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J C Houk

Publications and source records attributed to J C Houk.

At least 19 recordsLinked to original sources

Evidence for GABAergic interneurons in the red nucleus of the painted turtle.

Immunocytochemical and electrophysiological evidence supporting the presence of GABAergic interneurons in the turtle red nucleus is presented. Injections of HRP into the spinal cord produced labeling of large neurons in the contralateral red nucleus. The peroxidase-antiperoxidase (PAP) method revealed smaller cells immunoreactive to an antibody against glutamate decarboxylase (GAD), the synthetic enzyme for the inhibitory neurotransmitter GABA, that were interspersed among larger immunonegative neurons. Similar small neurons were densely immunostained by antibodies to GABA-glutaraldehyde conjugates obtained from different sources and applied according to pre-embedding and postembedding protocols. Rubrospinal neurons retrogradely labeled with HRP measured 16 and 27 microns in mean minor and major cell body diameters, while GABA-like immunopositive neurons situated within the red nucleus measured 7 and 13 microns. There was very little overlap in soma size between the two cell populations. Therefore, we suggest that the GAD- and GABA-positive neurons may be local inhibitory interneurons. This notion is further supported by observations of pre-embedding immunostaining for GAD and postembedding immunostaining for GABA showing that the turtle red nucleus is amply innervated by immunoreactive axon terminals. These puncta are closely apposed to cell bodies and dendrites of both immunonegative large neurons and immunopositive small neurons. Moreover, immunogold staining at the electron microscopic level demonstrated that GABA-like immunoreactive axon terminals with pleomorphic synaptic vesicles formed symmetric synapses with cell bodies and dendrites of the two types of red nucleus cells. These ultrastructural features are commonly assumed to indicate inhibitory synapses. A moderately labeled bouton with round vesicles and asymmetric synapses was also observed. In addition, the two types of red nucleus neurons received asymmetric axosomatic and axodendritic synapses with GABA-negative boutons provided with round vesicles, features usually associated with excitatory functions. To obtain electrophysiological evidence for inhibition, intracellular recordings from red nucleus neurons were conducted using an in vitro brainstem-cerebellum preparation from the turtle. Small, spontaneous IPSPs were recorded from 7 out of 14 red nucleus cells studied. These morphological and physiological results provide strong support for concluding that the turtle red nucleus, like its mammalian counterpart, contains GABAergic inhibitory interneurons. While we have not identified the main source of input to these interneurons, in view of the scarce development of the reptilian cerebral cortex, this input is unlikely to come from the motor cortex as it does in mammals.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

Sulforhodamine labeling of neural circuits engaged in motor pattern generation in the in vitro turtle brainstem-cerebellum.

A fluorescent molecular probe was used in combination with a novel in vitro preparation to study spatial patterns of neural activity associated with motor pattern generation. The in vitro brainstem-cerebellum preparation takes advantage of the turtle's unusual resistance to anoxia to preserve the entire neural network that connects the cerebellum, red nucleus, and reticular formation. This preparation was bathed in a 0.01% solution of sulforhodamine while it was activated unilaterally by electrical stimulation of the dorsal quadrant of the spinal cord for 1 hr. Sulforhodamine is a small, sulfonated, highly charged fluorescent molecule that is taken up by endocytosis. To examine its distribution in the cerebellum and brainstem, coronal sections were prepared and viewed under epifluorescence illumination. Distinctive spatial patterns of labeling were associated with unilateral electrical stimulation of the in vitro network, suggesting that dye uptake was activity dependent. Blockade of uptake with altered magnesium and calcium concentrations indicated that single spike discharge evoked ortho- or antidromically was insufficient to induce dye uptake. Instead, sulforhodamine staining correlated with the presence of burst discharge that was recorded extracellularly from the red nucleus. Blockade of burst discharge with excitatory amino acid receptor antagonists prevented dye uptake in the red nucleus, the lateral cerebellar nucleus, and other structures that are known to be interconnected by recurrent anatomical pathways. These results suggest that sulforhodamine is internalized by intensely active neurons. The spatial distributions of label support the hypothesis that burst discharges in the turtle red nucleus are mediated by excitatory amino acid neurotransmitters and sustained by recurrent excitation in cerebellorubral synaptic pathways. Positive feedback in these recurrent pathways may provide an important driving force for the generation of motor programs that control limb movements.

Animals

Red nucleus: role in motor control.

Experimental reports in the past year have provided a better understanding of the motor functions of excitatory and inhibitory neurotransmitters in the red nucleus, and of the sensorimotor properties of single rubral neurons. These data fit well within the framework of a neural network model of the rubrocerebellar system.

Animals

Role of excitatory amino acids in mediating burst discharge of red nucleus neurons in the in vitro turtle brain stem-cerebellum.

1. Bursts of discharge have been recorded in the red nucleus in several species and are thought to represent the expression of motor commands. A cerebellorubral circuit comprised of recurrent connections among the cerebellum, red nucleus, and reticular formation was postulated to function as a positive feedback loop that generates these motor commands and transmits them to the spinal cord via the rubrospinal pathway. We have used an in vitro preparation from the turtle that leaves the circuitry connecting the cerebellum, brain stem, and spinal cord intact to study the role of excitatory amino acid neurotransmitters and recurrent excitation in mediating the generation of burst discharges in the red nucleus. 2. Burst discharges were recorded extracellularly from single cells in the red nucleus in response to single pulse or brief train stimulation of the contralateral spinal cord or brief train stimuli applied to the ipsilateral cerebellar cortex. The firing characteristics and pharmacologic sensitivities of the bursts were independent of the type of stimulus used. The bursts had long durations ranging from 2 to 17 s and showed spike frequency adaptation. 3. Transection of the cerebellar peduncle, which eliminates inhibition impinging onto the cerebellorubral circuit, greatly enhanced the spontaneous activity and burst discharges recorded in the contralateral red nucleus. Furthermore, bath application of a solution containing elevated levels of calcium and magnesium blocked the expression of burst discharges even though synaptic activation of the neurons was not blocked. 4. The possibility that excitatory amino acid receptors mediate burst responses in the red nucleus was investigated in light of the antagonistic effects of elevated magnesium ions on bursting. Bath application of 100 microns DL-2-amino-5-phosphonovaleric acid (APV), a specific N-methyl-D-aspartate (NMDA) receptor antagonist; [10 microM 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX)], a specific non-NMDA receptor antagonist; or 100 microM, DL-2-amino-4-phosphonobutyric acid (AP4), an agonist of a fourth class of excitatory amino acid receptor, blocked burst activity in the red nucleus. With a multibarreled pipette for simultaneous ejection of drug and recording, iontophoresis of APV or CNQX into the red nucleus blocked bursting whereas AP4 failed to show a significant effect. These data suggest that red nucleus neurons have both NMDA and non-NMDA receptors. The site of action of the AP4-sensitive receptor appears to be elsewhere in the cerebellorubral circuit. 5. Iontophoretic application of excitatory amino acid receptor agonists NMDA and quisqualate (Q) induced excitation of red nucleus neurons.(ABSTRACT TRUNCATED AT 400 WORDS)

2-Amino-5-phosphonovalerate

Inhibition of sensory responses of cat inferior olive neurons produced by stimulation of red nucleus.

1. The sensory responsiveness of cells in the inferior olive is known to be suppressed during certain phases of active movement. These experiments were designed to test the possibility that activity in the rubrospinal pathway contributes to this suppression. We recorded from cells sensitive to light touch located in one of the divisions of the inferior olive, the rostral dorsal accessory olive (rDAO), in cats anesthetized with pentobarbitol sodium. Responsiveness to peripheral stimuli was tested during and after trains of conditioning stimuli delivered to the rubrospinal pathway. 2. All 44 cells in our sample of rDAO neurons showed an inhibition of responsiveness to peripheral stimuli after conditioning stimulation of the rubrospinal pathway. Typical conditioning trains consisted of 0.2-ms pulses at 200 Hz for 100 ms. The mean current required for a reduction in firing probability to 0.5 was 31 microA. Slight increases in intensity often completely inhibited responses to peripheral stimuli. 3. Inhibition of responsiveness showed a delayed time course. Peak inhibition occurred approximately 50 ms after the last pulse in the conditioning train. In many cases there was no demonstrable inhibition during the conditioning train. Increases of train frequency, train duration, or stimulus intensity produced stronger and broader periods of olivary inhibition. 4. The lowest threshold points for eliciting rDAO inhibition coincided with either the magnocellular red nucleus (RNm) or the rubrospinal tract (RST). Stimulation at RST sites produced inhibition of responses in the contralateral but not in the ipsilateral rDAO. Transection of the RST in the upper brain stem blocked the inhibition produced by red-nucleus stimulation without altering the inhibition produced by tract stimulation caudal to the transection. The inhibitory effects thus appear to be caused by activation of the rubrospinal pathway. 5. The inhibitory timing observed in this study may be appropriate for explaining the suppression of olivary responsiveness to contact that has been observed in awake animals. Bursts of movement-related, red nucleus discharge often cease approximately 50 ms before the end of movement. This timing would allow peak inhibition to develop at approximately the time of contact with an object at the end of a goal-directed limb movement.

Animals

An in vitro preparation for studying motor pattern generation in the cerebellorubrospinal circuit of the turtle.

In vivo studies in mammals have suggested that the cerebellorubrospinal circuit functions as a recurrent excitatory loop that generates motor commands and transmits them to the spinal cord via the rubrospinal pathway. Here we describe an in vitro preparation from the turtle exhibiting functional synaptic connections between the cerebellum, brainstem and upper spinal cord that is suitable for detailed analysis of this circuit. Electrical stimulation of the spinal cord was used to activate the cerebellorubrospinal circuit while activity was sampled with extracellular recordings from single cells in the red nucleus. Single units responded to stimulation with short and long latency synaptic responses, in addition to antidromic activation. Some cells showed bursts of activity lasting several hundred milliseconds suggesting the presence of recurrent excitation. Interruption of Purkinje cell inhibitory input impinging on the cerebellorubrospinal loop prolonged bursting and enhanced spontaneous activity. This preparation should facilitate the examination of the role of the cerebellorubrospinal circuit in motor pattern generation.

Action Potentials

Control strategies in physiological systems.

In this paper, written for a general audience, I review and contrast various strategies that the body uses to control homeostasis and movement. Messages, signals, communication channels, and control systems are dealt with from both a cellular and an integrative perspective. The major global control strategies are feedback, feedforward, and adaptive control, and examples of each are presented to highlight advantageous and disadvantageous features. Many physiological systems use these three strategies in combination.

Animals

Selective projections from the cat red nucleus to digit motor neurons.

Classical studies of the cat rubrospinal tract describe dense terminations in spinal laminae V-VII and an absence of any significant projection to lamina IX. In contrast, our recent studies, utilizing the anterograde transport of wheat germ agglutinin conjugated with horseradish peroxidase, have demonstrated a consistent and circumscribed area of label in lamina IX at caudal cervical segments. The present study was undertaken to determine the distribution of rubrospinal terminals among motor neurons in lamina IX as well as to identify the likely target muscles of those motor neurons located near rubrospinal terminals. We injected wheat germ agglutinin-horseradish peroxidase into the red nucleus and unconjugated horseradish peroxidase into selected forearm muscles of the same side of the body. The locations of rubrospinal terminals showing anterograde label on one side of the spinal cord could then be compared with the locations of motor neurons showing retrograde label on the opposite side of the cord. The results demonstrated a clear focus of rubrospinal terminals in the lateral and dorsal portions of the ventral horn beginning at C8 and extending through rostral T1. No other segments of the spinal cord showed a focus of rubrospinal terminations in lamina IX. Retrogradely labeled motor neurons from the muscle injections showed that the rubrospinal terminals overlap extensively with motor neuronal pools supplying distal forearm muscles. Several lines of evidence indicate that the terminals are from rubrospinal fibers and are not due to transneuronal transport.

Animals

Somatotopic alignment between climbing fiber input and nuclear output of the cat intermediate cerebellum.

The rostral dorsal accessory olive (rDAO) contains a detailed somatosensory map of the entire contralateral body surface. The rDAO projects to the anterior interpositus nucleus (NIA) directly as well as indirectly by way of Purkinje cells in cerebellar cortex. NIA maintains a topographic relation to different levels of the spinal cord through a relay in the magnocellular red nucleus (RNm) and, thus, contains a motor somatotopy. By using bidirectional transport of WGA-HRP, we demonstrate that the sensory somatotopy of rDAO aligns with the motor somatotopy of NIA. It is likely that rDAO information supplied to the cerebellum from a specific part of the body is used to influence movements restricted to that same body part.

Animals

Limb specific connections of the cat magnocellular red nucleus.

Afferent and efferent connections of the limb specific divisions of the cat magnocellular red nucleus (RNm) were traced using the bidirectional transport of wheatgerm agglutinin-horseradish peroxidase complex (WGA-HRP). Injection sites within forelimb or hindlimb RNm regions were identified by microelectrode recording and confirmed by the position of labeled rubrospinal terminals. Additional injections into structures that project to, or receive input from, RNm confirmed the somatotopic organization of these pathways. The forelimb region of RNm receives input from the posteriolateral part of the anterior interpositus nucleus (NIA) and the intermediate part of the posterior interpositus nucleus (NIP). The hindlimb region of RNm receives input from anteriomedial NIA and medial NIP. Terminals of NIA cells densely fill all of RNm, but terminals of NIP cells form a half shell on the medial, ventral, and posterior borders of RNm without encroaching on RNm's lateral edge or central core. Forelimb and hindlimb RNm are reciprocally connected with the caudal cuneate and gracile nuclei respectively. There is little or no input to RNm from the medial or lateral cerebellar nuclei. Forelimb RNm, which also contains a face representation, projects to the lateral reticular nucleus, cell group f of the inferior vestibular nucleus, the facial nucleus, the main sensory nucleus of the trigeminal nerve, the caudal cuneate nucleus, the parvicellular reticular formation, and cervical segments of the spinal cord. A few fibers from forelimb RNm project directly to motor neurons in the lower cervical cord. Hindlimb RNm projects to only the lateral reticular nucleus, gracile nucleus, and lower spinal segments. Forelimb and hindlimb RNm project to different regions of the lateral reticular nucleus with some overlap.

Afferent Pathways

A model of the motor servo: incorporating nonlinear spindle receptor and muscle mechanical properties.

A model for the stretch reflex is proposed incorporating a nonlinear description of muscle receptor behavior, a delay in the reflex loop and a model of muscle mechanical properties. The model adequately describes the nonlinear response properties of EMG and force to constant ramps in loading and unloading direction. The EMG responses during the ramp and at ramp plateau could be simulated adequately for all ramp velocities except for high stretch velocities where EMG activity appeared in bursts, presumably due to spinal nonlinearities. Force responses during ramp stretches could be simulated except at ramp plateau, where the measured force response decayed slower than the simulated responses. The model also explained that EMG and force responses during ramp stretches after a displacement of about 1 cm could be approximately described by a product relationship between a position-related term and a low-fractional power of velocity. During unloading ramps the model did not predict a clear velocity dependence in agreement with the data.

Electromyography

Correlation and spectral analysis of relations between single unit discharge and muscle activities.

Correlation and spectral analysis was used to study functional linkages between single-cell discharge in the magnocellular red nucleus and the electromyographic activity of several limb muscles. Long sequences of unit discharge and EMG activity were recorded while feline subjects performed a food retrieval task. Unit discharge and muscle activity were patterned in bursts that corresponded to different phases of the task. There was sufficient variability in the parameters of these bursts to regard the signals as pseudorandom variables, thus facilitating a correlation analysis. Cross-correlation functions computed between unit discharge and each muscle EMG served to characterize the strength and reliability of linkages between a single unit and various limb muscles. Auto-correlation and auto-spectral density functions provided summary measures of the temporal and frequency characteristics of the signals. Power in the signals was concentrated in a behaviorally relevant range (0.2-8 Hz). Coherence functions showed peaks that indicated which frequency components were well correlated with unit discharge. Two-sided impulse responses served to characterize the dynamic properties of the linkages. These methods are shown to be valuable in characterizing noncausal, as well as causal, linkages through multisynaptic pathways in the nervous system.

Action Potentials

Functional and anatomic differentiation between parvicellular and magnocellular regions of red nucleus in the monkey.

Single unit recording in awake monkeys was used to search for functional differences between the two divisions of the red nucleus, and anatomical tracing of WGA-HRP was used to investigate inputs to the two divisions. We studied a total of 323 units in 4 red nuclei of two monkeys. Recording sites were identified in histological sections by the locations of lesions and the reconstruction of electrode tracks. Of the units in the RNm 98.5% discharged in high frequency bursts during movement. Only 52% showed reliable responses to somatosensory stimulation, and the responses observed were weaker than the movement-related discharge. None of the units recorded in the RNp showed strong movement-related discharge, and 51% were completely unresponsive during both motor and sensory tests. A dorsolateral group of medium-sized cells that overlaps the rostral half of the main RNm and the caudal pole of RNp appears to represent an extension of the magnocellular region. Retrograde transport of WGA-HRP indicated that some of these cells are rubrospinal neurons. Furthermore, the discharge properties of dorsolateral neurons are like the main RNm neurons, except for lower discharge rates and smaller spike amplitudes. Mouth movements are strongly represented in the dorsolateral region. Anterograde transport of WGA-HRP from the motor cortex demonstrated dense terminal label in RNp as contrasted with light label in RNm. Retrograde transport of WGA-HRP from RNm labeled many more cells in the cerebellar interpositus nucleus than in motor cortex. We concluded that input to RNm from the cerebellum is the likely source of the strong movement-related activity recorded from cells in the RNm. The absence of appreciable movement-related activity in parvicellular red nucleus provides a clear functional distinction between this division and the magnocellular division of the red nucleus.

Animals

Magnocellular red nucleus activity during different types of limb movement in the macaque monkey.

Three hundred and thirty-four neurones located in the magnocellular division of red nucleus (r.n.m.) were studied in three alert macaque monkeys. These cells had low discharge rates at rest and produced high frequency bursts during movement. Single cells were selectively active for movement of one body part, and a motor somatotopy was evident. From dorsal to ventral we encountered cells related to movements of the face, contralateral upper limb, contralateral lower limb and, in one case, the tail. Free-form tests indicated that 76% of upper-limb cells were preferentially related to hand and finger movements, and 84% of lower limb cells were preferentially related to foot and toe movements. Quantitative tests of movement relations were based on depths of modulation in discharge rate recorded while the monkeys operated several devices that served as manipulanda in a tracking task; each device elicited a different movement. We conducted 220 tests on eighty-one cells using eleven devices. The modulation in discharge rate exceeded a 50 pulses/s criterion level in seventy-nine cases; eleven were well related to proximal movements, twenty-two to movements of the digits and forty-six to a co-ordinated hand movement elicited by a device called the twister. Both unidirectional and bidirectional patterns of bursting were frequent. A few cells showed reciprocal patterns consisting of a large increase in rate for one direction of movement and a small decrease for the other. The bursts in discharge preceded movement onset (97% of 132 cases) by an average of 135 ms. Electromyographic activity in forearm muscles preceded movement by about 55 ms. In some cases we recorded from a single cell while the monkey operated two, three or four devices. Depth of modulation on the twister device was twice that on a proximal device in nine cases whereas one case showed a proximal device preference; five cases showed overlap. Comparison between twister and digits yielded ten cases of twister preference, four of digit preference and twenty cases of overlap. A directional preference was found for ten out of eleven cells responsive during active movement restricted to the metacarpo-phalangeal joints, and in all cases the preferred direction was extension. Responses to natural somatosensory stimulation were weak or absent for the majority of cells. When present, sensory fields were confined to the same limb but were frequently out of register with motor fields.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials

Relation between red nucleus discharge and movement parameters in trained macaque monkeys.

Correlation and regression analyses were performed on thirty-three of the magnocellular red nucleus cells described in the previous paper. We sought to test for reliable relations between the parameters of individual tracking movements and corresponding bursts of neural discharge. High correlations were found between the following burst and movement parameters: (i) burst latency versus movement latency; (ii) burst duration versus movement duration; (iii) burst frequency versus movement velocity and (iv) number of spikes in the burst versus movement amplitude. Cells were ranked according to the average of the duration, velocity and amplitude correlation coefficients. The top twenty cells had average correlation coefficients ranging from 0.69 to 0.88 for their preferred movement. These cases were judged most likely to reveal the control functions of the red nucleus, and the following points refer to this sample. Burst onset led movement onset by 118 +/- 23 ms, and burst offset led movement offset by 50 +/- 38 ms. Burst duration increased as the duration of the movement increased (r = 0.87 +/- 0.11). The duration of the burst was approximately equal to movement duration (slope of 0.99 +/- 0.16) plus a constant (72 +/- 34 ms) throughout a broad range. Average discharge rate during the burst increased with average movement velocity (r = 0.69 +/- 0.15). The slope of the relation was 0.36 +/- 0.21 (pulses/s)/(deg/s) of joint rotation. The regression lines had consistent upward offsets (56 +/- 15 pulses/s) that exceeded the spontaneous discharge rate (17 +/- 10 pulses/s). The number of spikes in the burst increased with movement amplitude independent of velocity (r = 0.72 +/- 0.11). The slope of the relation was 0.62 spikes/deg and the offset was 13 +/- 4 spikes. The preferred movement was co-ordinated hand in fifteen cases, digit in three, elbow in one and shoulder in one. When these cells were tested with an alternate movement, the failure rate (cases in which a burst did not accompany a movement) increased from 1.4 to 20%, and the correlation coefficients generally were low and lacked significance. Cells in the top twenty had directionally specific responses, low variance in lead time, large depths of modulation (41-118 pulses/s) and low failure rates. Cells that failed to show strong parametric correlations often had one or more of the former attributes. It appears that high parametric correlations with individual movements are particularly restrictive criteria of relatedness.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials

Inferior olivary neurons in the awake cat: detection of contact and passive body displacement.

We have recorded from 306 neurons in the inferior olive of six alert cats. Most of the cats were trained to perform a simple task with the forelimb. We observed the neural responses to a wide variety of cutaneous and proprioceptive stimuli, as well as responses during spontaneous and learned active movements. Neurons responsive to somatosensory stimulation were found in all parts of the inferior olive, and they were roughly evenly divided between those responsive to cutaneous stimulation and those responsive to proprioceptive stimulation. In the dorsal accessory olive all neurons were responsive to somatosensory stimulation. In the medial accessory nucleus 88% and in the principal olive 74% of cells were responsive to somatosensory stimulation. Cells responsive to cutaneous stimulation usually had small receptive fields, commonly on the paw. These cells had low-threshold responses to one or more forms of cutaneous stimulation and typically fired one spike at the onset of the stimulus on 80% or more of stimulus applications. Cells responsive to proprioceptive stimulation most commonly responded to passive displacements of a limb. These cells were often very sensitive, responding to linear displacements of less than 1 cm in one specific direction. No cells in our sample responded reliably during active movement by the animal. Only 21% of cells responding to passive proprioceptive stimulation showed any modulation during active movement, and the modulation was weak. Likewise, cells responsive to cutaneous stimulation generally failed to respond when a similar stimulus was produced by an active movement by the animal. Exceptions to this were stimuli produced during exploratory movements or when the receptive field unexpectedly made contact with an object during active movement. Electrical stimulation applied in the inferior olive failed to evoke movements or to modify ongoing movement. Our results are consistent with the hypothesis that inferior olivary neurons function as somatic event detectors responding particularly reliably to unexpected stimuli.

Animals