Projections of the paleostriatum upon the midbrain tegmentum in the pigeon

Descending projections of neurons in the avian paleostriatal complex upon cell groups of the midbrain tegmentum were investigated in the pigeon using horseradish peroxidase histochemistry and autoradiography. The purpose of this study was to determine if pathways comparable to the mammalian striato-tegmental system (descending paths from neurons in the caudate nucleus, putamen and nucleus accumbens) exist in the avian brain. Although previous reports (Karten &Dubbeldam, 1973) have described pathways in the avian brain comparable to the mammalian pallido-tegmental system, no counterpart of the mammalian striato-tegmental system has been identified in birds.Our results indicate that a striato-tegmental system does exist in the pigeon. Neurons within a small-celled field containing a rich plexus of catecholamine-positive axons and terminals were found to project upon the midbrain tegmentum. This small-celled field includes the paleostriatum augmentatum, lobus parolfactorius and nucleus accumbens. Cells in all three components of this small-celled field project upon the pars compacta and the pars dorsomedialis of the nucleus tegmenti pedunculo pontinus of the midbrain. The pars compacta of this nucleus contains many catecholamine-positive cells and has been compared to the substantia nigra pars compacta of mammals on the basis of histochemistry and anatomical connections (Brauth,Ferguson &Kitt, 1978). The pars dorsomedialis of this nucleus has been shown to contain a plexus of substance P-positive axons and terminals (Reiner,Brauth,Kitt &Karten, 1980;Reiner,Karten &Korte, 1980). It is argued that the pars dorsomedialis of the pedunculo pontine nucleus of the pigeon is comparable to the mammalian substantia nigra pars reticulata. Other projection targets of the lobus parolfactorius and nucleus accumbens in the pigeon include the area ventralis of Tsai, the lateral habenular nucleus and the nucleus subceruleus dorsalis. The locus ceruleus also receives a projection from the nucleus accumbens. The results are compared to previously reported findings in mammals and reptiles.     

Projections of striatopallidal structures to the pedunculopontine nucleus of the tegmentum of the midbrain in dogs

A method based on retrograde axonal transport of horseradish peroxidase was used to study the striatopallidal afferent projections of the pendunculopontine nucleus of the midbrain (PPN) in dogs. The major source of these projections was found to be the pallidum, as both the compact and diffuse zones of this nucleus received projections from all of its structures: the entopeduncular nucleus, the globus pallidus, and the ventral pallidum. The striatal complex, specifically the nucleus accumbens, showed only occasional labeled neurons, projecting exclusively to the compact part of the PPN. Since the distribution of projection fibers arising from the functionally diverse territories of the striatopallidum and directed to individual structural subdivisions of the PPN showed no topical elements, identification of functionally specific (motor and limbic) areas of the PPN was not possible on the basis of the present analysis. __________ Translated from Morfologiya, Vol. 130, No. 6, pp. 30–34, November–December, 2006.     

Projections from the raphe nuclei to the suprachiasmatic nucleus of the rat

The presence of serotonergic afferents in the hypothalamic suprachiasmatic nucleus (SCN) is well documented and several functional roles of serotonin (5-HT) in circadian function are well established. However, there is some controversy about the precise location of the serotonergic neurones from where this input arises. Discrete injection of the tracer Cholera toxin, subunit B, (ChB) was centred in the rat SCN, and a few retrograde labelled neurones were distributed in the dorsal and median raphe nuclei (MnR) and in the rostral part of the raphe magnus (RMg), but no neurones were found in the raphe pallidus or raphe obscurus. In addition, a group of neurones was consistently found in the medial part of the pontine supra lemniscal nucleus but not including the serotonergic B₉ region. A combination of retrograde tracing with Fluoro-Gold together with 5-HT-immunolabelling, showed few double-labelled neurones in the dorsal, MnR and B₉. However, the majority of projecting neurones were not co-storing 5-HT immunoreactivity. Phaseolus vulgaris-leucoagglutinin (PHA-L) injections in the dorsal raphe resulted in faint labelling, whereas the MnR gave rise to several labelled fibres in the SCN. Individual delicate PHA-L nerve fibres were found in all compartments of the SCN both in terms of rostrocaudal, ventromedial and dorsomedial extent, without any sign of a topographical organisation of the MnR input to the SCN. PHA-L injections into RMg gave rise to labelling of a few processes within the SCN. In conclusion, the main serotonergic input to the rat SCN originates from a few neurones in the MnR.     

Analysis of behavioral deficits produced by lesions in the dorsal and ventral midbrain tegmentum

The impact of midbrain lesions on ingestive behavior in the rat was examined under a variety of pharmacological and dietary conditions. Electrolytic lesions in the dorsal midbrain tegmentum, ventrolateral to the central gray, produced moderate to severe (4 to 19 days) aphagia and adipsia in the rat. This effect was followed by complete recovery of ad lib food intake but a sustained suppression of water intake and urine output. Caloric regulation, drug-induced anorexia, drinking in response to intracellular and extracellular dehydration, and feeding as a function of dietary palatability appeared normal. Persistent deficits, however, were observed in the rats' feeding response to glucoprivation, in their consumption of sucrose or saccharin solutions, in post-fast compensatory food intake, and in their ability to maintain a nocturnal pattern of feeding. In contrast to these changes, electrolytic lesions in the ventral midbrain tegmentum produced an increase in food intake and body weight gain. This effect was associated with no other food- or water-related behavioral changes, with the exception of amphetamine- or mazindol-induced anorexia which was attenuated or abolished by the lesion.     

Projections to the midbrain from the medial versus lateral prefrontal cortices of the rat

Microinjections of horseradish peroxidase were made into the medial or lateral prefrontal cortex (PFC) of rats. The trajectory of descending, orthogradely labeled axons in route to the midbrain was then charted. The descending pathways originating from the medial versus lateral PFC were noticeably different. In general the medial PFC projected to dorsal aspects of the midbrain (e.g. dorsolateral periaqueductal gray and deep layers of superior colliculus), whereas the lateral PFC projected to the lateral and ventral aspects of the midbrain (e.g. ventral periaqueductal gray and laterally adjacent reticular formation). These differences suggest that the medial and lateral PFC may have different functions in regard to their potential influences upon the midbrain. Potential influences upon midbrain analgesic mechanisms are discussed.     

Projections from the rat pedunculopontine and laterodorsal tegmental nuclei to the anterior thalamus and ventral tegmental area arise from largely separate populations of neurons

Cholinergic and non-cholinergic neurons in the brainstem pedunculopontine (PPT) and laterodorsal tegmental (LDT) nuclei innervate diverse forebrain structures. The cholinergic neurons within these regions send heavy projections to thalamic nuclei and provide modulatory input as well to midbrain dopamine cells in the ventral tegmental area (VTA). Cholinergic PPT/LDT neurons are known to send collateralized projections to thalamic and non-thalamic targets, and previous studies have shown that many of the afferents to the VTA arise from neurons that also project to midline and intralaminar thalamic nuclei. However, whether cholinergic projections to the VTA and anterior thalamus (AT) are similarly collateralized is unknown. Ultrastructural work from our laboratory has demonstrated that cholinergic axon varicosities in these regions differ both morphologically and with respect to the expression and localization of the high-affinity choline transporter. We therefore hypothesized that the cholinergic innervation to these regions is provided by separate sets of PPT/LDT neurons. Dual retrograde tract-tracing from the AT and VTA indicated that only a small percentage of the total afferent population to either region showed evidence of providing collateralized input to the other target. Cholinergic and non-cholinergic cells displayed a similarly low percentage of collateralization. These results are contrasted to a control case in which retrograde labeling from the midline paratenial thalamic nucleus and the VTA resulted in higher percentages of cholinergic and non-cholinergic dual-tracer labeled cells. Our results indicate that functionally distinct limbic target regions receive primarily segregated signaling from PPT/LDT neurons.     

Projections from the superior colliculus to the ventral lateral geniculate nucleus in the hereditarily microphthalmic rat

Projections from the superior colliculus (SC) to the ventral lateral geniculate nucleus (LGNv) were studied in hereditarily microphthalmic and normal rats by means of wheatgerm agglutinin conjugated with horseradish peroxidase (WGA-HRP). Unilateral injection of a tracer into the LGNv in normal rats revealed WGA-HRP positive neurons on both sides of the SC. In the ipsilateral SC, most of the labeled neurons were distributed in the upper part of the stratum opticum (SO) and the lower part of the stratum griseum superficiale (SGS). A few labeled neurons were also found in the same layers of the contralateral SC. After unilateral injections of the tracer into the LGNv of microphthalmic rats, labeled neurons appeared in similar layers of the SC on both sides. However, the number of labeled neurons in the ipsilateral SC decreased to 30% of normal, whereas on the contralateral side these neurons were apparently more numerous than those in normal rats. The soma size of the labeled SC neurons in microphthalmia was not significantly different from normal. These results indicate fundamentally that tecto-LGNv projecting neurons exist in microphthalmic rats despite the fact that they lack optic nerve afferents. Furthermore, the present results, taken together with our previous results, indicate that the diminution in the number of tecto-LGNd neurons was severest (3%), the tecto-LGNv neurons less severe (30%) and the tecto-LP neurons least severe (50% of that of normal).     

Pyramidal input to the intracerebellar nuclei of the cat

In unanesthetized cats it has been found that pyramidal volleys elicited upon medullary pyramidal tract stimulation were capable of modifying the discharge of 41% of intracerebellar nuclear cells, via pontocerebellar systems impinging predominantly on the lateral cerebellar cortex. The incidence of responsive cells was 80% in the dentate nucleus compared with 10% in the fastigial nucleus, 11% in the anterior and 12% in the posterior division of the interpositus nucleus. The response was in 59% of the cases excitation followed by inhibition, in 30% of the cases a pure excitation and in 11% of the cases a pure inhibition. Excitation, pure or followed by inhibition, had a mean latency of 5.78 ms and a mean duration of 12.21 ms, while inhibition displayed a mean latency of 9.03 ms and a mean duration of 34.64 ms. The possible functional significance of the pyramidal input to the lateral cerebellum is briefly discussed in relation to a possible convergence of pyramidal and associative impulses in single cerebellar neurons.     

Projections from the ventral tegmental area and mesencephalic raphe to the dorsal raphe nucleus in the rat

Summary The origins of the dopaminergic innervation of the rat dorsal raphe nucleus (NRD) have been investigated using a combination of fluorescent retrograde tracing and fluorescence histochemistry. Stereotaxic microinjections of True Blue were placed in the central, caudal and lateral portions of the NRD, and after 6–12 days survival the brains were processed for fluorescence histochemical detection of catecholamines. Retrogradely labeled neurons were searched for in the diencephalic A11 and A13 dopaminergic cell groups, substantia nigra, ventral tegmental area (VTA) and the linear, central superior and dorsal raphe nuclei. The various NRD injections consistently resulted in retrograde labeling of a small number of catecholamine-containing, presumed dopaminergic cell bodies, confined mainly to three regions: the VTA, the linear and central superior raphe nuclei and the NRD itself. The present findings indicate that not only dopaminergic neurons in the VTA but also the system of catecholamine-containing cells, extending dorsally and caudally from the VTA within the midline raphe area, project to the NRD. Although often similar in size, shape and distribution to the catecholaminergic neurons the majority of retrogradely labeled cells in these regions were, however, found to be non-catecholaminergic.Abbreviations 3 Principal oculomotor nucleus - 4 Trochlear nucleus - Aq Cerebral aqueduct - cp cerebral peduncle - cst cortico-spinal tract - dscp decussation of the superior cerebellar peduncle - DTg Dorsal tegmental nucleus - fr fasciculus retroflexus - IF Interfascicular nucleus - IP Interpeduncular nucleus - LL nucleus of the lateral lemniscus - ml medial lemniscus - mlf medial longitudinal fasciculus - mNV mesencephalic trigeminal nucleus - NLC Nucleus linearis caudalis - NLR Nucleus linearis rostralis - NRD Dorsal raphe nucleus - PAG Periaqueductal grey - PN Pontine nucleus - PRN Pontine raphe nucleus - R Red nucleus - RCS Nucleus raphe centralis superior - SN Substantia nigra - VTA Ventral tegmental area - VTg Ventral tegmental nucleus     

Topography and functional role of dopaminergic projections from the ventral mesencephalic tegmentum to the ventral pallidum.

A dopaminergic projection from the ventral tegmental area to the ventral pallidum was identified in the rat using anterograde tract tracing and combined retrograde tracing-immunocytochemistry. The projection was found to be topographically organized such that fibers innervating the ventromedial ventral pallidum arose from neurons located along the midline nuclei of the ventral mesencephalon, including the nucleus interfascicularis and nucleus linearis caudalis. Ventral tegmental neurons situated more laterally, in the nucleus parabrachialis pigmentosus and nucleus paranigralis, projected to the ventromedial and dorsolateral ventral pallidum. The substantia nigra did not supply a major contribution to this projection. The proportion of ventral tegmental area dopaminergic neurons projecting to the ventral pallidum ranged from approximately 30% to 60%. The functional significance of the projection is indicated since intra-ventral pallidum microinjections of dopamine elicited a dose-dependent increase in locomotor activity. Furthermore, whereas pretreatment of the ventral pallidum with the GABAA agonist muscimol has been shown to attenuate opioid-induced locomotor activity elicited from the ventral pallidum, it did not attenuate the dopamine-induced motor response. Thus, while mu-opioids in the ventral pallidum may presynaptically regulate GABAergic efferents from the nucleus accumbens, it appears that the dopaminergic input directly influences the ventral pallidal output neuron which is involved in locomotion.     

Metabolic activity of intracerebellar nuclei in the rat: Effects of inferior olive inactivation

Summary Metabolic activity of the intracerebellar nuclei during cryoinactivation of the inferior olive was studied in the anaesthetized rat by using the ¹⁴C-2-deoxyglucose method. Single unit recording of Purkinje cells was simultaneously monitored in the cerebellar cortex.Local inactivation in the inferior olive resulted in regional suppression of complex spike discharges in the cerebellar cortex.An increased metabolic activity was observed in the cerebellar nuclei contralateral to the cryoinactivation site correlating the somatotopically arranged olivo-cerebello-nuclear circuit. This increase was shown to be due specifically to inactivation of the inferior olive, since it was not obtained in a rat in which the inferior olive was previously destroyed by neurotoxic doses of 3-acetylpyridine.The results are interpreted as being due to an increased presynaptic activity of the terminals of the Purkinje cells which fire simple spikes at high rates after climbing fibre deafferentation.     

Projections from auditory cortex to midbrain cholinergic neurons that project to the inferior colliculus

We have shown that auditory cortex projects to cholinergic cells in the pedunculopontine tegmental nucleus (PPT) and laterodorsal tegmental nucleus (LDT). PPT and LDT are the sources of cholinergic projections to the inferior colliculus, but it is not known if the cortical inputs contact the cholinergic cells that project to the inferior colliculus. We injected FluoroRuby into auditory cortex in pigmented guinea pigs to label cortical projections to PPT and LDT. In the same animals, we injected Fast Blue into the left or right inferior colliculus to label PPT and LDT cells that project to the inferior colliculus. We processed the brain to identify cholinergic cells with an antibody to choline acetyltransferase, which was visualized with a green fluorescent marker distinguishable from both FluoroRuby and Fast Blue. We then examined the PPT and LDT to determine whether boutons of FluoroRuby-labeled cortical axons were in close contact with cells that were double-labeled with the retrograde tracer and the immunolabel. Apparent contacts were observed ipsilateral and, less often, contralateral to the injected cortex. On both sides, the contacts were more numerous in PPT than in LDT. The results indicate that auditory cortex projects directly to brainstem cholinergic cells that innervate the ipsilateral or contralateral inferior colliculus. This suggests that cortical projections could elicit cholinergic effects on both sides of the auditory midbrain.     

Projections from the raphe nuclei to the phrenic motor nucleus in the cat

The monoclonal antibody HNK-1 binds to a carbohydrate determinant in the myelin-associated glycoprotein (MAG) and other glycoproteins of human peripheral nerve. Some glycoproteins of lower Mr than the major P0 glycoprotein of myelin appear to bind more antibody than MAG. These glycoproteins electrophorese in the Mr range of 20,000 to 26,000 and are present in the purified myelin fraction. The results indicate that an antigen on the surface of a subset of lymphocytes is shared with a group of glycoproteins in human peripheral nerve. The antigen appears to be similar to that recognized by IgM paraproteins associated with a type of neuropathy.     

Occurrence of dopamine-containing neurons in the midbrain raphe nuclei of the rat

Formaldehyde-induced fluorescence histochemistry, by the use of the improved filter system, revealed that dorsal and median raphe nuclei contain in varying numbers blue-green fluorescent neurons among a large number of yellow fluorescent serotonin-containing neurons. Pharmacological treatments indicated the presence of catecholamine in the blue-green fluorescent neurons. Moreover, microspectrofluorometry identified the catecholamine therein as dopamine, based on an excitation maximum shift characteristic of dopamine fluorescence caused by HCl vapor. Significance of these dopaminergic neurons in the raphe nuclei is suggested.     

Are there serotonergic projections from raphe and retrotrapezoid nuclei to the ventral respiratory group in the rat?

The relationship of serotonergic neurons in raphe and ventrolateral medullary regions to neurons projecting to the rostral ventral respiratory group in the rat were investigated using combined immunohistochemical and retrograde labeling techniques. Serotonergic and non-serotonergic neurons retrogradely labeled with rhodamine beads were found intermingled in the larger population of serotonin-immunoreactive neurons in raphe pallidus, obscurus and magnus. In addition, a cell group analogous to the retrotrapezoid nucleus in the cat was identified in the rat ventrolateral medulla. Retrotrapezoid neurons, which exhibited exclusively ipsilateral projections to the ventral respiratory group, were located lateral to clusters of serotonergic cells near the ventral surface, but were not serotonin immunoreactive.     

Inhibitory control of intracerebellar nuclei by the Purkinje cell axons

Summary In anaesthetized cats, synaptic events in cerebellar nuclei neurones were investigated with intracellular microelectrode techniques. These cells were identified by their antidromic activation along their axons and/or by their location in histological sections. In the cells of lateral nucleus IPSPs were induced monosynaptically during stimulation of the overlying hemispheral cortex of the cerebellum. In the cells of nuclei interpositus and fastigii, similar IPSPs were produced from the paravermal and vermal cortices, respectively. The postulate that the Purkinje cells exert an inhibitory action upon their target neurones thus applies not only to Deiters neurones, as previously proposed, but also to cells in the cerebellar nuclei. Stimulation of the cerebellar afferents at the inferior olive, the pontine nucleus and the lateral reticular nucleus produced EPSPs in cerebellar nuclei cells with relatively brief latencies, probably through axon collaterals of these afferents. The EPSPs were followed by IPSPs and slow depolarizations of disinhibitory nature, which, as studied previously in Deiters neurones, might be caused respectively by activation and subsequent depression of Purkinje cells through the cerebellar intracortical mechanisms.     

Projections from auditory structures to the superior colliculus in the rat

Arginine-vasopressin (AVP) and related peptides were administered by microinjection into the nucleus tractus solitarius of the rat. AVP produced a rise both in mean arterial pressure and in heart rate. This effect was abolished by pretreatment with a specific antagonist of V1 receptors and was not seen either after injections of oxytocin or of the V2 agonist deamino-D-arginine-vasopressin. This study provides evidence for a specific action of vasopressin on the cardiovascular system in the nucleus of the tractus solitarius, which is mediated neither by V2 nor oxytocin receptors.