Recruitment in retractor bulbi muscle during eyeblink conditioning: EMG analysis and common-drive model.

To analyze properly the role of the cerebellum in classical conditioning of the eyeblink and nictitating membrane (NM) response, the control of conditioned response dynamics must be better understood. Previous studies have suggested that the control signal is linearly related to the CR as a result of recruitment within the accessory abducens motoneuron pool, which acts to linearize retractor bulbi muscle and NM response mechanics. Here we investigate possible recruitment mechanisms. Data came from simultaneous recordings of NM position and multiunit electromyographic (EMG) activity from the retractor bulbi muscle of rabbits during eyeblink conditioning, in which tone and periocular shock act as conditional and unconditional stimuli, respectively. Action potentials (spikes) were extracted and classified by amplitude. Firing rates of spikes with different amplitudes were analyzed with respect to NM response temporal profiles and total EMG spike firing rate. Four main regularities were revealed and quantified: 1) spike amplitude increased with response amplitude; 2) smaller spikes always appeared before larger spikes; 3) subsequent firing rates covaried for spikes of different amplitude, with smaller spikes always firing at higher rates than larger ones; and 4) firing-rate profiles were approximately Gaussian for all amplitudes. These regularities suggest that recruitment does take place in the retractor bulbi muscle during conditioned NM responses and that all motoneurons receive the same command signal (common-drive hypothesis). To test this hypothesis, a model of the motoneuron pool was constructed in which motoneurons had a range of intrinsic thresholds distributed exponentially, with threshold linearly related to EMG spike amplitude. Each neuron received the same input signal as required by the common-drive assumption. This simple model reproduced the main features of the data, suggesting that conditioned NM responses are controlled by a common-drive mechanism that enables simple commands to determine response topography in a linear fashion.

Evidence from retractor bulbi EMG for linearized motor control of conditioned nictitating membrane responses.

Classical conditioning of nictitating membrane (NM) responses in rabbits is a robust model learning system, and experimental evidence indicates that conditioned responses (CRs) are controlled by the cerebellum. It is unknown whether cerebellar control signals deal directly with the complex nonlinearities of the plant (blink-related muscles and peripheral tissues) or whether the plant is linearized to ensure a simple relation between cerebellar neuronal firing and CR profile. To study this question, the retractor bulbi muscle EMG was recorded with implanted electrodes during NM conditioning. Pooled activity in accessory abducens motoneurons was estimated from spike trains extracted from the EMG traces, and its temporal profile was found to have an approximately Gaussian shape with peak amplitude linearly related to CR amplitude. The relation between motoneuron activity and CR profiles was accurately fitted by a first-order linear filter, with each spike input producing an exponentially decaying impulse response with time constant of order 0.1 s. Application of this first-order plant model to CR data from other laboratories suggested that, in these cases also, motoneuron activity had a Gaussian profile, with time-of-peak close to unconditioned stimulus (US) onset and SD proportional to the interval between conditioned stimulus and US onsets. These results suggest that for conditioned NM responses the cerebellum is presented with a simplified "virtual" plant that is a linearized version of the underlying nonlinear biological system. Analysis of a detailed plant model suggests that one method for linearising the plant would be appropriate recruitment of motor units.

Long-term potentiation of the human blink reflex.

The trigeminal reflex blink is an ideal system to investigate whether stimulus paradigms that produce long-term potentiation (LTP) in vitro modify motor learning in humans. Presentation of 12 trains of low-intensity, high-frequency stimuli (HFS) to the supraorbital branch of the trigeminal nerve (SO) modified subsequent reflex blinks of human subjects. When HFS occurred concurrently with reflex blinks, the procedure potentiated subsequent blinks for >1 hr. Combining HFS with feedback from the lid movement was critical for this facilitation because presenting HFS immediately after the blink did not alter subsequent blinks. When HFS preceded the blink, however, this treatment suppressed subsequent blinks for 30 min. These effects appear to occur within the trigeminal reflex blink circuits rather than at motoneurons, because stimulation of the previously HFS-treated SO evoked altered blinks in both eyelids, whereas stimulation of the untreated SO elicited unaltered blinks in both eyelids. The modified blink amplitude resulted from altering the response to A-fiber inputs to the trigeminal nerve because all stimuli were too weak to activate C-fibers. The data suggest that HFS produce LTP- and long-term depression (LTD)-like effects on wide dynamic range neurons in the trigeminal reflex blink circuit. The data also support the hypothesis that LTP and LTD mechanisms play a role in adaptive modification of human reflex blinks.

Aging of the trigeminal blink system.

This study characterizes trigeminal blinks in normal human subjects between 20 and 80 years of age, 60-year-old Parkinson's disease patients, and young and old guinea pigs. In normal humans over 60 years of age, lid-closing duration, and the excitability and latency of the trigeminal reflex blink increase significantly relative to younger subjects. Aged guinea pigs appear to display similar increases in reflex blink duration and latency. Reflex blink amplitude, however, does not change consistently with age. For subjects less than 70 years of age, a unilateral trigeminal stimulus evokes a 37% larger blink in the eyelid ipsilateral to the stimulus than in the contralateral eyelid, but 70-year-olds exhibit blinks of equal amplitude. In all cases, blink duration is identical for the two eyelids. If normal, age-related loss of dopamine neurons explains these trigeminal blink modifications, then Parkinson's disease should exaggerate age-related changes in these blink parameters. Preliminary data show that Parkinson's disease increases blink duration and excitability relative to age-matched control subjects. Thus, it seems likely that normal, age-related loss of dopamine neurons accounts for increases in trigeminal blink excitability and duration. A previously uncharacterized type of trigeminally evoked blink appears after age 40 in humans and in aged guinea pigs. In subjects less than 40 years old, a single trigeminal stimulus elicits a single reflex blink. In subjects over age 40, however, a single stimulus frequently evokes a reflex blink and additional blinks that occur at a fixed interval relative to the preceding blink. These "blink oscillations" may arise from oscillatory processes within trigeminal reflex blink circuits. The presence of exaggerated blink oscillations in subjects with dry eye and benign essential blepharospasm suggests that an alteration of blink oscillation mechanisms plays a critical role in these disorders.

Reflex excitability regulates prepulse inhibition.

Presentation of a weak stimulus, a prepulse, before a reflex-evoking stimulus decreases the amplitude of the reflex response relative to reflex amplitude evoked without a preceding prepulse. For example, presenting a brief tone before a trigeminal blink-eliciting stimulus significantly reduces reflex blink amplitude. A common explanation of such data are that sensory processing of the prepulse modifies reflex circuit behavior. The current study investigates the converse hypothesis that the intrinsic characteristics of the reflex circuit rather than prepulse processing determine prepulse modification of trigeminal and acoustic reflex blinks. Unilateral lesions of substantia nigra pars compacta neurons created rats with hyperexcitable trigeminal reflex blinks but normally excitable acoustic reflex blinks. In control rats, presentation of a prepulse reduced the amplitude of both trigeminal and acoustic reflex blinks. In 6-OHDA-lesioned rats, however, the same acoustic prepulse facilitated trigeminal reflex blinks but inhibited acoustic reflex blinks. The magnitude of prepulse modification correlated with reflex excitability. Humans exhibited the same pattern of prepulse modification. An acoustic prepulse facilitated the trigeminal reflex blinks of subjects with hyperexcitable trigeminal reflex blinks caused by Parkinson's disease. The same prepulse inhibited trigeminal reflex blinks of age-matched control subjects. Prepulse modification also correlated with trigeminal reflex blink excitability. These data show that reflex modification by a prepulse reflects the intrinsic characteristics of the reflex circuit rather than an external adjustment of the reflex circuit by the prepulse.

Identification and characterization of rat orbicularis oculi motoneurons using confocal laser scanning microscopy.

The eyeblink reflex is one of the most extensively studied behaviors in mammals. The active downward force that causes lid closure is controlled by the orbicularis oculi (OO) muscle. To augment our studies on the neurophysiology and plasticity of the rat eyeblink circuit, here we present the first anatomical paper to focus exclusively on identifying and characterizing the OO motoneurons of the rat facial motor nucleus (FMN). One thousand and twenty-nine cells from four animals were retrogradely labeled by injecting the OO muscle with HRP and were imaged conventionally. One hundred and one cells from five animals were labeled by injecting the OO muscle with a 3000 mol. wt. fluorescent dextran and were imaged using confocal laser scanning microscopy (CLSM). The latter method resulted in little tissue shrinkage, bright labeling, and excellent resolution of the soma, dendrites, and axon. Furthermore, it is a histologically simple alternative to HRP for retrograde labeling from the neuromuscular junction. Both methods revealed that the OO motoneurons were distributed over the entire length of the FMN, that they were concentrated along the dorsal crest of the nucleus, and that they were less numerous in the extreme rostral and caudal regions. As measured using the CLSM method, cell body areas were highly variable, ranging from 317 to 1500 microm2, but there was no size gradient along the rostrocaudal extent of the FMN. The neurons exhibited seven primary dendrites on average, which gave rise to bifurcating and even trifurcating secondary dendrites. Using the HRP method, the estimated area of OO motoneurons ranged from 161 to 1381 microm2. The combined methods furnished a detailed characterization of the number, spatial distribution, and morphology of rat OO motoneurons. Moreover, these methods provide a useful way to analyze the circuitry that modulates the rat eyeblink.

Animal model explains the origins of the cranial dystonia benign essential blepharospasm.

The current study demonstrates that combining two mild alterations to the rat trigeminal reflex blink system reproduces the symptoms of benign essential blepharospasm, a cranial dystonia characterized by uncontrollable spasms of blinking. The first modification, a small striatal dopamine depletion, reduces the tonic inhibition of trigeminal reflex blink circuits. The second alteration, a slight weakening of the lid-closing orbicularis oculi muscle, begins an adaptive increase in the drive on trigeminal sensory-motor blink circuits that initiates blepharospasm. By themselves, neither of these modifications causes spasms of lid closure, but combined, they induce bilateral forceful blinking and spasms of lid closure. A two-factor model based on these rodent experiments may explain the development of benign essential blepharospasm in humans. The first factor, a subclinical loss of striatal dopamine, creates a permissive environment within the trigeminal blink circuits. The second factor, an external ophthalmic insult, precipitates benign essential blepharospasm. This two-factor model may also be applicable to the genesis of other cranial dystonias.

Role of cerebellum in adaptive modification of reflex blinks.

We investigated the involvement of the cerebellar cortex in the adaptive modification of corneal reflex blinks and the regulation of normal trigeminal reflex blinks in rats. The ansiform Crus I region contained blink-related Purkinje cells that exhibited a complex spike 20.4 msec after a corneal stimulus and a burst of simple spike activity correlated with the termination of orbicularis oculi activity. This occurrence of the complex spike correlated with trigeminal sensory information associated with the blink-evoking stimulus, and the burst of simple spike activity correlated with sensory feedback about the occurrence of a blink. Inactivation of the inferior olive with lidocaine prevented all complex and significantly reduced simple spike modulation of blink-related Purkinje cells, but did not alter orbicularis oculi activity evoked by corneal stimulation. In contrast, both acute and chronic lesions of the cerebellar cortex containing blink-related Purkinje cells blocked adaptive increases in orbicularis oculi activity of the lid ipsilateral but not contralateral to the lesion. These data are consistent with the hypothesis that the cerebellum is part of a trigeminal reflex blink circuit. Changes in trigeminal signals produce modifications of the cerebellar cortex, which in turn, reinforce or stabilize modifications of brainstem blink circuits. When the trigeminal system does not attempt to alter the magnitude of trigeminal reflex blinks, cerebellar input has little or no effect on reflex blinks.

To blink or not to blink: inhibition and facilitation of reflex blinks.

A reflex blink typically inhibits subsequent blinks. In this study, we investigated whether the nature and time course of this inhibition vary when different combinations of blink-evoking stimuli are used. We used the paired stimulus paradigm, in which two blink-evoking stimuli-a conditioning stimulus followed by a test stimulus-are presented with a variety of interstimulus intervals, to examine the interactions between blinks evoked by trigeminal and acoustic stimuli in rats and humans. In addition, we studied the effect of a blink-evoking trigeminal stimulus on subsequent gaze-evoked blinks in humans. The results revealed that long-lasting inhibition occurred when the conditioning and test stimuli were within the same modality. A shorter period of inhibition followed by facilitation occurred when the stimuli were in different modalities. The data demonstrate that a blink-evoking stimulus initiates a lengthy period of inhibition in its own sensory pathway and a shorter period of inhibition in the reticular formation and/or in blink motoneurons. In addition, the results show that the blink-evoking stimulus also initiates a facilitatory process. Thus, the magnitude of a blink reflects a balance between inhibitory and facilitatory processes.

Influence of the superior colliculus on the primate blink reflex.

In this study we used microstimulation to investigate the influence of the superior colliculus on the trigeminal blink reflex. We report that stimulation in the intermediate to deep layers of the tectum produced inhibition of reflex blinks at a latency of approximately 26 ms. We considered the hypothesis that the blink inhibition was mediated via the omnipause neurons (OPNs) of the eye movement control system in the brainstem. Our results show that the least effective sites for suppression were in the rostral colliculus. This is inconsistent with the prediction that OPNs should be maximally recruited from the rostral tectum near the "fixation zone." From these points and other considerations, we conclude that the reflex blink suppression from the superior colliculus is not directly mediated by the OPNs or the saccadic eye movement circuits.

An explanation for reflex blink hyperexcitability in Parkinson's disease. I. Superior colliculus.

Hyperexcitable reflex blinks are a cardinal sign of Parkinson's disease. We investigated the neural circuit through which a loss of dopamine in the substantia nigra pars compacta (SNc) leads to increased reflex blink excitability. Through its inhibitory inputs to the thalamus, the basal ganglia could modulate the brainstem reflex blink circuits via descending cortical projections. Alternatively, with its inhibitory input to the superior colliculus, the basal ganglia could regulate brainstem reflex blink circuits via tecto-reticular projections. Our study demonstrated that the basal ganglia utilizes its GABAergic input to the superior colliculus to modulate reflex blinks. In rats with previous unilateral 6-hydroxydopamine (6-OHDA) lesions of the dopamine neurons of the SNc, we found that microinjections of bicuculline, a GABA antagonist, into the superior colliculus of both alert and anesthetized rats eliminated the reflex blink hyperexcitability associated with dopamine depletion. In normal, alert rats, decreasing the basal ganglia output to the superior colliculus by injecting muscimol, a GABA agonist, into the substantia nigra pars reticulata (SNr) markedly reduced blink amplitude. Finally, brief trains of microstimulation to the superior colliculus reduced blink amplitude. Histological analysis revealed that effective muscimol microinjection and microstimulation sites in the superior colliculus overlapped the nigrotectal projection from the basal ganglia. These data support models of Parkinsonian symtomatology that rely on changes in the inhibitory drive from basal ganglia output structures. Moreover, they support a model of Parkinsonian reflex blink hyper-excitability in which the SNr-SC target projection is critical.

An explanation for reflex blink hyperexcitability in Parkinson's disease. II. Nucleus raphe magnus.

Hyperexcitable reflex blinks are a cardinal sign of Parkinson's disease. The first step in the circuit linking the basal ganglia and brainstem reflex blink circuits is the inhibitory nigrostriatal pathway (Basso et al., 1996). The current study reports the circuits linking the superior colliculus (SC) to trigeminal reflex blink circuits. Microstimulation of the deep layers of the SC suppresses subsequent reflex blinks at a latency of 5.4 msec. This microstimulation does not activate periaqueductal gray antinociceptive circuits. The brainstem structure linking SC to reflex blink circuits must suppress reflex blinks at a shorter latency than the SC and produce the same effect on reflex blink circuits as SC stimulation, and removal of the structure must block SC modulation of reflex blinks. Only the nucleus raphe magnus (NRM) meets these requirements. NRM microstimulation suppresses reflex blinks with a latency of 4.4 msec. Like SC stimulation, NRM microstimulation reduces the responsiveness of the spinal trigeminal nucleus. Finally, blocking the receptors for the NRM transmitter serotonin eliminates SC modulation of reflex blinks, and muscimol inactivation of the NRM transiently prevents SC modulation of reflex blinks. Thus, the circuit through which the basal ganglia modulates reflex blinking is (1) the substantia nigra pars reticulata inhibits SC neurons, (2) the SC excites tonically active NRM neurons, and (3) NRM neurons inhibit spinal trigeminal neurons involved in reflex blink circuits.

The trigeminally evoked blink reflex. I. Neuronal circuits.

In this study, we characterized the pathways that generate the trigeminal blink reflex in the guinea pig. Blinks were evoked by stimulation of the supraorbital branch of the trigeminal nerve and measured by recording electromyographic activity in the lid-closing orbicularis oculi muscle (OOemg) and, in one case, lid position. Blinks evoked by stimulation of the supraorbital nerve consisted of two bursts of muscle activity ipsilateral to the side of stimulation. The first, R1, had a latency of 6.9 ms and the second, R2, had a latency of 17.25 ms. Increasing stimulus intensity to 3 times threshold for evoking an ipsilateral blink elicited an R1 and R2 response contralaterally, with latencies of 9.2 ms and 19.25 ms, respectively. We investigated the causes for this bipartite response that is seen in the guinea pig, as well as other mammals including humans. The two-component response could arise from different populations of afferents, or from different central circuits, or a combination of these two causes. Multiunit recording in the trigeminal ganglion and simultaneous measurement of the OOemg showed that activation of A beta afferents alone was sufficient to elicit both the R1 and the R2 responses, but that activation of A delta afferents could enhance both responses. Different neural circuits, however, produce the R1 and R2 responses. Transganglionic tracing with wheatgerm agglutin or choleragenoid subunit of cholera toxin bound to HRP revealed that primary afferents from the supraorbital branch of the trigeminal nerve terminated densely in the dorsal horn of spinal cord segment C1 and in the caudalis-interpolaris border region of the spinal trigeminal nucleus. Injections of HRP into the orbicularis oculi motoneuron region of the facial nucleus showed that both of these regions projected to the facial nucleus. Hemisections at the level of C1 eliminated the R2 blink response, but not the R1 response, evoked by stimulation of the supraorbital branch of the trigeminal nerve. Subsequent hemisections at the level of the obex eliminated the R1 response. Microinjections of the GABAB agonist baclofen into the spinal trigeminal nucleus at the level of the obex abolished the R1 but not the R2 response. Thus, the spinal trigeminal nucleus produces the R1 component, whereas the R2 component originates in the C1 region of the spinal cord.

The trigeminally evoked blink reflex. II. Mechanisms of paired-stimulus suppression.

The paired-stimulus paradigm, presentation of a pair of identical reflex-eliciting stimuli to the supraorbital nerve (SO) with an interstimulus interval of less than 2 s, evokes a response to the second, test, stimulus which is less than that elicited by the first, conditioning, stimulus. In this study, we investigated the site of this suppression and its pharmacology in the alert guinea pig. Both the early (R1) and the late (R2) component of the SO-evoked blink reflex exhibited suppression in the paired-stimulus paradigm. Initiation of suppression appeared to be specific to the afferent limb of the reflex rather than the result of motor activity generated by the conditioning stimulus. Neither acoustic conditioning stimuli nor air puffs that elicited blinks via another branch of the trigeminal nerve suppressed the test response. Extremely weak SO shocks, however, that did not directly elicit a reflex, caused suppression of the response to subsequent SO stimuli of normal intensity. Paired stimulus suppression of the R1 component appeared to involve activation of GABAB receptors within the spinal trigeminal nucleus. Both systemic injections and microinjections of baclofen into the spinal trigeminal nucleus enhanced R1 suppression, whereas identical injections of CGP35348, a GABAB antagonist, diminished R1 suppression. Furthermore, single-unit recordings in alert animals revealed that spinal trigeminal neurons exhibited suppression in the paired-stimulus paradigm that resembled that of the R1 component of the blink reflex. These findings showed that sensory gating underlies paired-stimulus suppression of the SO-evoked blink reflex and that activation of GABAB receptors plays an important role in this process.

The orientation of the cervical vertebral column in unrestrained awake animals. II. Movement strategies.

Previously we demonstrated a stereotyped resting posture of the head-neck arrangement in a number of vertebrates: the cervical vertebral column is oriented vertically to form one portion of the partial S-shaped configuration of the entire spine. The present investigation quantified the various strategies of head-neck movements employed by different mammalian species (human, monkeys, cats, rabbits and guinea pigs) using cineradiography. At rest, bipeds and quadrupeds hold their heads at the extreme point of flexion of the passive atlanto-occipital range of motion. In this posture, the horizontal semicircular canals are tilted upward from earth horizontal by 5 to 10 degrees and roughly parallel the plane determined by the two obliquus capitis posterior muscles. Furthermore, at this head position, the utricular maculae become oriented earth-horizontally. In quadrupedal animals, head-neck movements in the sagittal plane result from movement at the atlanto-occipital articulation (head/C1) and at the multi-articular cervico-thoracic junction (C6-Th3). Only very small flexion/extension movements occur within the body of the cervical vertebral column (C2-C5). Lowering the head from the resting position is only possible by flexion at the C6-Th3 vertebrae. Raising of gaze from the resting position is only possible by extension of the head at the atlanto-occipital articulation. By contrast, sagittal plane head movements in bipeds are largely confined to the cervico-thoracic junction. This is related to a significantly reduced range of motion of the atlanto-occipital articulation. In monkeys and humans, it range of motion is about 13 and 8-11 degrees, respectively, compared to 105 degrees in rabbits. Our cineradiographic data demonstrated different strategies for head movements in the sagittal plane between quadrupeds and bipeds. At one end of the spectrum, in the case of rabbits, there was no systematic relationship between head and neck orientation. Rabbits stabilized head posture by using the head-neck structure in a parallelogram fashion, which resulted in head posture being largely independent of cervical vertebral column orientation. In monkeys and humans, however, orientation of the head depended almost entirely on the orientation of the cervical vertebral column. In such case, head movements in the sagittal plane almost exclusively relied on the positioning of the cervico-thoracic junction. These different strategies again correlate with the different ranges of motion of the atlanto-occipital articulation. We suggest that vertebrates use mechanical constraints and preferred planes of action for head-neck movement control to simplify sensory-motor transformations subserving motor control and plasticity and to minimize neuronal operations.

Not looking while leaping: the linkage of blinking and saccadic gaze shifts.

Many vertebrates generate blinks as a component of saccadic gaze shifts. We investigated the nature of this linkage between saccades and blinking in normal humans. Activation of the orbicularis oculi, the lid closing muscle, EMG occurred with 97% of saccadic gaze shifts larger than 33 degrees. The blinks typically began simultaneously with the initiation of head and/or eye movement. To minimize the possibility that the blinks accompanying saccadic gaze shifts were reflex blinks evoked by the wind rushing across the cornea and eye-lashes as the head and eyes turned, the subjects made saccadic head turns with their eyes closed. In this condition, orbicularis oculi EMG activity occurred with all head turns greater than 17 degrees in amplitude and the EMG activity began an average of 39.3 ms before the start of the head movement. Thus, one component of the command for large saccadic gaze shifts appears to be a blink. We call these blinks gaze-evoked blinks. The linkage between saccadic gaze shifts and blinking is reciprocal. Evoking a reflex blink prior to initiating a voluntary saccadic gaze shift dramatically reduces the latency of the initiation of the head movement.

Pattern of extraocular muscle activation during reflex blinking.

Studies in humans and rabbits suggest that cocontraction of extraocular muscles occurs with reflex and voluntary blinks. We determined the pattern of extraocular muscle activity elicited by blink-evoking visual and trigeminal stimuli by electromyographically recording antagonistic pairs of extraocular muscles in alert rabbits. In addition, we recorded the activity of antidromically identified oculomotor motoneurons in response to the same stimuli in alert rabbits. The data demonstrate that all extraocular muscles except the superior oblique transiently increase their activity in response to blink-evoking stimuli. The pattern of extraocular muscle activity with reflex blinks mirrors that occurring in the lid-closing muscle, orbicularis oculi, but the latency of extraocular muscle activation is longer.

A role for the basal ganglia in nicotinic modulation of the blink reflex.

In humans and rats we found that nicotine transiently modifies the blink reflex. For blinks elicited by stimulation of the supraorbital branch of the trigeminal nerve, nicotine decreased the magnitude of the orbicularis oculi electromyogram (OOemg) and increased the latency of only the long-latency (R2) component. For blinks elicited by electrical stimulation of the cornea, nicotine decreased the magnitude and increased the latency of the single component of OOemg response. Since nicotine modified only one component of the supraorbitally elicited blink reflex, nicotine must act primarily on the central nervous system rather than at the muscle. The effects of nicotine could be caused by direct action on lower brainstem interneurons or indirectly by modulating descending systems impinging on blink interneurons. Since precollicular decerebration eliminated nicotine's effects on the blink reflex, nicotine must act through descending systems. Three lines of evidence suggest that nicotine affects the blink reflex through the basal ganglia by causing dopamine release in the striatum. First, stimulation of the substantia nigra mimicked the effects of nicotine on the blink reflex. Second, haloperidol, a dopamine (D2) receptor antagonist, blocked the effect of nicotine on the blink reflex. Third, apomorphine, a D2 receptor agonist, mimicked the effects of nicotine on the blink reflex.

Midbrain 6-hydroxydopamine lesions modulate blink reflex excitability.

The blink reflex abnormalities present in the 6 hydroxydopamine (6-OHDA) lesioned rat model of parkinsonism mimicked those of the human with Parkinson's disease. In alert rats, we monitored the long and short latency components of the orbicularis oculi electromyographic (OOemg) response evoked by electrical stimulation of the supraorbital branch of the trigeminal nerve (SO). Two paradigms, habituation and double pulse, provided a measure of blink reflex excitability. In normal rats, repeated stimulation of the SO produced habituation of the R2 component of the blink. In the double pulse paradigm, presentation of two identical SO stimuli resulted in a reduced or suppressed OOemg response to the second stimulus relative to the first. In rats with complete, unilateral lesions of midbrain dopamine neurons, repeated SO stimulation produced facilitation rather than habituation of the R2 component of the blink reflex. This facilitation occurred only with the eyelid contralateral to the lesion. In the double pulse paradigm, the lesioned rats showed increased excitability rather than suppression. This effect occurred bilaterally, although the increased excitability was strongest contralateral to the lesion. Rats with partial lesions of midbrain dopamine neurons exhibited qualitatively similar, but less pronounced blink reflex abnormalities. The R1 component of the blink reflex was unaffected by either the complete or partial lesions. Thus, modification of the blink reflex by 6-OHDA lesions provides a reproducible parkinsonian-like symptom which is amenable to investigations of increases in reflex excitability.

Botulinum toxin paralysis of the orbicularis oculi muscle. Types and time course of alterations in muscle structure, physiology and lid kinematics.

In chronically prepared guinea pigs, we investigated the time course of botulinum toxin A's (Bot A) effect on the blink reflex by monitoring lid movements and EMG activity prior to and after Bot A injection into the orbicularis oculi muscle (OOemg), or after nerve crush of the zygomatic nerve. We correlated these alterations with the morphological changes of the orbicularis oculi (lid-closing) muscles of the same animals. After Bot A treatment there was a profound reduction of OOemg activity and blink amplitudes as well as a slowing of maximum blink down-phase velocity. Blink up-phases, however, remained unchanged. Gradual recovery of OOemg magnitude and blink amplitude started around day 6; a functioning blink reflex appeared on day 21, and full recovery of blink amplitude occurred by day 42. Crushing the zygomatic branch of the facial nerve produced similar changes in blink parameters, but recovery was much more rapid (15 days) than for Bot A-treated guinea pigs. The morphological analysis demonstrated that Bot A produced a denervation-like atrophy in the orbicularis oculi. No fiber type-specific alterations were noted, and all muscle fiber types ultimately recovered, with no longstanding consequences of the transient denervation. Our findings support the notion that functional recovery was the result of preterminal and terminal axonal sprouting that subsequently re-established functional innervation. Moreover, differences between the present findings and those seen after injection of Bot A into the extraocular muscles strongly support the hypothesis that the composition in terms of muscle fiber type and the properties of the motor control system of a given muscle greatly influence both how the particular muscle responds to toxin injection, and how effective the toxin is in resolution of neuromuscular disorders that affect a particular muscle. The present findings were consistent with clinical observations that Bot A produces only temporary relief in patients with essential blepharospasm. It is likely that the efficacy of Bot A in treatment of blepharospasm could be improved by using agents that suppress terminal sprouting. The close correspondence of the changes in blink physiology between human patients and guinea pigs after Bot A treatment demonstrate that the guinea pig is an excellent model system for testing strategies to prolong the beneficial effects of Bot A treatment in relieving lid spasms in human subjects.