cortex following IOB activation, the same region where recently, the taste cortex in the rat was localized. Since gustatory-visceral afferent information reaches insular cortex via 2-3 synaptic relays, autonomic, olfactory and gustatory inputs may interact at this level, and, as suggested previously for the mouse, play a key integrative role in flavor perception. Antidromically invaded neurons, 47% of which were identified by the collision-extinction technique, were also found in PFC areas which overlapped to a considerable extent with those from which orthodromic unit responses were obtained. In particular, closely spaced neurons in ventrolateral orbital (VLO) and lateral orbital (LO) regions were antidromically invaded following IOB and PC shocks; some neurons antidromically discharged by IOB were also transsynaptically activated following PC stimulation. These findings are in agreement with recent neuroanatomical studies which demonstrate axonal projections from PFC neurons to the IOB and COB in the rat and South American armadillo. In addition, stimulation of PFC regions dorsal to the rhinal fissure mostly inhibited spontaneous unit discharges recorded at the mitral cell layer of the IOB, suggesting that this effect may be partially mediated by excitatory inputs of prefrontal axons onto granule cells. The conduction properties, antidromic thresholds and activity-dependent variations in conduction velocity (CV) of bulbopetal neurons in prefrontal cortex were found to be similar to those exhibited by cells projecting to the IOB from olfactory peduncle regions, but not to those present in bulbopetal neurons of the horizontal limb of diagonal band, indicating that the OB may be subjected to centrifugal control by at least two cell groups differing in both histochemical and electrophysiological properties.

Afferent Pathways↗

Axonal projections and conduction properties of olfactory peduncle neurons in the armadillo (Chaetophractus vellerosus).

Extracellular unit recording was employed to study the axonal properties and efferent projections of antidromically identified neurons in the olfactory peduncle (OP) region of a primitive eutherian macrosmatic mammal, the south american armadillo (Chaetophractus vellerosus). Of 72 cells which satisfied the criteria for antidromic invasion, 55 (76%) and 17 (24%) responded to ipsi- and contralateral olfactory bulb (IOB; COB) stimulation, respectively. The absolute refractory period (3.25 +/- 0.3 ms; mean +/- SE) and the conduction velocity (CV; 1.94 +/- 0.2 m/s; mean +/- SE) of IOB and COB driven neurons were negatively correlated (r = -0.52; p less than 0.001). In paired-shock tests (8-1950 ms interval), and early supernormal period (SPN) of increased CV and excitability was found following the relative refractory period in 82% of tested cells (N = 50); this period was followed by a late subnormal phase (SBN) of decreased CV and increased threshold in 58% of neurons (N = 50). Significant correlations were found to exist between: CV and absolute magnitude of latency variation (r = -0.55; p less than 0.001; n = 43), CV and duration of SPN and SBN periods (r = -0.60; p less than 0.002; n = 24 and r = 0.58; p less than 0.02; n = 19, respectively) and between duration of SPN and SBN phases (r = 0.79; p less than 0.001; n = 30). Maximum latency variation during the SPN and SBN periods was attained in a gradual, additive manner.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

An automatic device for determining threshold variations in antidromically activated neurons.

A device was designed and constructed with the purpose of evaluating threshold variations for antidromic invasion of extracellularly recorded neurons. Identification of a neuron is carried out by two procedures, an amplitude discriminator, which isolates the spike from the baseline noise, and by a latency window which is set accordingly to the neuron's antidromic latency. During threshold evaluation, the duration of an electric pulse applied to the neuron's axon is automatically varied depending on the presence or not of an action potential. For a given spike, the stimulus is progressively decreased (-delta i) up to a point where the neuron ceases to respond and thereafter, the stimulus amplitude is progressively increased (+delta i) until slightly suprathreshold values are obtained. The procedure guarantees a discharge probability of the neuron equivalent to 50% of all applied stimuli, and the simple monitoring of the stimulus amplitude is enough to obtain the threshold value for a predetermined intensity. The reliability of this device was checked in studies related to threshold variations in neurons antidromically driven in prefrontal cortex following stimulation of the ipsi and contralateral olfactory bulb. Variations in excitability were found during and following tetanic stimulation and throughout the axon's supernormal conduction period. This technique allows the assessment of threshold variations in antidromic driving, not only in the present experimental design, but also in other conditions induced by changes in extracellular ionic concentrations, drug applications or in those produced by excitatory or inhibitory synaptic activity on the neuron under study.

Animals↗

Variations in axonal conduction velocity in olfactory peduncle neurons of the armadillo.

Following electrical stimulation of the ipsi- or contralateral olfactory bulb, antidromically invaded neurons in the olfactory peduncle (OP) of the armadillo (a primitive mammal with a low brain temperature) show progressive decreases in conduction velocity when challenged with stimulation frequencies of 1-40 Hz. Antidromic latency also decreased or increased in a gradual, additive manner during the super- and subnormal period following twin pulses at intervals of 8-2000 ms. Since centrifugal OP axons excite inhibitory granular cells in the olfactory bulb, these effects may bear on the problem of the control of mitral cell excitability by central structures.

Animals↗