Interactions of Whisking and Touch Signals in the Rat Brainstem.

Perception is an active process, requiring the integration of both proprioceptive and exteroceptive information. In the rat's vibrissal system, a classical model for active sensing, the relative contribution of the two information streams was previously studied at the peripheral, thalamic, and cortical levels. Contributions of brainstem neurons were only indirectly inferred for some trigeminal nuclei according to their thalamic projections. The current work addressed this knowledge gap by performing the first comparative study of the encoding of proprioceptive whisking and exteroceptive touch signals in the oralis (SpVo), interpolaris (SpVi), and paratrigeminal (Pa5) brainstem nuclei. We used artificial whisking in anesthetized male rats, which allows a systematic analysis of the relative contribution of the proprioceptive and exteroceptive information streams along the ascending pathways in the absence of motor or cognitive top-down modulations. We found that (1) neurons in the rostral and caudal parts of the SpVi convey whisking and touch information, respectively, as predicted by their thalamic projections; (2) neurons in the SpVo encode both whisking and (primarily) touch information; and (3) neurons of the Pa5 encode a complex combination of whisking and touch information. In particular, the Pa5 contains a relatively large fraction of neurons that are inhibited by active touch, a response observed so far only in the thalamus. Overall, our systematic characterization of afferent responses to active touch in the trigeminal brainstem approves the hypothesized functions of SpVi neurons and presents evidence that SpVo and Pa5 neurons are involved in the processing of active vibrissal touch.SIGNIFICANCE STATEMENT The present work constitutes the first comparative study of the encoding of proprioceptive (whisking) and exteroceptive (touch) information in the rat's brainstem trigeminal nuclei, the first stage of vibrissal processing in the CNS. It shows that (1) as expected, the rostral and caudal interpolaris neurons convey primarily whisking and touch information, respectively; (2) the oralis nucleus, whose function was previously unknown, encodes both whisking and (primarily) touch touch information; (3) a subtractive computation, reported at the thalamic level, already occurs at the brainstem level; and (4) a novel afferent pathway probably ascends via the paratrigeminal nucleus, encoding both proprioceptive and exteroceptive information.

Interaction between muscles and fascia in the mystacial pad of whisking rodents.

In whisking rodents, the mystacial pad is supplied with vibrissae and contains a collagenous skeleton that is a part of the snout fascia. The collagenous skeleton is composed of three interconnected layers: superficial, deep spongy mesh and subcapsular fibrous mat. We found that the first two layers contain diverse fascial structures, such as sheets of subcutaneous connective tissue, tendons, ligaments and follicular capsules which transmit muscle efforts to vibrissae and are thus involved in whisking. Subcapsular fibrous mat is built of oriented rostro-caudal wavy fibrils. It maintains spatial arrangement of whisker follicles, provides a quick response to deformation and connects entire mystacial pad to the skull. To move vibrissae, the forces of intrinsic muscles are applied directly to the capsules of the vibrissa follicles, whereas the forces of extrinsic muscles are applied to other parts of the collagenous skeleton, which transmit the forces to the capsules. According to the spatial distribution and anchoring sites of the muscles and fascia, extrinsic muscles provide vibrissa protraction or retraction by pulling the superficial layer of the collagenous skeleton rostral or caudal, respectively. Vibrissae can be also retracted when the efforts of extrinsic muscles are applied to the subcapsular fibrous mat. When the muscles relax, fascial structures return the vibrissae to their resting position. The deep spongy layer encompasses vibrissal follicles providing a uniform distribution of stresses and strains during whisking. In the mystacial pad, fascia is a dominant type of tissue that maintains the integrity of the vibrissa motor plant, translates muscular momentum to the vibrissae, and plays a role in vibrissae movements.

Muscular Basis of Whisker Torsion in Mice and Rats.

Whisking mammals move their whiskers in the rostrocaudal and dorsoventral directions with simultaneous rolling about their long axes (torsion). Whereas muscular control of the first two types of whisker movement was already established, the anatomic muscular substrate of the whisker torsion remains unclear. Specifically, it was not clear whether torsion is induced by asymmetrical operation of known muscles or by other largely unknown muscles. Here, we report that mystacial pads of newborn and adult rats and mice contain oblique intrinsic muscles (OMs) that connect diagonally adjacent vibrissa follicles. Each of the OMs is supplied by a cluster of motor end plates. In rows A and B, OMs connect the ventral part of the rostral follicle with the dorsal part of the caudal follicle. In rows C-E, in contrast, OMs connect the dorsal part of the rostral follicle to the ventral part of the caudal follicle. This inverse architecture is consistent with previous behavioral observations [Knutsen et al.: Neuron 59 (2008) 35-42]. In newborn mice, torsion occurred in irregular single twitches. In adult anesthetized rats, microelectrode mediated electrical stimulation of an individual OM that is coupled with two adjacent whiskers was sufficient to induce a unidirectional torsion of both whiskers. Torsional movement was associated with protracting movement, indicating that in the vibrissal system, like in the ocular system, torsional movement is mechanically coupled to horizontal and vertical movements. This study shows that torsional whisker rotation is mediated by specific OMs whose morphology and attachment sites determine rotation direction and mechanical coupling, and motor innervation determines rotation dynamics. Anat Rec, 300:1643-1653, 2017. © 2017 Wiley Periodicals, Inc.

On-going computation of whisking phase by mechanoreceptors.

To attribute spatial meaning to sensory information, the state of the sensory organ must be represented in the nervous system. In the rodent's vibrissal system, the whisking-cycle phase has been identified as a key coordinate, and phase-based representation of touch has been reported in the somatosensory cortex. Where and how phase is extracted in the ascending afferent pathways remains unknown. Using a closed-loop interface in anesthetized rats, we found that whisking phase is already encoded in a frequency- and amplitude-invariant manner by primary vibrissal afferents. We found that, for naturally constrained whisking dynamics, such invariant phase coding could be obtained by tuning each receptor to a restricted kinematic subspace. Invariant phase coding was preserved in the brainstem, where paralemniscal neurons filtered out the slowly evolving offset, whereas lemniscal neurons preserved it. These results demonstrate accurate, perceptually relevant, mechanically based processing at the sensor level.

Structure-function correlations of rat trigeminal primary neurons: Emphasis on club-like endings, a vibrissal mechanoreceptor.

This study focuses on the structure and function of the primary sensory neurons that innervate vibrissal follicles in the rat. Both the peripheral and central terminations, as well as their firing properties were identified using intracellular labelling and recording in trigeminal ganglia in vivo. Fifty-one labelled neurons terminating peripherally, as club-like, Merkel, lanceolate, reticular or spiny endings were identified by their morphology. All neurons responded robustly to air puff stimulation applied to the vibrissal skin. Neurons with club-like endings responded with the highest firing rates; their peripheral processes rarely branched between the cell body and their terminal tips. The central branches of these neurons displayed abundant collaterals terminating within all trigeminal nuclei. Analyses of three-dimensional reconstructions reveal a palisade arrangement of club-like endings bound to the ringwulst by collagen fibers. Our morphological findings suggest that neurons with club-like endings sense mechanical aspects related to the movement of the ringwulst and convey this information to all trigeminal nuclei in the brainstem.

Pre-neuronal morphological processing of object location by individual whiskers.

In the vibrissal system, touch information is conveyed by a receptorless whisker hair to follicle mechanoreceptors, which then provide input to the brain. We examined whether any processing, that is, meaningful transformation, occurs in the whisker itself. Using high-speed videography and tracking the movements of whiskers in anesthetized and behaving rats, we found that whisker-related morphological phase planes, based on angular and curvature variables, can represent the coordinates of object position after contact in a reliable manner, consistent with theoretical predictions. By tracking exposed follicles, we found that the follicle-whisker junction is rigid, which enables direct readout of whisker morphological coding by mechanoreceptors. Finally, we found that our behaving rats pushed their whiskers against objects during localization in a way that induced meaningful morphological coding and, in parallel, improved their localization performance, which suggests a role for pre-neuronal morphological computation in active vibrissal touch.

Temporal and spatial characteristics of vibrissa responses to motor commands.

A mechanistic description of the generation of whisker movements is essential for understanding the control of whisking and vibrissal active touch. We explore how facial-motoneuron spikes are translated, via an intrinsic muscle, to whisker movements. This is achieved by constructing, simulating, and analyzing a computational, biomechanical model of the motor plant, and by measuring spiking to movement transformations at small and large angles using high-precision whisker tracking in vivo. Our measurements revealed a supralinear summation of whisker protraction angles in response to consecutive motoneuron spikes with moderate interspike intervals (5 ms < Deltat < 30 ms). This behavior is explained by a nonlinear transformation from intracellular changes in Ca(2+) concentration to muscle force. Our model predicts the following spatial constraints: (1) Contraction of a single intrinsic muscle results in movement of its two attached whiskers with different amplitudes; the relative amplitudes depend on the resting angles and on the attachment location of the intrinsic muscle on the anterior whisker. Counterintuitively, for a certain range of resting angles, activation of a single intrinsic muscle can lead to a retraction of one of its two attached whiskers. (2) When a whisker is pulled by its two adjacent muscles with similar forces, the protraction amplitude depends only weakly on the resting angle. (3) Contractions of two adjacent muscles sums up linearly for small amplitudes and supralinearly for larger amplitudes. The model provides a direct translation from motoneuron spikes to whisker movements and can serve as a building block in closed-loop motor-sensory models of active touch.

Layer-specific touch-dependent facilitation and depression in the somatosensory cortex during active whisking.

Brains adapt to new situations by retuning their neurons. The most common form of neuronal adaptation, typically observed with repetitive stimulations of passive sensory organs, is depression (responses gradually decrease until stabilized). We studied cortical adaptation when stimuli are acquired by active movements of the sensory organ. In anesthetized rats, artificial whisking was induced at 5 Hz, and activity of individual neurons in layers 2-5 was recorded during whisking in air (Whisking condition) and whisking against an object (Touch condition). Response strengths were assessed by spike counts. Input-layer responses (layers 4 and 5a) usually facilitated during the whisking train, whereas superficial responses (layer 2/3) usually depressed. In layers 2/3 and 4, but not 5a, responses were usually stronger during touch trials than during whisking in air. Facilitations were specific to the protraction phase; during retraction, responses depressed in all layers and conditions. These dynamic processes were accompanied by a slow positive wave of activity progressing from superficial to deeper layers and lasting for approximately 1 s, during the transient phase of response. Our results indicate that, in the cortex, adaptation does not depend only on the level of activity or the frequency of its repetition but rather on the nature of the sensory information that is conveyed by that activity and on the processing layer. The input and laminar specificities observed here are consistent with the hypothesis that the paralemniscal layer 5a is involved in the processing of whisker motion, whereas the lemniscal barrels in layer 4 are involved in the processing of object identity.

Active construction of percepts about object location.

Mammals acquire much of their sensory information by actively moving their sensory organs. Rats, in particular, scan their surrounding environment with their whiskers. This form of active sensing induces specific patterns of temporal encoding of sensory information, which are based on a conversion of space into time via sensor movement. We investigate the ways in which object location is encoded by the whiskers and decoded by the brain. We recorded from first-order neurons located in the trigeminal ganglion (TG) of anaesthetized rats during epochs of artificial whisking induced by electrical stimulation of the facial motor nerve. We found that TG neurons encode the three positional coordinates with different codes. The horizontal coordinate (along the backward-forward axis) is encoded by two encoding schemes, both relying on the firing times of one type of TG neuron, the 'contact cell'. The radial coordinate (from face outward) is encoded primarily by the firing magnitude of another type of TG neurons, the 'pressure cell'. The vertical coordinate (from ground up) is encoded by the identity of activated neurons. The decoding schemes of at least some of these sensory cues, our data suggest, are also active: cortical representations are generated by a thalamic comparison of cortical expectations with incoming sensory data.

Responses of trigeminal ganglion neurons to the radial distance of contact during active vibrissal touch.

Rats explore their environment by actively moving their whiskers. Recently, we described how object location in the horizontal (front-back) axis is encoded by first-order neurons in the trigeminal ganglion (TG) by spike timing. Here we show how TG neurons encode object location along the radial coordinate, i.e., from the snout outward. Using extracellular recordings from urethane-anesthetized rats and electrically induced whisking, we found that TG neurons encode radial distance primarily by the number of spikes fired. When an object was positioned closer to the whisker root, all touch-selective neurons recorded fired more spikes. Some of these cells responded exclusively to objects located near the base of whiskers, signaling proximal touch by an identity (labeled-line) code. A number of tonic touch-selective neurons also decreased delays from touch to the first spike and decreased interspike intervals for closer object positions. Information theory analysis revealed that near-certainty discrimination between two objects separated by 30% of the length of whiskers was possible for some single cells. However, encoding reliability was usually lower as a result of large trial-by-trial response variability. Our current findings, together with the identity coding suggested by anatomy for the vertical dimension and the temporal coding of the horizontal dimension, suggest that object location is encoded by separate neuronal variables along the three spatial dimensions: temporal for the horizontal, spatial for the vertical, and spike rate for the radial dimension.

Encoding of vibrissal active touch.

Mammals acquire much of their sensory information by actively moving their sensory organs. Yet, the principles of encoding by active sensing are not known. Here we investigated the encoding principles of active touch by rat whiskers (vibrissae). We induced artificial whisking in anesthetized rats and recorded from first-order neurons in the trigeminal ganglion. During active touch, first-order trigeminal neurons presented a rich repertoire of responses, which could not be inferred from their responses to passive deflection stimuli. Individual neurons encoded four specific events: whisking, contact with object, pressure against object, and detachment from object. Whisking-responsive neurons fired at specific deflection angles, reporting the actual whiskers' position with high precision. Touch-responsive neurons encoded the horizontal coordinate of objects' position by spike timing. These findings suggest two specific encoding-decoding schemes for horizontal object position in the vibrissal system.