Detection of tactile inputs in the rat vibrissa pathway

The rapid detection of sensory inputs is crucial for survival. Sensory detection explicitly requires the integration of incoming sensory information and the ability to distinguish between relevant information and ongoing neural activity. In this study, head-fixed rats were trained to detect the presence of a brief deflection of their whiskers resulting from a focused puff of air. The animals showed a monotonic increase in response probability and a decrease in reaction time with increased stimulus strength. High-speed video analysis of whisker motion revealed that animals were more likely to detect the stimulus during periods of reduced self-induced motion of the whiskers, thereby allowing the stimulus-induced whisker motion to exceed the ongoing noise. In parallel, we used voltage-sensitive dye (VSD) imaging of barrel cortex in anesthetized rats receiving the same stimulus set as those in the behavioral portion of this study to assess candidate codes that make use of the full spatiotemporal representation and to compare variability in the trial-by-trial nature of the cortical response and the corresponding variability in the behavioral response. By application of an accumulating evidence framework to the population cortical activity measured in separate animals, a strong correspondence was made between the behavioral output and the neural signaling, in terms of both the response probabilities and the reaction times. Taken together, the results here provide evidence for detection performance that is strongly reliant on the relative strength of signal versus noise, with strong correspondence between behavior and parallel electrophysiological findings.     

The development of whisker control in rats in relation to locomotion

Adult rats sweep their large facial whiskers (macrovibrissae) back and forth in a rhythmic pattern known as "whisking". Here we examine how these whisker movements develop in relation to other aspects of exploratory behavior, particularly locomotion. We analyzed 963 high-speed video recordings of neonatal rats, from P1 (Post-natal day 1) to P21, and measured the emergence of whisker control and of head, body, and limb movements. Prior to P11, whisker movements were largely limited to unilateral retractions accompanying head turns. Between P11 and P13 bilateral whisking emerged alongside increased forward locomotion and improved control of the head. Contact-induced modulations of whisking symmetry, synchrony, and whisker spread emerge shortly thereafter but continue to develop until at least P18, coinciding with the emergence of adult-like locomotion patterns such as rearing. Overall, whisking develops alongside increasing locomotor competence indicating that active vibrissal sensing plays an important role in the exploratory behavior of the developing animal.     

Rapid plasticity follows whisker pairing in barrel cortex of the awake rat

Synaptic plasticity can be induced easily throughout life in the rodent somatic sensory cortex. Trimming all but two whiskers on one side of an adult rat’s face, called ‘whisker pairing’, causes the active (intact) whiskers to develop a stronger drive on cortical cells in their respective barrel columns, while inactive (trimmed) whisker efficacy is down-regulated. To date, this type of activity-dependent plasticity has been induced by trimming all but two whiskers, letting the rats explore their environment from 1 day to 1 month, after which cortical responses were analyzed physiologically under anesthesia. Such studies have enhanced our understanding of cortical plasticity, but the anesthesia complicates the examination of changes that occur in the first few hours after whisker trimming. Here we assayed the short-term changes that occur in alert, active animals over a period of hours after whisker trimming. The magnitude of barrel cortex evoked responses was measured in response to stimulation of the cut and paired whiskers of rats under several conditions: (a) whisking in air (control), (b) active whisking of an object by the rat, and (c) epochs of passive whisker stimulation to identify the onset of whisker pairing plasticity changes in cortex. The main difference between whisking in air without contact and passive whisker stimulation is that the former condition induces an increased response to stimulation of inactive cut whiskers, while the latter condition increases the responses to the stimulated whiskers. The results support the conclusion that whisker pairing plasticity in barrel cortex occurs within 4 h after whisker trimming in an awake, alert animal.     

Protracted development of responses to whisker deflection in rat trigeminal ganglion neurons

The rodent whisker-to-barrel pathway constitutes a major model system for studying experience-dependent brain development. Yet little is known about responses of neurons to whisker stimulation in young animals. Response properties of trigeminal ganglion (NV) neurons in 2-, 3-, and 4-week-old and adult rats were examined using extracellular single-unit recordings and controlled whisker stimuli. We found that the receptive field size of NV neurons is mature in 2-week-old animals while response latencies, magnitudes, and angular tuning continue to develop between 2 weeks of age and adulthood. At the earliest time recorded, NV neurons respond to stimulation of only one whisker and can be characterized as slowly or rapidly adapting (SA, RA). The proportion of SA and RA neurons remains constant during development. Consistent with known on-going myelination of NV axons, response latencies decrease with age, becoming adult-like during the third and fourth postnatal weeks for RA and SA neurons, respectively. Unexpectedly, we found that evoked response magnitudes increase several-fold during development becoming adult-like only during the fourth postnatal week. In addition, RA neurons become less selective for whisker deflection angle with age. Maturation of response magnitude and angular tuning is consistent with developmental changes in the mechanical properties of the whisker, the whisker follicle, and the surrounding tissues. The findings indicate that whisker-derived tactile inputs mature during the first postnatal month when whisker-related cortical circuits are susceptible to long-term modification by sensory experience. Thus normal developmental changes in sensory input may influence functional development of cortical circuits.     

Neurodegeneration in the somatosensory cortex after experimental diffuse brain injury

Disruption and consequent reorganization of central nervous system circuits following traumatic brain injury may manifest as functional deficits and behavioral morbidities. We previously reported axotomy and neuronal atrophy in the ventral basal (VB) complex of the thalamus, without gross degeneration after experimental diffuse brain injury in adult rats. Pathology in VB coincided with the development of late-onset aberrant behavioral responses to whisker stimulation, which lead to the current hypothesis that neurodegeneration after experimental diffuse brain injury includes the primary somatosensory barrel cortex (S1BF), which receives projection of VB neurons and mediates whisker somatosensation. Over 28 days after midline fluid percussion brain injury, argyrophilic reaction product within superficial layers and layer IV barrels at 1 day progresses into the cortex to subcortical white matter by 7 days, and selective inter-barrel septa and subcortical white matter labeling at 28 days. Cellular consequences were determined by stereological estimates of neuronal nuclear volumes and number. In all cortical layers, neuronal nuclear volumes significantly atrophied by 42–49% at 7 days compared to sham, which marginally attenuated by 28 days. Concomitantly, the number of healthy neurons was reduced by 34–45% at 7 days compared to sham, returning to control levels by 28 days. Progressive neurodegeneration, including argyrophilic reaction product and neuronal nuclear atrophy, indicates injury-induced damage and reorganization of the reciprocal thalamocortical projections that mediate whisker somatosensation. The rodent whisker barrel circuit may serve as a discrete model to evaluate the causes and consequences of circuit reorganization after diffuse brain injury.     

Tracking whisker and head movements in unrestrained behaving rodents

Due to recent advances that enable real-time electrophysiological recordings in brains of awake behaving rodents, effective methods for analyzing the large amount of behavioral data thus generated, at millisecond resolution, are required. We describe a semiautomated, efficient method for accurate tracking of head and mystacial vibrissae (whisker) movements in freely moving rodents using high-speed video. By tracking the entire length of individual whiskers, we show how both location and shape of whiskers are relevant when describing the kinematics of whisker movements and whisker interactions with objects during a whisker-dependent task and exploratory behavior.     

Responses of barrel cortex neurons in awake rats and effects of urethane anesthesia

Summary A barrel is an interconnected network of layer IV neurons that is an important component of a functional cortical column in the whisker area of the rodent primary somatosensory cortex. The present study was undertaken in order to resolve apparently conflicting findings from single-unit studies of barrel neurons conducted in rats maintained under different anesthetic conditions. Multiunit responses to controlled deflections of mystacial vibrissae were recorded from the whisker/ barrel cortex of awake, undrugged rats, and responses at the same recording site were reexamined after the animal was anesthetized with urethane. In contrast to the awake condition, stimulus-evoked responses under urethane were characterized by a large late component. Such effects were more pronounced for deflections of noncolumnar or adjacent whiskers than for the the columnar whisker. Latencies to peak responses were virtually identical for the columnar whisker in awake and urethane states (11.9 vs 11.8 ms) but were considerably longer for adjacent whisker deflections in urethane-anesthetized animals (15.5 vs 29.0 ms). The magnitudes of adjacent whisker responses, relative to the response evoked by the columnar whisker, varied with the laminar location of the recording site in awake but not in urethane-anesthetized animals; in awake rats, receptive fields were clearly smallest in the layer IV barrels. Results in the awake condition confirm those of previous studies conducted in unanesthetized or lightly sedated animals, and data obtained with urethane are comparable to others' results in urethane-anesthetized rats. The former have important implications for how barrel cortex processes information in behaving animals.     

Unsupervised whisker tracking in unrestrained behaving animals

Understanding how whisker-based tactile information is represented in the nervous system requires quantification of sensory input and observation of neural activity during whisking and whisker touch. Chronic electrophysiological methods have long been available to study neural responses in awake and behaving animals; however, methods to quantify the sensory input on whiskers have not yet been developed. Here we describe an unsupervised algorithm to track whisker movements in high-speed video recordings and to quantify the statistics of the tactile information on whiskers in freely behaving animals during haptic object exploration. The algorithm does not require human identification of whiskers, nor does it assume the shape, location, orientation, length of whiskers, or direction of the whisker movements. The algorithm performs well on temporary loss of whisker visibility and under low-light/low-contrast conditions even with inherent anisotropic noise and non-Gaussian variability in the signal. Using this algorithm, we define the speed [protraction (P), 1,081 +/- 322; retraction (R), 1,564 +/- 549 degrees /s], duration (P, 34 +/- 10; R, 24 +/- 8 ms), amplitude (P = R, 40 +/- 13 degrees ), and frequency (19 +/- 7 Hz) of active whisking in freely behaving mice. We furthermore quantify whisker deflection induced changes in whisking kinematics and calculate the statistics (i.e., speed, amplitude and duration) of whisker touch and finally show that whisker deprivation does not alter whisking kinematics during haptic exploration.     

Frequency adaptation modulates spatial integration of sensory responses in the rat whisker system

The generation of perceptual experiences requires the integration of complex spatiotemporal patterns of sensory input. The rodent whisker system is a useful model for understanding the cellular mechanisms of sensory integration, which often include the operation of local circuits distributed throughout the brain. An example is cross-whisker suppression, where the neuronal response to whisker deflection is strongly reduced by preceding deflection of a neighboring whisker. Suppressive interactions between whisker-evoked responses have largely been studied using pairs of single whisker deflections. However, rats typically sweep their whiskers across surfaces at frequencies ranging from 5 to 25 Hz. Repetitive afferent activation induces frequency-dependent adaptation of neuronal responses and alters the synaptic dynamics of circuits that play a role in suppression, suggesting that adaptation could modulate the spatial integration of whisker evoked responses. We tested this hypothesis by comparing the cross-whisker suppression of principal whisker (PW)-evoked cortical and thalamic responses when preceded by either a single deflection of an adjacent whisker (AW) or a train of AW deflections at frequencies covering the normal whisking range. We found that periodic deflection of the preceding AW significantly reduced the magnitude of cross-whisker suppression. Surprisingly, although higher frequencies resulted in greater adaptation of the AW-evoked response, the effect on suppression was independent of frequency. We suggest that these results follow from known local circuit operations at multiple levels within the afferent path. Our findings support the view that repetitive whisking subserves a transformation of the integrative and functional properties of the whisker system.     

The barrel cortex—integrating molecular, cellular and systems physiology

A challenge for neurobiology is to integrate information across many levels of research, ranging from behaviour and neuronal networks to cells and molecules. The rodent whisker signalling pathway to the primary somatosensory neocortex with its remarkable somatotopic barrel map is emerging as a key system for such integrative studies.     

Relationship between physiological response type (RA and SA) and vibrissal receptive field of neurons within the rat trigeminal ganglion

Cells within the trigeminal ganglion (Vg) encode all the information necessary for the rat to differentiate tactile stimuli, yet it is the least-studied component in the rodent trigeminal somatosensory system. For example, extensive anatomical and electrophysiological investigations have shown clear somatotopic organization in the higher levels of this system, including VPM thalamus and SI cortex, yet whether this conserved schemata exists in the Vg is unknown. Moreover although there is recent interest in recording from vibrissae-responsive cells in the Vg, it is surprising to note that the locations of these cells have not even been clearly demarcated. To address this, we recorded extracellularly from 350 sensory-responsive Vg neurons in 35 Long-Evans rats. First, we determined three-dimensional locations of these cells and found a finer detail of somatotopy than previously reported. Cells innervating dorsal facial features, even within the whisker region, were more dorsal than midline and ventral features. We also show more cells with caudal than rostral whisker receptive fields (RF), similar to that found in VPM and SI. Next, for each vibrissal cell we determined its response type classified as either rapidly (RA) or slowly (SA) adapting. We examined the relationship between vibrissal RF and response type and demonstrate similar proportions of RA and SA cells responding to any whisker. These results suggest that if RA and SA cells encode distinct features of stimuli, as previously suggested, then at the basic physiological level each whisker has similar abilities to encode for such features.     

A microcomputer program for fitting enzyme inhibition rate equations

This paper presents a computer program written in BASIC for fitting different enzyme inhibition kinetic models. The program, based on a nonlinear least-squares regression, can be run on any microcomputer with the CP/M operating system. Weighting of observed initial velocities is decided by the user by assessing constant variance, proportional variance or by incorporation of the variances calculated by a subroutine. The program also uses robust regression by bisquare weighting. All questions concerning data input, type of rate function, type of weight and the use of bisquare regression appear on the video display unit.     

ETHOWATCHER: validation of a tool for behavioral and video-tracking analysis in laboratory animals

We present a software (ETHOWATCHER^®) developed to support ethography, object tracking and extraction of kinematic variables from digital video files of laboratory animals. The tracking module allows controlled segmentation of the target from the background, extracting image attributes used to calculate the distance traveled, orientation, length, area and a path graph of the experimental animal. The ethography module allows recording of catalog-based behaviors from environment or from video files continuously or frame-by-frame. The output reports duration, frequency and latency of each behavior and the sequence of events in a time-segmented format, set by the user. Validation tests were conducted on kinematic measurements and on the detection of known behavioral effects of drugs. This software is freely available at www.ethowatcher.ufsc.br.     

An observer study comparing spot imaging regions selected by radiologists and a computer for an automated stereo spot mammography technique

We are developing an automated stereo spot mammography technique for improved imaging of suspicious dense regions within digital mammograms. The technique entails the acquisition of a full-field digital mammogram, automated detection of a suspicious dense region within that mammogram by a computer aided detection (CAD) program, and acquisition of a stereo pair of images with automated collimation to the suspicious region. The latter stereo spot image is obtained within seconds of the original full-field mammogram, without releasing the compression paddle. The spot image is viewed on a stereo video display. A critical element of this technique is the automated detection of suspicious regions for spot imaging. We performed an observer study to compare the suspicious regions selected by radiologists with those selected by a CAD program developed at the University of Michigan. True regions of interest (TROIs) were separately determined by one of the radiologists who reviewed the original mammograms, biopsy images, and histology results. We compared the radiologist and computer-selected regions of interest (ROIs) to the TROIs. Both the radiologists and the computer were allowed to select up to 3 regions in each of 200 images (mixture of 100 CC and 100 MLO views). We computed overlap indices (the overlap index is defined as the ratio of the area of intersection to the area of interest) to quantify the agreement between the selected regions in each image. The averages of the largest overlap indices per image for the 5 radiologist-to-computer comparisons were directly related to the average number of regions per image traced by the radiologists (about 50% for 1 region/image, 84% for 2 regions/image and 96% for 3 regions/image). The average of the overlap indices with all of the TROIs was 73% for CAD and 76.8% +/- 10.0% for the radiologists. This study indicates that the CAD determined ROIs could potentially be useful for a screening technique that includes stereo spot mammography imaging.     

Cellular mechanisms of motor control in the vibrissal system

In this article we discuss the experimental advantages that the vibrissal motor system offers for analysis of motor control and the specializations of this system related to the unique characteristics of whisker movements. Whisker movements are often rhythmic, fast, and bilateral. Movements of individual whiskers have simple characteristics, whereas, movements of the entire vibrissae array are complex and sophisticated. In the last few years, powerful methods for high precision tracking of whisker movements have become available. The whisker musculature is arranged to permit forward movements of individual whiskers and consists—depending on the species—mainly or exclusively of fast contracting, fast fatigable muscle fibers. Whisker motor neurons are located in the lateral facial nucleus and their cellular properties might contribute to the rhythmicity of whisking. Numerous structures provide input to the lateral facial nucleus, the most mysterious and important one being the putative central pattern generator (CPG). Although recent studies identified candidate structures for the CPG, the precise identity and the functional organization of this structure remains uncertain. The vibrissa motor cortex (VMC) is the largest motor representation in the rodent brain, and recent work has clarified its localization, subdivisions, cytoarchitectonics, and connectivity. Single-cell stimulation experiments in VMC allow determining the cellular basis of cortical motor control with unprecedented precision. The functional significance of whisker movements remains to be determined.     

Thalamocortical conduction times and stimulus-evoked responses in the rat whisker-to-barrel system

Studies of the rodent whisker system indicate that somatosensory cortical circuitry operates at a millisecond timescale to transform sensory afferent signals from the thalamus. We measured axon conduction times and whisker-evoked responses of 48 thalamocortical (TC) neurons in the rat whisker-to-barrel pathway. Conduction times were derived from spike-triggered averages of local field potentials evoked in layer 4 cortical whisker-related barrels by the spontaneous firing of individual topographically aligned neurons in the ventral posterior medial thalamus. Conduction times varied fourfold, from 0.31 to 1.34 ms, and faster conducting TC neurons responded earlier and more robustly to controlled whisker deflections. Early arrival of highly responsive TC inputs, thought to contact inhibitory barrel neurons preferentially, could prime the cortical network, rendering it more selective for later-arriving signals.     

Enhancement of cortical plasticity by behavioral training in acetylcholine-depleted adult rats

Trimming all whiskers except two on one side of an adult rat's face results in cortical plasticity in which the spared whiskers, D2 and one D-row surround whisker (either D1 or D3), evoked responses containing more spikes than the response evoked by the cut whisker (called whisker pairing plasticity). Previously we have reported that acetylcholine (ACh) depletion in cortex prevents surround D-row whisker plasticity from developing within the barrel cortex. In this study we examined whether the animal's active use of its two intact whiskers can restore some aspects of plasticity in the ACh-depleted cortex. To achieve this goal, ACh was depleted from barrel field cortex, and 14 days after the depletion surgery, whiskers were trimmed and animals were trained on a whisker-dependent gap crossing task. After 7 days of training, animals were anesthetized with urethan and prepared for single-unit recording. Training the ACh-depleted, whisker-paired animals resulted in a significant enhancement of responses to paired surround whiskers: the D-paired whisker-evoked response contained more spikes than the D-cut evoked response. We conclude that training whisker paired rats has a positive impact on response properties of neurons in S1 cortex, even in ACh-depleted animals.     

The think aloud method: a guide to user interface design

OBJECTIVES: Clinical use of computer systems has been hampered by their poorly designed user interfaces. The objective of this study was to design a user interface for a pediatric oncologists' computerized patient record with great consideration of their working behavior and of human computer interfacing principles so as to contribute to oncologists' efficiency and satisfaction in interaction with the system. METHODS: The think aloud method was used in combination with video recording to get a deep understanding of the way in which four pediatric oncologists searched through the paper-based patient record in preparing a patient visit. Protocol and video analyses was used to develop a cognitive task model reflecting pediatric oncologists' task behavior. This model was input for a prototype user interface, which was subsequently evaluated by eight other pediatric oncologists. RESULTS: The resulting computerized medical record system proved to meet pediatric oncologists' information needs and task behavior patterns. The design of the user interface minimized pediatric oncologists' work load and was highly efficient in supporting the pediatric oncologists in preparing their patient visits. The pediatric oncologists were very much satisfied with the computer system. CONCLUSIONS: It is argued that early involvement of cognitive engineering methods in the system design process may be of great help in designing systems that fully support health care professionals' work practices. The think aloud method, if applied under prescribed conditions, is a valuable information source of human task-behavior and as such a useful technique for requirements analysis in designing clinical computer systems.     

Varying responses to the rat forced-swim test under diurnal and nocturnal conditions

The paradox that experiments in behavioural pharmacology employing nocturnal rodent species are carried out almost exclusively in the resting phase of the animals' circadian cycle has remained largely unexamined and unquestioned. This is despite the fact that all major physiological systems in the body are intrinsically aligned with its natural circadian rhythm. The forced-swim test (FST) is a rodent model that is used extensively as a screening test for antidepressant activity. The objectives of the present study were to examine the behaviour of rats in the FST under diurnal and nocturnal conditions and, in addition, to profile the response of neurochemical, neuroendocrine, and cellular indices of stress at time points up to 120 min following exposure to the FST. The time spent in escape-oriented activity was significantly less when animals were tested in the dark phase. The profile of serum corticosterone and adrenal ascorbic acid concentrations indicates that the animals were less stressed by the test situation during the active (i.e., dark) phase of their circadian cycle. Similarly, increases in blood enzymatic markers of stress-induced cellular damage were less marked following FST exposure in the nocturnal period. Characteristic stress-induced increases in 5-HT turnover in the frontal cortex and amygdala observed in the diurnal phase were reversed in the nocturnal period. In conclusion, circadian differences in behaviour in the FST may be related to parallel alterations in the ability of animals to adapt to exposure to stress.