Paola D'Ascanio

Motoneurons of twitch and nontwitch extraocular muscle fibers in the abducens, trochlear, and oculomotor nuclei of monkeys

Eye muscle fibers can be divided into two categories: nontwitch, multiply innervated muscle fibers (MIFs), and twitch, singly innervated muscle fibers (SIFs). We investigated the location of motoneurons supplying SIFs and MIFs in the six extraocular muscles of monkeys. Injections of retrograde tracers into eye muscles were placed either centrally, within the central SIF endplate zone; in an intermediate zone, outside the SIF endplate zone, targeting MIF endplates along the length of muscle fiber; or distally, into the myotendinous junction containing palisade endings. Central injections labeled large motoneurons within the abducens, trochlear or oculomotor nucleus, and smaller motoneurons lying mainly around the periphery of the motor nuclei. Intermediate injections labeled some large motoneurons within the motor nuclei but also labeled many peripheral motoneurons. Distal injections labeled small and medium‐large peripheral neurons strongly and almost exclusively. The peripheral neurons labeled from the lateral rectus muscle surround the medial half of the abducens nucleus: from superior oblique, they form a cap over the dorsal trochlear nucleus; from inferior oblique and superior rectus, they are scattered bilaterally around the midline, between the oculomotor nucleus; from both medial and inferior rectus, they lie mainly in the C‐group, on the dorsomedial border of oculomotor nucleus. In the medial rectus distal injections, a “C‐group extension” extended up to the Edinger‐Westphal nucleus and labeled dendrites within the supraoculomotor area. We conclude that large motoneurons within the motor nuclei innervate twitch fibers, whereas smaller motoneurons around the periphery innervate nontwitch, MIF fibers. The peripheral subgroups also contain medium‐large neurons which may be associated with the palisade endings of global MIFs. The role of MIFs in eye movements is unclear, but the concept of a final common pathway must now be reconsidered. J. Comp. Neurol. 438:318–335, 2001.

C-Fos expression in the rat brain after unilateral labyrinthectomy and its relation to the uncompensated and compensated stages

The expression of the immediate early gene c-fos has been studied in the entire brain of rats 3, 6 and 24 h after surgical unilateral labyrinthectomy. We combined in situ hybridization for c-fos messenger RNA with immunocytochemistry for Fos protein to document very early changes in c-fos expression and to identify with cellular resolution neuronal populations activated by unilateral labyrinthectomy. Three hours after unilateral labyrinthectomy a bilateral increase in both c-fos messenger RNA and protein levels was seen in the superior, medial and spinal vestibular nuclei, nucleus Y, and prepositus hypoglossal nucleus. These changes were asymmetric in the medial vestibular nucleus, being most prominent in the dorsal part of the contralateral nucleus (where second order vestibular neurons are located) and in the ventral part of the ipsilateral nucleus (where commissural neurons acting on the medial vestibular nucleus of the intact side are located). An increase in c-fos messenger RNA expression was seen bilaterally, but with an ipsilateral predominance, in the vermal and paravermal areas of the cerebellar cortex, flocculus and paraflocculus, as well as in the precerebellar lateral and paramedian reticular nuclei. c-fos messenger RNA and protein levels increased in a few regions of the contralateral inferior olive. A predominantly ipsilateral increase in c-fos expression also occurred in the caudate-putamen. A bilateral but not exactly symmetric increase in both c-fos messenger RNA and protein levels was present in several nuclei of the dorsal pontine tegmentum (parabrachial nucleus, locus coeruleus and laterodorsal tegmental nucleus), mesencephalic periaqueductal gray, and several hypothalamic, thalamic and cerebrocortical regions. No change was seen in the cerebellar nuclei, lateral vestibular nucleus and red nucleus. The increased expression of c-fos observed 3 h after unilateral labyrinthectomy, in conjunction with the sudden occurrence of postural and motor deficits, usually declined 6-24 h after the lesion, i.e. during the development of vestibular compensation. In the dorsal part of the medial vestibular nucleus, however, the pattern of c-fos expression observed 3 h after unilateral labyrinthectomy was reversed 6-24 h after the lesion: both c-fos messenger RNA and protein levels increased on the ipsilateral side, but greatly decreased on the contralateral side. In conclusion, asymmetric changes in c-fos expression occurred within 3 h after unilateral labyrinthectomy, but gradually declined or reversed 6 and 24 h after the lesion, thus being temporally related to the appearance and development of vestibular compensation.

Responses of locus coeruleus and subcoeruleus neurons to sinusoidal stimulation of labyrinth receptors

In precollicular decerebrate cats the electrical activity of 141 individual neurons located in the locus coeruleus-complex, i.e. in the dorsal (n = 41) and ventral parts (n = 67) as well as in the locus subcoeruleus (n = 33), was recorded during sinusoidal tilt about the longitudinal axis of the whole animal, leading to stimulation of labyrinth receptors. Some of these neurons showed physiological characteristics attributed to the norepinephrine-containing locus coeruleus neurons, namely, (i) a slow and regular resting discharge, and (ii) a typical biphasic response to fore- and hindpaw compression consisting of short impulse bursts followed by a silent period, which has been attributed to recurrent and/or lateral inhibition of the norepinephrine-containing neurons. Furthermore, 16 out of the 141 neurons were activated antidromically by stimulation of the spinal cord at T12 and L1, thus being considered coeruleospinal or subcoeruleospinal neurons. A large number of tested neurons (80 out of 141, i.e. 56.7%) responded to animal rotation at the standard frequency of 0.15 Hz and at the peak amplitude of 10 degrees. However, the proportion of responsive neurons was higher in the locus subcoeruleus (72.7%) and the dorsal locus coeruleus (61.0%) than in the ventral locus coeruleus (46.3%). A periodic modulation of firing rate of the units was observed during the sinusoidal stimulus. In particular, 45 out of the 80 units (i.e. 56.2%) were excited during side-up and depressed during side-down tilt (beta-responses), whereas 20 of 80 units (i.e. 25.0%) showed the opposite behavior (alpha-responses). In both instances, the response peak occurred with an average phase lead of about + 18 degrees, with respect to the extreme side-up or side-down position of the animal; however, the response gain (imp./s per deg) was, on average, more than two-fold higher in the former than in the latter group. The remaining 15 units (i.e. 18.7%) showed a prominent phase shift of this response peak with respect to animal position. Similar results were obtained from the subpopulation of locus coeruleus-complex neurons which fired at a low rate (less than 5.0 imp./s), as well as for the antidromically identified coeruleospinal neurons. The response gain of locus coeruleus-complex neurons, including the coeruleospinal neurons, did not change when the peak amplitude of tilt was increased from 5 degrees to 20 degrees at the fixed frequency of 0.15 Hz. This indicates that the system was relatively linear with respect to the amplitude of displacement.(ABSTRACT TRUNCATED AT 400 WORDS)

Functional Structure of Spontaneous Sleep Slow Oscillation Activity in Humans

Background During non-rapid eye movement (NREM) sleep synchronous neural oscillations between neural silence (down state) and neural activity (up state) occur. Sleep Slow Oscillations (SSOs) events are their EEG correlates. Each event has an origin site and propagates sweeping the scalp. While recent findings suggest a SSO key role in memory consolidation processes, the structure and the propagation of individual SSO events, as well as their modulation by sleep stages and cortical areas have not been well characterized so far. Methodology/Principal Findings We detected SSO events in EEG recordings and we defined and measured a set of features corresponding to both wave shapes and event propagations. We found that a typical SSO shape has a transition to down state, which is steeper than the following transition from down to up state. We show that during SWS SSOs are larger and more locally synchronized, but less likely to propagate across the cortex, compared to NREM stage 2. Also, the detection number of SSOs as well as their amplitudes and slopes, are greatest in the frontal regions. Although derived from a small sample, this characterization provides a preliminary reference about SSO activity in healthy subjects for 32-channel sleep recordings. Conclusions/Significance This work gives a quantitative picture of spontaneous SSO activity during NREM sleep: we unveil how SSO features are modulated by sleep stage, site of origin and detection location of the waves. Our measures on SSOs shape indicate that, as in animal models, onsets of silent states are more synchronized than those of neural firing. The differences between sleep stages could be related to the reduction of arousal system activity and to the breakdown of functional connectivity. The frontal SSO prevalence could be related to a greater homeostatic need of the heteromodal association cortices.

Spinal Projections from the Mesencephalon in the Toad

Mesencephalic cell groups projecting to the spinal cord have been identified by means of the retrograde axonal transport of the enzyme horseradish peroxidase (HRP). The injections were made either in the cervical or lumbar enlargements of the toad spinal cord. Following injections in the cervical cord, labeled cells are located in the isthmus region, in the ipsilateral laminated nucleus posteroventralis tegmenti mesencephali (Potter). At more rostral levels the labeled cells are in the nucleus of the fasciculus longitudinalis medialis, in the nucleus interstitialis of the fasciculus longitudinalis medialis, in the contralateral red nucleus, in lamina six of the contralateral optic tectum, bilaterally in the nucleus of the posterior commissure and in the mesencephalic nucleus in the Vth nerve. Injections in the lumbar cord label neurons of the nucleus posteroventralis tegmenti mesencephali (Potter) and nucleus interstitialis of the fasciculus longitudinalis medialis. Nuclei that had not been previously identified in anurans but which were labeled after HRP spinal injections (i.e., the nucleus interstitialis of the fasciculus longitudinalis medialis, the nucleus of the posterior commissure and the red nucleus) have been delimited in normal material in Nissl-stained transverse sections. The spinal pathways from the mesencephalon can be classified into four projections: reticulospinal, rubrospinal, tectospinal and trigeminospinal. A comparison of these descending fiber systems with homologous pathways in other vertebrate species has been made.

Locus coeruleus and dorsal pontine reticular influences on the gain of vestibulospinal reflexes

Experimental anatomical and physiological studies have shown that noradrenergic locus coeruleus (LC) neurons, which are NE-sensitive due to inhibitory adrenoceptors, send inhibitory afferents to neurons of the peri-LC alpha and the adjacent dorsal pontine reticular formation (pRF); on the other hand these tegmental neurons, which are, in part at least, cholinergic as well as cholinoceptive, send excitatory afferents to the medullary inhibitory reticulospinal (RS) system. Experiments performed in precollicular decerebrate cats indicate that these pontine structures exert a regulatory influence on posture as well as on the gain of vestibulospinal (VS) reflexes. In particular, the increased discharge of dorsal pontine reticular neurons, and the related inhibitory RS neurons induced by microinjection of cholinergic agonists into the peri-LC alpha and the adjacent pRF of one side, decreased the postural activity, but greatly increased the response gain of the ipsilateral triceps brachii in response to stimulation of labyrinth receptors resulting from roll tilt of the animal (at 0.15 Hz, +/- 10 degrees). Similar results were also obtained when the discharge of these pontine and medullary reticular neurons was raised, either by local injection into the peri-LC alpha and the dorsal pRF of the beta-adrenergic antagonist propranolol, which blocked the inhibitory influence of the noradrenergic LC neurons on these structures, or by local injection into the LC complex of an alpha 2- or beta-adrenergic agonist (clonidine or isoproterenol) which led to functional inactivation of the noradrenergic neurons; in the latter case the effects were bilateral. Just the opposite results were obtained after microinjection into the LC of a cholinergic agonist, leading to activation of the corresponding neurons. Evidence was also presented indicating that the cholinergic excitatory afferents to the LC originated from the ipsilateral dorsal pRF. The effects described above were dose-dependent and site-specific, as shown by histological controls. Under given conditions, the decrease in postural activity induced either by direct activation of presumptive cholinergic and cholinoceptive pRF neurons or by inactivation of noradrenergic and NE-sensitive LC neurons was followed by transient episodes of postural atonia which lasted several minutes and affected the ipsilateral and sometimes also the contralateral limbs. In these instances, the EMG modulation of the corresponding triceps brachii to animal tilt was suppressed. These findings suggest two different ranges of operation for the noradrenergic and cholinergic structures located in the dorsolateral pontine tegmentum, leading either to a decrease or to an increase in gain of the VS reflexes. The cellular basis of these gain changes is discussed.(ABSTRACT TRUNCATED AT 400 WORDS)

Gene expression in autonomic areas of the medulla and the central nucleus of the amygdala in rats during and after space flight

During space flight astronauts show vestibular-related changes in balance, eye movements, and spontaneous and reflex control of cardiovascular, respiratory and gastrointestinal function, sometimes associated with space motion sickness. These symptoms undergo compensation over time. Here we used changes in the expression of two immediate-early gene (IEG) products to identify cellular and molecular changes occurring in autonomic brainstem regions of adult male albino rats killed at different times during the Neurolab Space Mission (STS-90). Both direct effects of gravitational changes, as well as indirect effects of gravitational changes on responses to light exposure were examined. Regions under the direct control of vestibular afferents such as the area postrema and the caudal part of the nucleus of the tractus solitarius (NTSC) were both directly and indirectly affected by gravity changes. These areas showed no changes in the expression of IEG products during exposure to microgravity with respect to ground controls, but did show a significant increase 24 h after return to 1 G (gravity). Exposure to microgravity significantly inhibited gene responses to light exposure seen after return to 1 G. A similar direct and indirect response pattern was also shown by the central nucleus of the amygdala, a basal forebrain structure anatomically and functionally related to the NTS. The rostral part of the NTS (NTSR) receives different afferent projections than the NTSC. This region did not show any direct gravity-related changes in IEG expression, but showed an indirect effect of gravity on IEG responses to light. A similar pattern was also obtained in the intermediate reticular nucleus and the parvocellular reticular nucleus. Two other medullary reticular structures, the dorsal and the ventral medullary reticular nuclei showed a less well defined pattern of responses that differed from those seen in the NTSC and NTSR. The short- and long-lasting molecular changes in medullary and basal forebrain gene expression described here are thought to play an important role in the integration of autonomic and vestibular signals that ultimately regulate neural adaptations to space flight.

Responses of locus coeruleus and subcoeruleus neurons to sinusoidal neck rotation in decerebrate cat

The electrical activity of 99 neurons located in the locus coeruleus-complex, namely in the dorsal (n = 26) and the ventral part of the locus coeruleus (n = 46) as well as the locus subcoeruleus (n = 27), has been recorded in precollicular decerebrate cats during sinusoidal displacement of the neck. This was achieved by rotation of the body about the longitudinal axis of the animal, while maintaining the head stationary. A proportion of these neurons showed some of the main physiological characteristics attributed to the noradrenergic locus coeruleus neurons, i.e. (i) a slow and regular resting discharge, and (ii) a typical biphasic response to fore and hindpaw compression consisting of short bursts of impulses followed by a period of quiescence, due at least in part to recurrent or lateral inhibition of the corresponding neurons. Moreover, 14 out of the 99 neurons were activated antidromically by stimulation of the spinal cord at T12 and L1, thus being considered as coeruleo- or subcoeruleospinal neurons. Among these locus coeruleus-complex neurons tested, 73 out of 99 (i.e. 73.7%) responded to neck rotation at the standard frequency of 0.15 Hz and at the peak amplitude of displacement of 10 degrees. In particular 40 of 73 units (i.e. 54.8%) were excited during side-down neck rotation and depressed during side-up rotation, while 18 of 73 units (i.e. 24.7%) showed the opposite pattern. In both instances the peak of the responses occurred with an average phase lead of +34.2 degrees for the extreme side-down or side-up neck displacement; however, the response gain (impulses/s per deg) was on the average more than two-fold higher in the former than in the latter group of units. The remaining 15 units (i.e. 20.5%) showed phase angle values which were intermediate between those of the two main populations. As to the coeruleo or subcoeruleospinal neurons, 11 of 14 units (78.6%) responded to the neck input, the majority (nine of 11 units, i.e. 81.8%) being excited during side-down neck rotation. Within the explored region, the proportion of responsive units was higher in the locus subcoeruleus (85.2%) than in the locus coeruleus, both dorsal and ventral (69.4%). Moreover, units located in the former structure showed on the average a response gain higher than that found in the latter structures. Similar results were also obtained from the population of locus subcoeruleus-complex neurons which fired at a low rate (less than or equal to 5.0 impulses/s).(ABSTRACT TRUNCATED AT 400 WORDS)

Convergence and interaction of neck and macular vestibular inputs on locus coeruleus and subcoeruleus neurons.

Extracellular recordings were obtained in precollicular decerebrate cats from 90 neurons located in the noradrenergic area of the dorsal pontine tegmentum, namely in the dorsal (LCd,n=24) and the ventral part (LCα,n=40) of the locus coeruleus (LC) as well as in the locus subcoeruleus (SC,n=26). Among these units of the LC complex, 13 were coerulospinal (CS) neurons antidromically identified following stimulation of the spinal cord at T12-L1. Some of these neurons showed the main physiological characteristics of the norepinephrine (NE)-containing LC neurons, i.e., a slow and regular resting discharge and a typical biphasic response to fore- and hindpaw compression consisting of a short burst of excitation followed by a period of quiescence, due, in part at least, to recurrent and/or lateral inhibition. Unit firing rate was analyzed under separate stimulation of macular vestibular, neck, or combined receptors by using sinusoidal rotation about the longitudinal axis at 0.15 Hz, ±10° peak amplitude.Among the 90 LC-complex neurons, 60 (66.7%) responded with a periodic modulation of their firing rate to roll tilt of the animal and 67 (74.4%) responded to neck rotation. Convergence of macular and neck inputs was found in 52/90 (57.8%) LC-complex neurons; in these units, the gain and the sensitivity of the first harmonic of the response corresponded on the average to 0.34±0.45, SD, impulsed·s−1·deg−1 and 3.55±2.82, SD, %/deg for the neck responses and to 0.23±0.29, SD, impulses·s−1·deg−1 and 3.13±3.04, SD, %/deg for the macular responses. In addition to these convergent units, 8/90 (8.9%) and 15/90 (16.7%) LC-complex units responded to selective stimulation either of macular or of neck receptors only. These units displayed a significantly lower response gain and sensitivity to animal tilt and neck rotation with respect to those obtained from convergent units. Most of the convergent LC-complex units were maximally excited by the direction of stimulus orientation, the first harmonic of responses showing an average phase lead of about +31.0° with respect to neck position and +17.6° with respect to animal position. Two populations of convergent neurons were observed. The first group of units (43/52, i.e., 82.7%) showed reciprocal (“out of phase” responses to the two inputs; moreover, most of these units were excited during side-down neck rotation, but inhibited during side-down animal tilt. The second group of units (9/52, i.e., 17.3%) showed parallel (“in phase” responses to the two inputs and they were excited by side-down or side-up neck rotation and animal tilt. The response characteristics of LC-complex neurons to combined neck and macular inputs, elicited during head rotation, corresponded to those predicted by a vectorial summation of the individual neck and macular responses. In particular, “out of phase” units displayed small amplitudes and large phase shifts of their responses with respect to those obtained during individual neck or macular stimulation. In contrast, “in phase” units displayed large responses during head rotation. Some nonlinearities of the responses to combined stimulation of neck and macular receptors, however, were observed. The possibility that the CS neurons contributed, with the vestibulospinal (VS) neurons, to the postural adjustments of the limb musculature during labyrinth and neck reflexes was discussed.

Likeness-Based Detection of Sleep Slow Oscillations in Normal and Altered Sleep Conditions: Application on Low-Density EEG Recordings

Sleep slow oscillation (SSO) is a common EEG pattern of spontaneous activity during nonrapid eye movement sleep. A new method for detecting SSOs is presented and compared to previous canonical methods. The main result of this research is that for the first time an extensive SSO analysis is applied to clinical EEG montages, based on low-density EEG recordings. The proposed method gives positive indications about its effectiveness also for altered sleep, extending the SSO analysis to extreme cases, and thus, opening a new front for investigating pathophysiological correlates of sleep disorders.