Projections of cholinergic and non-cholinergic neurons of the brainstem core to relay and associational thalamic nuclei in the cat and macaque monkey

The projections of brainstem core neurons to relay and associational thalamic nuclei were studied in the cat and macaque monkey by combining the retrograde transport of wheat germ agglutinin conjugated with horseradish peroxidase with choline acetyltransferase immunohistochemistry. All major sensory (medial geniculate, lateral geniculate, ventrobasal), motor (ventroanterior, ventrolateral, ventromedial), associational (mediodorsal, pulvinar, lateral posterior) and limbic (anteromedial, anteroventral) thalamic nuclei of the cat were found to receive projections from cholinergic neurons located in the peribrachial area of the pedunculopontine nucleus and in the laterodorsal tegmental nucleus as well as from non-cholinergic neurons in the rostral (perirubral) part of the central tegmental mesencephalic field. Specific relay nuclei receive less than 10% of their brainstem afferents from non-cholinergic neurons located at rostral midbrain levels and receive 85-96% of their brainstem innervation from a region at midbrain-pontine junction where the cholinergic peribrachial area and laterodorsal tegmental nucleus are maximally developed. Of the total number of horseradish peroxidase-positive brainstem neurons seen after injections in various specific relay nuclei, the double-labeled (horseradish peroxidase + choline acetyltransferase) neurons represent approximately 70-85%. Three to eight times more numerous horseradish peroxidase-labeled brainstem cells were found after injections in associational (mediodorsal and pulvinar-lateral posterior complex) and diffusely cortically-projecting (ventromedial) thalamic nuclei of cat than after injections in specific relay nuclei. The striking retrograde cell labeling observed after injections in nuclei with associative functions and widespread cortical projections was due to massive afferentation from non-cholinergic parts of the midbrain and pontine reticular formation, on both ipsi- and contralateral sides. After wheat germ agglutinin-horseradish peroxidase injections in the associative pulvinar-lateral posterior complex and mediodorsal nucleus of Macaca sylvana, 45-50% of horseradish peroxidase-positive brainstem peribrachial neurons were also choline acetyltransferase-positive. While cells in the medial part of the cholinergic peribrachial area were found to project especially towards the pulvinar-lateral posterior nuclear complex in monkey, the retrograde cell labeling seen after the mediodorsal injection was mostly confined to the lateral part of both dorsal and ventral aspects of the peribrachial area.(ABSTRACT TRUNCATED AT 400 WORDS)     

Cholinergic and non-cholinergic projections from the upper brainstem core to the visual thalamus in the cat

Summary The projections of cholinergic and noncholinergic neurons of the rostral brainstem reticular formation to the visual thalamic nuclei (dorsal lateral geniculate — LG, lateral posterior — LP, and perigeniculate — PG) were studied in cat by using the retrograde transport of horseradish peroxidase conjugated with wheat germ agglutinin (WGA-HRP) combined with choline acetyltransferase (ChAT) immunohistochemistry. After thalamic injections, less than 10% of all retrogradely labeled neurons in the upper brainstem reticular core were located at most rostral (perirubral) levels where there are virtually no cholinergic elements. Approximately 75–80% of all HRP-positive neurons in the reticular formation were found between stereotaxic planes anterior 1 and posterior 2, in the peribrachial (PB) area of the pedunculopontine nucleus and in the laterodorsal tegmental (LDT) nucleus. The brainstem afferents to LG and PG thalamic nuclei essentially derive from PB neurons, with a small contribution from LDT cells, whereas the LP thalamic nucleus receives massive inputs from both PB and LDT brainstem nuclei. Of all HRP-positive elements visualized in the PB nucleus after an LG or a PG injection, 87% and 73%, respectively, were also ChAT-positive. Of all HRP-positive elements in the PB and LDT nuclei after an LP injection, 82% and 92%, respectively, were also ChAT-positive. The numbers of labeled neurons in the contralateral brainstem reticular nuclei reach 30% to 50% of the numbers found in the ipsilateral reticular formation. These findings reveal the existence of a prominent cholinergic projection from the brainstem reticular formation to the visual thalamic nuclei. Such a chemospecific projection is probably involved in phasic and tonic events of activated behavioral states.After this paper was accepted for publication,we read a study by DeLima and Singer (J Comp Neurol 259:92-121, 1987) on cholinergic and monoaminergic afferents to the LG nucleus. The results reported in that study fit in well with our data.     

Connections of the zona incerta to the reticular nucleus of the thalamus in the rat

This study demonstrated that there is a pathway from the zona incerta to the thalamic reticular nucleus. Injections of horseradish peroxidase or Fluorogold were made, using stereotaxic coordinates, into the rostral, intermediate or caudal regions of the thalamic reticular nucleus of adult Sprague-Dawley rats. The results show that the different regions of the thalamic reticular nucleus have distinct patterns of connections with the sectors of the zona incerta. In terms of the relative strength of the connections, injections made into the rostral regions of the thalamic reticular nucleus showed the highest number of labelled cells within the rostral and ventral sectors of the zona incerta; injections made into the intermediate regions of the thalamic reticular nucleus showed labelled cells in the dorsal and ventral sectors; while injections to the caudal regions of the thalamic reticular nucleus showed only a few labelled cells in the caudal sector of the zona incerta. Previous studies have shown that the zona incerta projects to the higher order thalamic nuclei but not first order thalamic nuclei. The labelling observed in the present study may represent collaterals of zona incerta to higher order thalamic nuclei projections.     

Cholinergic and non-cholinergic projections from the canine pontomesencephalic tegmentum (Ch5 area) to the caudal intralaminar thalamic nuclei

Summary The distribution and morphology of cholinergic and non-cholinergic neurons projecting to the caudal intralaminar thalamic nuclei from the Ch5 area in the dog were examined using a technique combining horseradish peroxidase (HRP) retrograde labeling with choline acetyltransferase (ChAT) immunocytochemistry. After processing for ChAT, cholinergic neurons were found primarily within the nucleus tegmenti pedunculopontinus (PPN) and the central tegmental tract (ctt). ChAT positive neurons were also located in the nucleus cuneiformis and among the fibers of the lateral lemniscus and medial longitudinal fasciculus. On the basis of immunocytochemical and cytoarchitectonic data, PPN was divided into two distinct cell groups — a compact cell group located dorsolateral to the brachium conjunctivum and a diffuse cell group intermingled among the fibers of the brachium conjunctivum. Tissue processed for WGA-HRP and ChAT following injections of lectin-conjugated horseradish peroxidase into either the centrum medianum (CM) or parafascicular (Pf) nucleus resulted in double labeled cholinergic projection neurons in both PPN and ctt. Injections which involved CM and the caudal part of the central lateral thalamic nucleus (CL) resulted in more retrogradely labeled neurons than did those injections involving Pf. Injections of CM and CL also resulted in more double labeled cells in the dorsolateral compact portion of PPN than did injections confined to Pf. In all cases a small number of cholinergic neurons located in the contralateral PPN were retrogradely labeled as well. A substantial number of retrogradely labeled neurons were not ChAT positive, and in some cases, comprised up to 27% of the total population of projection neurons. Measurements of cell soma areas indicated that cells comprising the general cholinergic population were mostly medium (300–600 m²) or large (>600 m²) in size. The majority of cholinergic projection neurons fell within the medium size category while the non-cholinergic projection neurons were significantly smaller than their cholinergic counterparts. The results of this study suggest that in the dog, Ch5 cholinergic neurons which project to the caudal intralaminar thalamic nuclei are medium in size and are located primarily within PPN and ctt. In addition, a parallel projection to the caudal intralaminar nuclei exists which originates from smaller, non-cholinergic neurons in these same regions. Based on the results of this study, it appears that cholinergic projections to intralaminar thalamic nuclei which in turn project to the neostriatum may be one of the pathways over which PPN can affect basal ganglia activity.     

An autoradiographic analysis of ascending projections from the medullary reticular formation in the rat

Ascending projections from the several nuclei of the medullary reticular formation were examined using the autoradiographic method. The majority of fibers labeled after injections of [3H]leucine into nucleus gigantocellularis ascended within Forel's tractus fasciculorum tegmenti which is located ventrolateral to the medial longitudinal fasciculus. Nucleus gigantocellularis injections produced heavy labeling in the pontomesencephalic reticular formation, the intermediate layers of the superior colliculus, the pontine and midbrain central gray, the anterior pretectal nucleus, the ventral midbrain tegmentum including the retrorubral area, the centromedian-parafascicular complex, the fields of Forel/zona incerta, the rostral intralaminar nuclei and the lateral hypothalamic area. Nucleus gigantocellularis projections to the rostral forebrain were sparse. Labeled fibers from nucleus reticularis ventralis, like those from nucleus gigantocellularis, ascended largely in the tracts of Forel and distributed to the pontomedullary reticular core, the facial and trigeminal motor nuclei, the pontine nuclei and the dorsolateral pontine tegmentum including the locus coeruleus and the parabrachial complex. Although projections from nucleus reticularis ventralis diminished significantly rostral to the pons, labeling was still demonstrable in several mesodiencephalic nuclei including the cuneiform-pedunculopontine area, the mesencephalic gray, the superior colliculus, the anterior pretectal nucleus, the zona incerta and the paraventricular and intralaminar thalamic nuclei. The main bundle of fibers labeled by nucleus gigantocellularis-pars alpha injections ascended ventromedially through the brainstem, just dorsal to the pyramidal tracts, and joined Forel's tegmental tract in the midbrain. With the brainstem, labeled fibers distributed to the pontomedullary reticular formation, the locus coeruleus, the raphe pontis, the pontine nuclei, and the dorsolateral tegmental nucleus and adjacent regions of the pontine gray. At mesodiencephalic levels, labeling was present in the rostral raphe nuclei (dorsal, median and linearis), the mesencephalic gray, the deep and intermediate layers of the superior colliculus, the medial and anterior pretectal nuclei, the ventral tegmental area, zona incerta as well as the mediodorsal and reticular nuclei of the thalamus. Injections of the parvocellular reticular nucleus labeled axons which coursed through the lateral medullary tegmentum to heavily innervate lateral regions of the medullary and caudal pontine reticular formation, cranial motor nuclei (hypoglossal, facial and trigeminal) and the parabrachial complex.(ABSTRACT TRUNCATED AT 400 WORDS)     

Thalamic distribution of zinc-rich terminal fields and neurons of origin in the rat

Several cortico-cortical and limbic-related circuits are enriched in zinc, which is considered as an important modulator of glutamatergic transmission. While heavy metals have been detected in the thalamus, the specific presence of zinc has not been examined in this region. We have used two highly sensitive variations of the Timm method to study the zinc-rich innervation in the rat thalamus, which was compared to the distribution of acetylcholinesterase activity. The origin of some of these zinc-rich projections was also investigated by means of retrograde transport after intracerebral infusions of sodium selenium (Na2SeO3). The overall zinc staining in the thalamus was much lower than in the neocortex, striatum or basal forebrain; however, densely stained terminal fields were observed in the dorsal tip of the reticular thalamic nucleus, the anterodorsal and lateral dorsal thalamic nuclei and the zona incerta. In addition, moderately stained zinc-rich terminal fields were found in the rostral intralaminar nuclei, nucleus reuniens and lateral habenula. Intracerebral infusions of Na2SeO3 in the lateral dorsal nucleus resulted in retrogradely labeled neurons that were located in the postsubiculum, and also in the pre- and parasubiculum. These results are the first to establish the existence of a zinc-rich subicular-thalamic projection. Similar infusions in either the intralaminar nuclei or the zona incerta resulted in labeling of neurons in several brainstem structures related to the reticular formation. Our results provide morphological evidence for zinc modulation of glutamatergic inputs to highly selective thalamic nuclei, arising differentially from either cortical limbic areas or from brainstem ascending activation systems.     

The organization of preoptic-medullary circuits in the male rat: evidence for interconnectivity of neural structures involved in reproductive behavior, antinociception and cardiovascular regulation.

The present studies used anatomical tract-tracing techniques to delineate the organization of pathways linking the medial preoptic area and the ventral medulla, two key regions involved in neuroendocrine, autonomic and sensory regulation. Wheatgerm agglutinin-horseradish peroxidase injections into the ventromedial medulla retrogradely labeled a large number of neurons in the medial preoptic area, including both the median and medial preoptic nuclei. The termination pattern of preoptic projections to the medulla was mapped using the anterograde tracers Phaseolus vulgaris leucoagglutinin and biotinylated dextran amine. Tracer injections into the preoptic area produced a dense plexus of labeled fibers and terminals in the ventromedial and ventrolateral pons and medulla. Within the caudal pons/rostral medulla, medial preoptic projections terminated heavily in the nucleus raphe magnus; strong anterograde labeling was also present in the pontine reticular field. At mid-medullary levels, labeled fibers focally targeted the nucleus paragigantocellularis, in addition to the heavy fiber labeling present in the midline raphe nuclei. By contrast, very little labeling was observed in the caudal third of the medulla. Experiments were also conducted to map the distribution of ventral pontine and medullary neurons that project to the medial preoptic area. Wheatgerm agglutinin-horseradish peroxidase injections in the preoptic area retrogradely labeled a significant population of neurons in the ventromedial and ventrolateral medulla. Ascending projections from the medulla to the preoptic area were organized along rostral-caudal, medial-lateral gradients. In the caudal pons/rostral medulla, retrogradely labeled cells were aggregated along the midline raphe nuclei; no retrograde labeling was present laterally at this level. By contrast, in the caudal half of the medulla, cells retrogradely labeled from the medial preoptic area were concentrated as a discrete zone dorsal to the lateral reticular nucleus; labeled cells were not present in the ventromedial medulla at this level. The present findings suggest that the medial preoptic area and ventral midline raphe nuclei share reciprocal connections that are organized in a highly symmetrical fashion. By contrast, preoptic-lateral medullary pathways are not reciprocal. These preoptic-brainstem circuits may participate in antinociceptive, autonomic and reproductive behaviors.     

猫丘脑中央外侧核的皮质下传入联系

本实验用HRP逆行性轴浆运输技术,对猫丘脑中央外侧核的传入纤维联系及其局部定位关系进行了观察。投射至丘脑中央外侧核尾侧区的主要核团包括:外侧膝状体腹核背侧带、丘脑网状核特别是它的背侧部、上丘深层,以同侧为主。板内核、丘脑下部外侧区和黑质网状部神经元的轴突终止在同侧丘脑中央外侧核吻侧区。丘脑中央外侧核全长的传入起自脑干网状结构和前庭神经核,呈双侧投射。前者以同侧为主,后者以对侧占优势。同侧未定带,顶盖前区、动眼神经核周围的细胞群、对侧三叉神经感觉主核、楔束核、薄束核以及小脑齿状核内也含有少量标记细胞。我们还观察到HRP注射中心区位于中央外侧核并扩散至丘脑腹前核者,同侧脚内核含大量HRP阳性细胞,而Gudden被盖腹侧核内充满密集的标记终末。这些结果表明,丘脑中央外侧核可能涉及多种感觉和运动功能。     

Glutamate immunoreactivity in the rat basilar pons: light and electron microscopy reveals labeled boutons and cells of origin of afferent projections.

Immunohistochemical methods that employed a polyclonal antiserum directed against a glutamate-hemocyanin conjugate were utilized to examine the rat basilar pontine nuclei at both light and electron microscopic levels in order to identify putative glutamatergic neural elements. A large number of cells ranging in size from 11 to 32 microns in diameter and present in all subdivisions and at all rostrocaudal levels of the basilar pons exhibited intense glutamate immunoreactivity. Immunoreactive punctate structures, confirmed by electron microscopy to be axon terminals, were homogeneously distributed throughout the pontine neuropil, although a somewhat greater accumulation was apparent medially at mid-levels of the basilar pons and laterally at more caudal levels. Immunolabeled axons were also present throughout the pontine nuclei. In order to demonstrate possible extrinsic sources of glutamate-immunoreactive axon terminals within the pontine gray, injections of wheat germ agglutinin-horseradish peroxidase were made directly into the basilar pons. Tissue was then evaluated for the presence of retrogradely transported wheat germ agglutinin-horseradish peroxidase and the same tissue sections processed for glutamate immunocytochemistry. Following this combined protocol, neuronal somata exhibiting both wheat germ agglutinin-horseradish peroxidase and glutamate immunoperoxidase reaction products were observed within layer Vb of the cerebral cortex, zona incerta, the dentate nucleus of the cerebellum, nucleus paragigantocellularis of the medullary reticular formation, and the dorsal column nuclei. Such double-labeled cells were considered to represent glutamatergic neurons that provide axonal projections to the basilar pons. Ultrastructural studies of the pontine nuclei confirmed the presence of glutamate immunogold labeling in dendrites, neuronal somata, axons, and axon terminals. Immunoreactive boutons contained round vesicles and primarily formed asymmetric synapses at various postsynaptic loci which included glutamate-immunolabeled dendritic profiles and somata. These results suggest that glutamatergic basilar pontine neurons form one segment of a multisynaptic pathway involving glutamatergic afferents to the basilar pons, glutamatergic pontocerebellar projection neurons, and the glutamatergic granule cells of the cerebellar cortex.     

Light and electron microscopic demonstration of hypothalamic projections to the parabrachial nuclei in the cat

Hypothalamic connections with the parabrachial nuclei in the cat were studied at light and electron microscopic levels following wheat germ agglutinin-horseradish peroxidase injections into the parabrachial nuclei and electrolytic lesions in the hypothalamus. The greatest concentration of retrogradely labeled neurons occurred in the paraventricular nucleus. Labeled neurons were also seen within the preoptic, anterior, lateral, dorsomedial and ventromedial hypothalamic nuclei. Hypothalamic lesions resulted in the degeneration of terminals forming axosomatic and axodendritic synapses in the parabrachial nuclei, particularly its lateral division. These findings support the idea that hypothalamic connections to specific regions of the parabrachial nuclei may underlie the topographical functional organization demonstrated for these brainstem nuclei.     

The cerebellar projection from the reticular formation of the brain stem in the rabbit

Summary By retrograde transport of horseradish peroxidase the reticulocerebellar projections were examined in twenty-six rabbits.After injections in the cerebellum retrogradely labeled neurons were more numerous in the caudal reticular formation (ventral and gigantocellular reticular nuclei) than in its rostral part (caudal and oral pontine reticular nuclei). The labeled cells were of all sizes, large, medium-sized and small. Giant cells were labeled only after injections in the posterior lobe vermis.After injections in the anterior lobe, the posterior vermis, the fastigial nucleus and the flocculus, retrogradely labeled neurons were found bilaterally in the ventral reticular nucleus, the gigantocellular reticular nucleus and the caudal pontine reticular nucleus. Some cases with posterior vermal and fastigial injections in addition showed labeled neurons bilaterally in the oral pontine reticular nucleus. There were no major side differences. The cases with injections in the anterior part of the paramedian lobule gave rise to only a few labeled cells in the gigantocellular reticular nucleus.Negative findings were consistently made in the mesencephalic reticular formation.     

Afferent connections of the zona incerta: a horseradish peroxidase study in the rat

Restricted microelectrophoretic injections either of free horseradish peroxidase or of horseradish peroxidase conjugated with wheat germ agglutinin were given to albino rats in order to study the afferent connections of structures of the subthalamic region. The results suggest that the zona incerta receives its main input from several territories of the cerebral cortex, the mesencephalic reticular formation, deep cerebellar nuclei, regions of the sensory trigeminal nuclear complex and the dorsal column nuclei. Substantial input to the zona incerta appears to come from the superior colliculus, the anterior pretectal nucleus and the periaqueductal gray substance, whereas many other structures, among which hypothalamic nuclei, the locus coeruleus, the raphe complex, the parabrachial area and medial districts of the pontomedullary reticular formation, seem to represent relatively modest but consistent additional input sources. The afferentation of neurons in Forel's fields H1 and H2 appears to conform to the general pattern outlined above. As pointed out in the Discussion, the present results provide hodological support for the classic concept according to which the zona incerta can be regarded as a rostral extent of the midbrain reticular core. Some of the possible physiological correlates of the fiber connections of the zona incerta in the context of the sleep-waking cycle, ingestive behaviors, somatic motor mechanisms, visual functions and nociceptive behavior are briefly discussed.     

A lectin horseradish peroxidase study of the origin of ascending fibers in the medial forebrain bundle of the rat. The lower brainstem

The origins of projections within the medial forebrain bundle from the lower brainstem were examined with the horseradish peroxidase technique. Labeled cells were found in at least 15 lower brainstem nuclei following injections of a conjugate or horseradish peroxidase and wheat germ agglutinin at various levels of the medial forebrain bundle. Dense labeling was observed in the following cell groups (from caudal to rostral): A1 (above the lateral reticular nucleus); A2 (mainly within the nucleus of the solitary tract); a distinct group of cell trailing ventrolaterally from the medial longitudinal fasciculus at the level of the rostral pole of the inferior olive; raphe magnus; nucleus incertus; dorsolateral tegmental nucleus (of Castaldi); locus coeruleus; nucleus subcoeruleus; caudal part of the dorsal (lateral) parabrachial nucleus; and raphe pontis. Distinct but light labeling was seen in raphe pallidus and obscurus, nucleus prepositus hypoglossi, nucleus gigantocellularis pars ventralis, and the ventral (medial) parabrachial nucleus. Sparse labeling was observed throughout the medullary and caudal pontine reticular formation. Several lower brainstem nuclei were found to send strong projections along the medial forebrain bundle to very anterior levels of the forebrain. They were: A1, A2, raphe magnus (rostral part), nucleus incertus, dorsolateral tegmental nucleus, raphe pontis and locus coeruleus. With the exception of the locus coeruleus, attention has only recently been directed to the ascending projections of most of the nuclei mentioned above. Evidence was reviewed indicating that fibers from lower brainstem nuclei with ascending medial forebrain bundle projections distribute to widespread regions of the forebrain.It is concluded from the present findings that several medullary cell groups are capable of exerting a direct effect on the forebrain and that the medial forebrain bundle is the major ascending link between the lower brainstem and the forebrain.     

A quantitative study of the brainstem cholinergic projections to the ventral part of the oral pontine reticular nucleus (REM sleep induction site) in the cat

The ventral part of the cat oral pontine reticular nucleus (vRPO) is the site in which microinjections of small dose and volume of cholinergic agonists produce long-lasting rapid eye movement sleep with short latency. The present study determined the precise location and proportions of the cholinergic brainstem neuronal population that projects to the vRPO using a double-labeling method that combines the neuronal tracer horseradish peroxidase–wheat germ agglutinin with choline acetyltransferase immunocytochemistry in cats. Our results show that 88.9% of the double-labeled neurons in the brainstem were located, noticeably bilaterally, in the cholinergic structures of the pontine tegmentum. These neurons occupied not only the pedunculopontine and laterodorsal tegmental nuclei, which have been described to project to other pontine tegmentum structures, but also the locus ceruleus complex principally the locus ceruleus and peri-, and the parabrachial nuclei. Most double-labeled neurons were found in the pedunculopontine tegmental nucleus and locus ceruleus complex and, much less abundantly, in the laterodorsal tegmental nucleus and the parabrachial nuclei. The proportions of these neurons among all choline acetyltransferase positive neurons within each structure were highest in the locus ceruleus complex, followed in descending order by the pedunculopontine and laterodorsal tegmental nuclei and then, the parabrachial nuclei. The remaining 11.1% of double-labeled neurons were found bilaterally in other cholinergic brainstem structures: around the oculomotor, facial and masticatory nuclei, the caudal pontine tegmentum and the praepositus hypoglossi nucleus. The disperse origins of the cholinergic neurons projecting to the vRPO, in addition to the abundant noncholinergic afferents to this nucleus may indicate that cholinergic stimulation is not the only or even the most decisive event in the generation of REM sleep.     

A cervical primary afferent input to vestibular nuclei as demonstrated by retrograde transport of wheat germ agglutinin-horseradish peroxidase in the rat.

Wheat germ agglutinin-horseradish peroxidase conjugate (WGA-HRP) was microiontophoretically injected into the vestibular nuclear complex of the rat. Retrogradely labeled neurons were found in ipsilateral spinal ganglia C2-C3 only if the injection site was in the caudal part of the medial vestibular nucleus (MVN). Injections into rostral parts of the MVN, the superior, lateral and descending vestibular nuclei (SVN, LVN, DVN), the nucleus of the solitary tract (STN) and the reticular formation did not result in spinal ganglion labeling. Thus, the caudal part of the MVN appears to be the main vestibular termination site for rostral cervical primary afferents.     

Cortical projections from the suprasylvian gyrus to the reticular thalamic nucleus in the cat

The cat's suprasylvian gyrus was injected iontophoretically with either 4% wheat germ agglutinin-horseradish peroxidase, 4% dextran-fluororuby or 4% dextran-biotin. The locations of labelled fibres, presumed terminals and cell bodies were determined with the aid of a camera lucida attachment and computer aided stereometry. Cells from the crown of the suprasylvian gyrus project to the dorsal-most portion of the rostral half of the reticular nucleus. The region or 'sector' is distinct, albeit with some overlap, from the visual sector of the reticular nucleus defined by projections from adjacent extrastriate visual cortices. The projection from the suprasylvian gyrus to the reticular nucleus has a rough topography such that the caudal areas project to the more caudal aspects of the sector and rostral areas project to the more rostral areas of the reticular nucleus. There is a large degree of overlap of rostrocaudal projections from the suprasylvian gyrus within the sector, however, the projections originating from rostral sites are situated in a more ventral location compared to the projection originating from the caudal suprasylvian gyrus. Analysis of the distribution of biotin labelled presumptive terminals did not support the notion of 'slabs' or regional variation in terminal density across the mediolateral thickness of the reticular nucleus. In addition, a number of presumptive terminals were found within the internal capsule which coincided with the position of retrogradely labelled cells in the internal capsule following thalamic injections and appears to be part of the perireticular nucleus.The results suggest that the reticular nucleus may be segregated into sectors connected with modality specific cortical areas (e.g. striate and extrastriate visual areas) and nonspecific sectors connected with polymodal (e.g. area 7) cortical regions. The reticular nucleus and its connections with the suprasylvian gyrus may form an important link in binding eye movements to sensory integrative process through visuomotor and auditory thalamic connections.     

The pathways connecting the hippocampal formation, the thalamic reuniens nucleus and the thalamic reticular nucleus in the rat

Most dorsal thalamic nuclei send axons to specific areas of the neocortex and to specific sectors of the thalamic reticular nucleus; the neocortex then sends reciprocal connections back to the same thalamic nucleus, directly as well indirectly through a relay in the thalamic reticular nucleus. This can be regarded as a 'canonical' circuit of the sensory thalamus. For the pathways that link the thalamus and the hippocampal formation, only a few comparable connections have been described. The reuniens nucleus of the thalamus sends some of its major cortical efferents to the hippocampal formation. The present study shows that cells of the hippocampal formation as well as cells in the reuniens nucleus are retrogradely labelled following injections of horseradish peroxidase or fluoro-gold into the rostral part of the thalamic reticular nucleus in the rat. Within the hippocampal formation, labelled neurons were localized in the subiculum, predominantly on the ipsilateral side, with fewer neurons labelled contralaterally. Labelled neurons were seen in the hippocampal formation and nucleus reuniens only after injections made in the rostral thalamic reticular nucleus (1.6-1.8 mm caudal to bregma). In addition, the present study confirmed the presence of afferent connections to the rostral thalamic reticular nucleus from cortical (cingulate, orbital and infralimbic, retrosplenial and frontal), midline thalamic (paraventricular, anteromedial, centromedial and mediodorsal thalamic nuclei) and brainstem structures (substantia nigra pars reticularis, ventral tegmental area, periaqueductal grey, superior vestibular and pontine reticular nuclei). These results demonstrate a potential for the thalamo-hippocampal circuitry to influence the functional roles of the thalamic reticular nucleus, and show that thalamo-hippocampal connections resemble the circuitry that links the sensory thalamus and neocortex.     

A lectin horseradish peroxidase study of the origin of ascending fibers in the medial forebrain bundle of the rat. The upper brainstem

The origins of projections within the medial forebrain bundle from the upper brainstem were examined with the horseradish peroxidase technique. Labeled cells were found in approximately 15 upper brainstem nuclei following injections of a conjugate of horseradish peroxidase and wheat germ agglutinin at various levels of the medial forebrain bundle. Labeled nuclei included (from caudal to rostral): dorsal and ventral parabrachial nuclei; Kolliker-Fuse nucleus; dorsolateral tegmental nucleus; A7 (lateral pontine tegmentum medial to lateral lemniscus); median and dorsal raphe nuclei; distinct group of cells oriented mediolaterally in the dorsal pontine tegmentum below the central gray; B9 (ventral midbrain tegmentum dorsal to medial lemniscus); retrorubral nucleus; nucleus of Darkschewitsch, interfascicular nucleus; rostral and caudal linear nuclei; ventral tegmental area; medial part of substantia nigra, pars compacta; and the supramammillary nucleus. With the exception of the ventral parabrachial nucleus, Kolliker-Fuse, A7, B9 and substantia nigra, pars compacta, each of the nuclei mentioned above sent strong projections along the medial forebrain bundle to the rostral forebrain. Sparse labeling was observed throughout the pontine and midbrain reticular formation. With the exception of the dorsal raphe nucleus, projections to the most anterior regions of the medial forebrain bundle (level of the anterior commissure) essentially only arose from presumed dopamine-containing nuclei-retrorubral nucleus (A8 area), interfascicular nucleus, rostral and caudal linear nuclei, substantia nigra pars compacta, and ventral tegmental area. Evidence was reviewed indicating that major forebrain sites of termination for these dopaminergic nuclei are structures that have been collectively referred to as the 'ventral striatum'. It is concluded from the present findings that several pontine and mesencephalic cell groups are in a position to exert a strong, direct effect on structures in the anterior forebrain and that the medial forebrain bundle is the main communication route between the upper brainstem and the forebrain.     

The afferent connections of the inferior olivary complex in rats: A study using the retrograde transport of horseradish peroxidase

Retrograde transport of horseradish peroxidase (HRP) was used to define the origin of afferents to the inferior olivary complex (IOC) in rats. Using both ventral and dorsal surgical approaches to the brainstem, HRP was injected into the IOC through a micropipette affixed to the tip of a 1-μl Hamilton syringe. After a 2-day postoperative survival, animals were sacrificed by transcardiac perfusion with a 1% paraformaldehyde-1.25% gluteraldehyde solution, and brains were processed according to the DeOlmos protocol (1977), using o-dianisidine as the chromogen. Labeled cells were found at many levels of the nervous system extending from lumbar spinal cord to cerebral cortex. This wide-ranging input from numerous regions clearly underscores the complexity of the IOC and its apparent involvement in several functions. Within the spinal cord, labeled neurons were identified from cervical to lumbar but not at sacral levels. These neurons were found contralaterally in the neck region of the dorsal horn and in the medial portions of the intermediate gray. In the caudal brainstem, reactive cells in the dorsal column nuclei, the spinal trigeminal nucleus, and the subnucleus y of the vestibular complex were observed primarily contralateral to the injection sites. Labeling within the gigantocellular, magnocellular, ventral, and lateral reticular nuclei and the nucleus prepositus hypoglossi was primarily ipsilateral. Reactive neurons in the medial and inferior vestibular nuclei were predominantly ipsilateral or contralateral to HRP injections into the caudal or rostral IOC, respectively. The dentate and interposed nuclei of the cerebellum contained small, lightly labeled neurons primarily contralateral to the injection site, while the fastigial nuclei contained a few relatively large, heavily labeled cells bilateral to caudal olivary injections. Ipsilaterally labeled mesencephalic regions included the periaqueductal gray, interstitial nucleus of Cajal, rostromedial red nucleus, ventral tegmental area, medial terminal nucleus of the accessory optic tract, nucleus of the optic tract, and the lateral deep mesencephalic nucleus. The caudal part of the pretectum and small cells of the stratum profundum of the superior colliculus were labeled predominantly contralateral to the injection. In the caudal diencephalon labeled neurons were most numerous within the nucleus of Darkschewitsch and the subparafascicular nucleus, primarily ipsilateral to olivary injections. Scattered reactive neurons were also found within the ipsilateral zone incerta. With the exception of the zona incerta, all labeled mesencephalic and diencephalic nuclei had some bilateral representation of labeled cells. No labeled neurons were identified within the basal ganglia, while numerous reactive cells were found bilaterally within layer V of the frontal and parietal cerebral cortex.     

Afferents to the rat red nucleus studied by means of D-[3H]aspartate, [3H]choline and non-selective tracers

Following injection of horseradish peroxidase-labeled wheat germ agglutinin or of rhodamine-labeled microspheres as non-selective tracers into the rat red nucleus, the origins of the corticorubral and cerebellorubral pathways, as well as a considerable number of other brain structures including dorsal raphé nucleus, zona incerta and several hypothalamic nuclei showed retrogradely labeled perikarya. Labeling patterns obtained with horseradish peroxidase-labeled wheat germ agglutinin compared well with those observed following application of rhodamine-labeled microspheres which produced injection sites restricted to the small nucleus. In these latter cases, counterstaining with phosphine allowed a better definition of anatomical structures. After D-[3H]aspartate application, retrogradely labeled perikarya were observed in cerebral cortex (layer V), zona incerta, dorsal raphé nucleus and in several other structures also labeled by non-selective tracers. Following application of [3H]choline and using an improved autoradiographic method, perikaryal labeling was massive within nucleus interpositus, while it was absent in dorsal raphé nucleus, cerebral cortex and zona incerta. Retrograde tracing experiments with D-[3H]aspartate and [3H]choline revealed that these transmitter related compounds are selective markers for two subsets of afferents to the red nucleus. The transmitter specificity of the selective labeling with [3H]choline in the cerebellorubral pathway is supported only in part by the results obtained with other methods. The selective labeling with D-[3H]aspartate in the corticorubral pathway, on the other hand, is consistent with its transmitter specificity.