Neuronal activity in the medulla oblongata during vocalization. A single-unit recording study in the squirrel monkey

In six squirrel monkeys (Saimiri sciureus), the medulla oblongata was explored with microelectrodes, looking for vocalization-correlated activity. The vocalizations were elicited by microinjections of glutamate agonists into the periaqueductal grey of the midbrain. Vocalization-related cells were found in greater numbers in the nucl. ambiguus (Ab) and retroambiguus (RAb), in the parvocellular, magnocellular and central reticular formation as well as in the solitary tract nucleus and spinal trigeminal nucleus. Small numbers were also found in the vestibular complex, cuneate nuclei, inferior olive and lateral reticular nucleus. A differentiation of the neuronal responses into 12 reaction types reveals that the frequency of each reaction type varies from brain structure to brain structure, thus allowing a specification of the different vocalization-related areas. According to this specification, it is proposed that initiation of vocalization takes place via the parvocellular reticular formation; vocal pattern control is mainly brought about by the parvocellular reticular formation, Ab, solitary tract nucleus and spinal trigeminal nucleus; expiratory control and respiratory-laryngeal coordination is carried out by the RAb, Ab and central nucleus of the reticular formation; vocalization-specific postural adjustments are carried out via the vestibular and cuneate nuclei.     

Neuronal activity in the inferior colliculus and bordering structures during vocalization in the squirrel monkey

In four squirrel monkeys (Saimiri sciureus), the inferior colliculus, together with the neighboring superior colliculus, reticular formation, cuneiform nucleus and parabrachial area, were explored with microelectrodes, looking for neurons that might be involved in the discrimination between self-produced and external sounds. Vocalization was elicited by kainic acid injections into the periaqueductal gray of the midbrain. Acoustic tests were carried out with ascending and descending narrow-band noise sweeps spanning virtually the whole hearing range of the squirrel monkey. Altogether 577 neurons were analyzed. Neurons that both were audiosensitive and fired in advance of self-produced vocalization were found almost exclusively in the pericentral nuclei of the inferior colliculus and the adjacent reticular formation. Only the latter, however, contained, in addition, neurons that fired during external acoustic stimulation, but remained quiet during self-produced vocalization. These findings suggest that the reticular formation bordering the inferior colliculus is involved in the discrimination between self-produced and foreign vocalization on the basis of a vocalmotor feedforward mechanism.     

2-Deoxyglucose uptake during vocalization in the squirrel monkey brain

In the squirrel monkey (Saimiri sciureus), the cerebral 2-deoxyglucose uptake was compared between animals made to vocalize by electrical stimulation of the periaqueductal grey and animals stimulated in the same structure, but sub-threshold for vocalization. A significantly higher 2-deoxyglucose uptake in the vocalizers than the non-vocalizers was found in the dorsolateral prefrontal cortex, supplementary and pre-supplementary motor area, anterior and posterior cingulate cortex, primary motor cortex, claustrum, centrum medianum, perifornical hypothalamus, periaqueductal grey, intercollicular region, dorsal mesencephalic reticular formation, peripeduncular nucleus, substantia nigra, nucl. ruber, paralemniscal area, trigeminal motor, principal and spinal nuclei, solitary tract nucleus, nucl. ambiguus, nucl. retroambiguus, nucl. hypoglossus, ventral raphe and large parts of the medullary reticular formation. The study makes clear that vocalization, even in the case of genetically pre-programmed patterns, depends upon an extensive network, beyond the well-known periaqueductal grey, nucl. retroambiguus and cranial motor nuclei pathway.     

The effects of brainstem lesions on vocalization in the squirrel monkey

The present study is an attempt to find out the brain areas involved in the motor coordination of species-specific vocalization. For this purpose, high-frequency coagulations were placed in a systematic manner throughout the brainstem and posterior diencephalon in altogether 43 squirrel monkeys (Saimiri sciureus). The effect of these lesions on different call types elicited by electrical brain stimulation was studied spectrographically. It was found that bilateral destruction of the ventrolateral, ventroposterior and intralaminar thalamus, periventricular and rostral periaqueductal gray, ventral tegmental area of Tsai, nucl. interpeduncularis, nucl. ruber, anterodorsolateral midbrain tegmentum, superior and inferior colliculi, pontine gray, cerebral peduncles, medial pontine reticular formation, raphe and vestibular nuclei did not affect the acoustic structure of the calls tested. On the other hand, lesions in the ventrolateral midbrain involving the substantia nigra and overlying reticular formation, in the midbrain tegmentum just below the inferior colliculus, in the lateral pons and almost the whole medulla (minimal lesion size: 2.5 mm3) changed vocalization significantly. It is suggested that the latter areas are more or less directly involved in the motor coordination of vocalization, while the first are not.     

On the role of the reticular formation in vocal pattern generation

This review is an attempt to localize the brain region responsible for pattern generation of species-specific vocalizations. A catalogue is set up, listing the criteria considered to be essential for a vocal pattern generator. According to this catalogue, a vocal pattern generator should show vocalization-correlated activity, starting before vocal onset and reflecting specific acoustic features of the vocalization. Artificial activation by electrical or glutamatergic stimulation should produce artificially sounding vocalization. Lesioning is expected to have an inhibitory or deteriorating effect on vocalization. Anatomically, a vocal pattern generator can be assumed to have direct or, at least, oligosynaptic connections with all the motoneuron pools involved in phonation. A survey of the literature reveals that the only area meeting all these criteria is a region, reaching from the parvocellular pontine reticular formation just above the superior olive through the lateral reticular formation around the facial nucleus and nucleus ambiguus down to the caudalmost medulla, including the dorsal and ventral reticular nuclei and nucleus retroambiguus. It is proposed that vocal pattern generation takes place within this whole region.     

The effects of lesions of medullary midline and lateral reticular areas on inhibition in the dorsal horn produced by periaqueductal grey stimulation in the cat

In barbiturate-anaesthetized cats, the excitation of lumbar dorsal horn neurones by impulses in unmyelinated primary afferent fibres was inhibited by electrical stimulation in the periaqueductal grey matter. This inhibition was slightly reduced by extensive electro-coagulation of the medullary midline and para-medial areas including the raphe´, but significantly reduced by small bilateral lesions in the region of the caudal lateral reticular nuclei. When the lateral lesions were made subsequent to midline coagulation, the inhibition from periaqueductal grey stimulation was abolished. An important component of spinal inhibition from periaqueductal grey stimulation appears to relay in lateral reticular areas of the medulla.     

The central mesencephalic grey in birds: nucleus intercollicularis and substantia grisea centralis

The question discussed in this paper is, whether the dorsomedial part of the intercollicular nucleus and central mesencephalic grey of birds are comparable to (parts of) the periaqueductal grey in mammals. The mammalian periaqueductal grey, and the avian dorsomedial part of the intercollicular nucleus + central mesencephalic grey are each part of pathways in control of functions such as vocalization and sexual behavior. The connectivity and histochemical features of the dorsomedial intercollicular nucleus and central mesencephalic grey are partly different and also differ partly from those of the mammalian periaqueductal grey. It is suggested that these areas in mammals and birds form comparable links in the emotional motor pathway that has been defined before in mammals.     

Brain afferents to the medullary dorsal reticular nucleus: a retrograde and anterograde tracing study in the rat

The medullary dorsal reticular nucleus (DRt) was recently shown to belong to the supraspinal pain control system; neurons within this nucleus give origin to a descending projection that increases spinal nociceptive transmission and facilitates pain perception [Almeida et al. (1999), Eur. J. Neurosci., 11, 110-122]. In the present study, the areas of the brain that may modulate the activity of DRt neurons were investigated by using of tract-tracing techniques. Injection of a retrograde tracer into the DRt resulted in labelling in multiple areas of the brain. In the contralateral orbital, prelimbic, infralimbic, insular, motor and somatosensory cortices labelling was prominent, but a smaller ipsilateral projection from these same areas was also detected. Strong labelling was also noted in the central amygdaloid nucleus, bed nucleus of stria terminalis and substantia innominata. Labelled diencephalic areas were mainly confined to the hypothalamus, namely its lateral and posterior areas as well as the paraventricular nucleus. In the mesencephalon, the periaqueductal grey, red nucleus and deep mesencephalic nucleus were strongly labelled, whereas, in the brainstem, the parabrachial nuclei, rostroventromedial medulla, nucleus tractus solitarius, spinal trigeminal nucleus, and the parvocellular, dorsal, lateral and ventral reticular nuclei were the most densely labelled regions. All deep cerebellar nuclei were labelled bilaterally. These data suggest that the DRt integrates information from the somatosensory, antinociceptive, autonomic, limbic, pyramidal and extrapyramidal systems while triggering its descending facilitating action upon the spinal nociceptive transmission.     

Glutamate-induced vocalization in the squirrel monkey

In the squirrel monkey, 164 brain sites yielding vocalization when electrically stimulated were tested for their capability to produce vocalization when injected with mono-sodium-L-glutamate. The sites were located in the anterior limbic cortex, n. accumbens, substantia innominata, amygdala, n. striae terminalis, hypothalamus, midline thalamus, field H of Forel, substantia nigra, periventricular and periaqueductal gray, inferior colliculus, reticular formation of midbrain, pons and medulla, inferior olive, lateral reticular nucleus and nucleus of solitary tract. Of the 164 sites tested, 31 were positive. These were located in the substantia innominata, caudal periventricular and periaqueductal gray, lateral pontine and medullary reticular formation. While all the calls obtained from the forebrain and midbrain had a normal acoustic structure, most pontine and all medullary vocalizations had an artificial character. It is concluded that: the substantia innominata, caudal periventricular and periaqueductal gray, lateral pontine and medullary reticular formation represent relay stations of vocalization-controlling pathways; the periaqueductal gray represents the lowest relay station above the level of motor coordination; neurons responsible for motor coordination of vocalization lie in the reticular formation around the caudal brachium conjunctivum, the superior olive, n. facialis, n. ambiguus and below the n. solitarius; not all areas from which vocalization can be obtained by electrical stimulation of nerve cell bodies, dendrites and nerve endings (in contrast to fibers en passage) also yield vocalization when stimulated with glutamate.     

The projections of the dorsomedial hypothalamic nucleus in the rat

The dorsomedial hypothalamic nucleus (DMH) output pathways are revealed by using autoradiographic tracing of tritium labeled Leucine and by the recently introduced Phaseolus vulgaris leuco-agglutinin immunocytochemical method. Terminal labeling appears in the dorsal motor nucleus of the vagus, nucleus ambiguus and in the parvocellular reticular formation at the lower medullary level. Mesencephalic labeling is found in the periaqueductal gray at the level of the oculomotor nucleus. In the hypothalamus labeled terminal boutons are identified in the lateral and ventromedial hypothalamic nuclei but also in the parvocellular paraventricular nucleus. Furthermore, the circumventricular organs are found to receive a dense DMH input, particularly the organum vasculosum of the lamina terminalis and the subfornical organ. These findings are discussed in relation to the dorsomedial nucleus involvement in the control of feeding and pancreatic hormone release. It appears that the DMH participates in this control via descending pathways to the preganglionic pancreas innervating neurons but also via a neuroendocrine route. The latter connection is indicated by terminal labeling in the parvocellular paraventricular nucleus in the area that contains the corticotropin-releasing factor positive cells.     

Functional circuitry involved in the regulation of whisker movements.

Neuroanatomical tract-tracing methods were used to identify the oligosynaptic circuitry by which the whisker representation of the motor cortex (wMCx) influences the facial motoneurons that control whisking activity (wFMNs). Injections of the retrograde tracer cholera toxin subunit B into physiologically identified wFMNs in the lateral facial nucleus resulted in dense, bilateral labeling throughout the brainstem reticular formation and in the ambiguus nucleus as well as predominantly ipsilateral labeling in the paralemniscal, pedunculopontine tegmental, K?lliker-Fuse, and parabrachial nuclei. In addition, neurons in the following midbrain regions projected to the wFMNs: superior colliculus, red nucleus, periaqueductal gray, mesencephalon, pons, and several nuclei involved in oculomotor behaviors. Injections of the anterograde tracer biotinylated dextran amine into the wMCx revealed direct projections to the brainstem reticular formation as well as multiple brainstem and midbrain structures shown to project to the wFMNs. Regions in which retrograde labeling and anterograde labeling overlap most extensively include the brainstem parvocellular, gigantocellular, intermediate, and medullary (dorsal and ventral) reticular formations; ambiguus nucleus; and midbrain superior colliculus and deep mesencephalic nucleus. Other regions that contain less dense regions of combined anterograde and retrograde labeling include the following nuclei: the interstitial nucleus of medial longitudinal fasciculus, the pontine reticular formation, and the lateral periaqueductal gray. Premotoneurons that receive dense inputs from the wMCx are likely to be important mediators of cortical regulation of whisker movements and may be a key component in a central pattern generator involved in the generation of rhythmic whisking activity.     

C-fos Expression During Vocal Mobbing in the New World Monkey Saguinus fuscicollis

In order to find brain areas involved in the vocal expression of emotion, we compared c-fos expression in three groups of saddle-back tamarins (Saguinus fuscicollis). One group, consisting of three animals, was made to utter more than 800 mobbing calls by electrical stimulation of the periaqueductal grey of the midbrain (PAG). A second group, consisting of two animals, was stimulated in the PAG with the same intensity and for the same duration as the first group but at sites that did not produce vocalization. These sites lay somewhat medial to the vocalization-eliciting sites. A third group, consisting of two animals, was stimulated at vocalization-eliciting sites in the PAG but with an intensity below vocalization threshold. Fos-like immunoreactivity that was found in the vocalizing but not in the non-vocalizing animals was located in the dorsomedial and ventrolateral prefrontal cortex, anterior cingulate cortex, ventrolateral premotor cortex, sensorimotor face cortex, insula, inferior parietal cortex, superior temporal cortex, claustrum, entorhinal and parahippocampal cortex, basal amygdaloid nucleus, anterior and dorsomedial hypothalamus, nucleus reuniens, lateral habenula, Edinger-Westphal nucleus, ventral and dorsolateral midbrain tegmentum, nucleus cuneiformis, sagulum, pedunculopontine and laterodorsal tegmental nuclei, ventral raphe, periambigual reticular formation and solitary tract nucleus. For some of these structures (e.g. anterior cingulate cortex and periambigual reticular formation), there is evidence also from electrical stimulation, lesioning and single-unit recording studies that they are involved in vocal control. For other structures (e.g. lateral habenula, Edinger-Westphal nucleus), the available evidence speaks against such a role. Fos activation in these cases is probably related to non-vocal reactions accompanying the electrically elicited vocalizations. A third group of structures consists of areas for which a role in vocal control cannot be excluded but for which the present study presents the first evidence for such a role (e.g. claustrum and sagulum). These structures deserve further studies using more specific methods.     

Serotonergic projections to the caudal brain stem: a double label study using horseradish peroxidase and serotonin immunocytochemistry

Cells of origin of serotonergic and non-serotonergic projections to the caudal brain stem in the primate were examined using a double label technique. Following HRP injections into medullary raphe nuclei and the adjacent reticular formation double labeled cells were found in the dorsal raphe nucleus, the central superior nucleus and the ventrolateral tegmentum. Retrogradely labeled cells that did not stain for serotonin-like immunoreactivity were found primarily in the periaqueductal gray (PAG) and the mesencephalic and pontine reticular formation. The results are discussed in relation to the descending pathway(s) mediating the effects of PAG stimulation.     

Single unit recording in the midbrain of rats during shock-elicited fighting behavior

Single unit activity was recorded extracellularly from the midbrain of rats during fighting behavior and during non-fighting control manipulations. Fighting behavior was elicited by footshock or startle stimuli or occurred spontaneously as a result of prior footshock presentations.

Seven cells were found in the midbrain reticular formation and central gray which displayed maximum firing rates during fighting behavior. These cells also fired to a limited extent to some of the control manipulations, particularly contralateral vibrissae stimulation. These cells fired phasically during fighting behavior and their firing was correlated with either the approach or paw-strike of the opponent animal or to the response of the recording animal to a tactic of the opponent animal. However, no specific movement or sensory event reliably predicted the firing of these cells during fight sequences.

Cells located in other midbrain areas, such as the deep tectum or the area of the red nucleus, also responded during fighting behavior. However, the discharge of these cells was correlated with specific body movements or sensory events. The activity during fighting was similar in rate and pattern to activity during control manipulations whenever similar movements or sensory stimulation were produced. Cells were also found which did not discharge during fighting behavior although they fired under a variety of other conditions.     

Neuroanatomy of Cardiac Activity-regulating Circuitry: A Transneuronal Retrograde Viral Labelling Study in the Rat

The anatomy of cardiac activity-regulating circuitry was studied with retrograde transneuronal viral labelling after pseudorabies virus injections into different parts of the rat heart. Transection of the spinal cord at Th1 was used to reveal selectively the parasympathetic neuronal networks. Virus-labelled sympathetic preganglionic cells were found in the Th1-Th7 thoracic intermediolateral cell groups, with some additional infections at Th8-Thll after inoculations of the ventricular myocardium. After ventricular injections the thoracic spinal labelling pattern was bilateral and after right atrial infection it was contralateral. Approximately 20% of the parasympathetic preganglionic cells were located in the dorsal motor vagus nucleus; the rest occupied positions in the peri-ambiguus area ventrolateral to the nucleus ambiguus. Here and in the ventrolateral reticular formation myocardiotopy was found. Supraspinal transneuronal infections were bilateral, showed no apparent side dominance and were found in the nucleus of the solitary tract, the area postrema, the raphe nuclei, the A5 group, the parabrachial region, the periaqueductal grey, the hypothalamus, the amygdala and the cortex, in particular the anterior cingulate, the frontal, prelimbic, infralimbic and insular cortices. Spinal transections at Th1 reduced the number of labelled cells, gave a right side labelling dominance and affected the infection patterns in the ventrolateral reticular area, the raphe nuclei, the periaqueductal grey matter, the perifornical and retrochiasmatic area and the rostra1 parts of the insular cortex. The latter structures are linked selectively to the sympathetic innervation of the heart. The anatomical and functional aspects of these findings are discussed in relation to the autonomic control of heart activity.     

Connections of the corpus cerebelli in the thornback guitarfish, Platyrhinoidis triseriata (Elasmobranchii): a study with WGA-HRP and extracellular granule cell recording

The neuronal connections of the cerebellar corpus in the guitarfish Platyrhinoidis triseriata were investigated by WGA-HRP injections and extracellular recording of sensory evoked electrical activity. Injections of WGA-HRP into the corpus resulted in retrograde labeling of the following cell groups bilaterally: pretectal and accessory optic nuclei, interstitial nucleus of Cajal, nucleus ruber, oculomotor and possibly trochlear nucleus, central (periaqueductal) gray, nucleus H, reticular formation of the midbrain, cerebellar nucleus, caudal part of nucleus F, tentatively locus coeruleus and subcoeruleus field, octaval and trigeminal nuclei, intermediate octavolateralis nucleus, medial inferior reticular formation, lateral reticular nucleus, and spinal cord. Unilaterally labeled cells were seen in the contralateral inferior olive, which was found to project in sagittal zones onto the molecular layer of the corpus. Terminal fields of efferent Purkinje cell axons were labeled over the ipsilateral cerebellar nucleus exclusively. Purkinje cells in different parts of the corpus project topographically onto subdivisions of the nucleus. Mapping of evoked electrical multiple unit activity recorded from the granule cell layer of the corpus shows separate visual and tactile areas, mostly confined to the anterior and posterior lobes, respectively. Granule cells within the tactile area also responded to lateral line stimuli and, at two distinct medial locations in the caudal and rostral parts of the posterior lobe, to weak electric field stimulation in the bath. The body surface is somatotopically represented in the tactile area, but discontinuities in the map might indicate that the somatotopy is "fractured".     

Brain stem projections to the facial nucleus of the rat

Horseradish peroxidase was injected into the medial and lateral columns of the facial nucleus of the rat. Following medial injections, cells were labelled by retrograde transport in the ipsilateral spinal trigeminal nucleus (caudalis) both medial vestibular nuclei, contralateral midbrain reticular formation and nucleus of the lateral lemniscus. The periaqueductal grey, interstitial nucleus and nucleus of Darkschewitch were also labelled ipsilaterally. Injections into the lateral column of the facial nucleus labelled the spinal trigeminal nucleus (oralis) and parabrachial nuclei ipsilaterally and the Darkschewitch and red nuclei contralaterally.     

The effects of decerebration and destruction of nucleus raphe magnus, periaqueductal grey matter and brainstem lateral reticular formation on the depression due to surgical trauma of the jaw-opening reflex evoked by tooth-pulp stimulation in the cat

The effects on the jaw-opening reflex evoked by tooth-pulp stimulation of surgical trauma, decerebration and the destruction of a number of nuclei associated with descending inhibition of trigeminal or spinal neurones have been investigated in the cat. Surgical preparation caused a progressive elevation of the digastric reflex threshold. After decerebration, reflex thresholds remained elevated for 8–11 h before returning to close to pre-surgical control values. Destruction of the nucleus raphe magnus and of the periaqueductal grey matter did not affect the depressed reflex in decerebrate or anaesthetized cats. Variable effects were produced by bilateral ablation of the juxta-raphe reticular formation and destruction of the rostral ipsilateral lateral reticular formation of the brainstem.     

Afferents to the nucleus reticularis parvicellularis of the cat medulla oblongata: A tract-tracing study with cholera toxin B subunit

The aim of this study was to examine anatomical evidence in cats of whether the nucleus reticularis parvicellularis (Pc) is part of the circuit responsible for the inhibition of brainstem motoneurons during paradoxical sleep. For this purpose, we made iontophoretic injections of the retrograde and anterograde tracer cholera toxin B subunit (CTb) in the Pc. After CTb injections in the Pc, a large number of retrogradely labeled neurons were seen in the central nucleus of the amygdala, the lateral part of the bed nucleus of the stria terminalis, the posterior hypothalamic areas, the mesencephalic reticular formation, the nucleus locus subcoeruleus, the nucleus pontis caudalis, other portions of the Pc, the nucleus reticularis dorsalis, the trigeminal sensory complex, and the nucleus of the solitary tract. We further found that the Pc receives (1) serotoninergic afferents from the raphe dorsalis, magnus, and obscurus nuclei; (2) noradrenergic inputs from the dorsolateral pontine tegmentum; (3) cholinergic afferents from the lateral medullary reticular formation; (4) substance P-like afferents from the central nucleus of the amygdala, bed nucleus of the stria terminalis, periaqueductal gray, and nucleus of the solitary tract; and (5) methionine-enkephalin-like projections from the periaqueductal gray, the nucleus of the solitary tract, the lateral pontine and medullary reticular formation, and the spinal trigeminal nucleus. We further found that the Pc do not receive afferents from brainstem structures responsible for muscle atonia, such as the ventromedial medulla and the dorsomedial pontine tegmentum, and therefore may not be part of the circuit inhibiting the brainstem motoneurons during paradoxical sleep. © 1994 Wiley-Liss, Inc.     

Afferent connections of the thalamic intralaminar nuclei in the cat

Afferents to the central lateral (CL), paracentral (PC) and central medial (CE) intralaminar nuclei (ILN) from cortical and subcortical sites were studied in the cat. We utilized stereotaxically guided injections of HRP into the CL and PC nuclei and tritiated leucine injections into various visual, parietal and limbic areas of cortex to demonstrate these connections. In studying the relatively weak visual cortical projections to the ILN, we demonstrated projections from areas 19, 20a, 21a, 21b, AMLS, PMLS and PLLS. However, our HRP injections into the ILN often revealed only a few labeled cells in any of the above areas; therefore conclusions regarding the absence of projections to ILN from remaining visual cortical areas should be made cautiously. The ILN receive heavier projections from the frontal eye fields, cingulate cortex, splenial cortex, insular cortex, somatosensory areas SI and SII, auditory areas SF, AII, and Ep, and parietal areas 5 and 7. The most robust projections appear to be from from frontal eye fields, cingulate and parietal areas. No topography was apparent in the projections to the ILN. All cortical projections originate ipsilaterally from layers V and VI. Heavy subcortical projections to the ILN originate in the pretectum, superior colliculus, reticular formation, and periaqueductal grey. Fewer afferents arise from several other brainstem and thalamic nuclei.