Mamillary body in the rat: topography and synaptology of projections from the subicular complex, prefrontal cortex, and midbrain tegmentum

The retrograde and anterograde transport of horseradish peroxidase conjugated to wheat germ agglutinin (WGA-HRP) has been used to trace afferent connections of the rat mamillary body (MB) at the light and electron microscopic levels. Injections of WGA-HRP into different parts of the MB resulted in heavy retrograde labeling in the subicular complex, medial prefrontal cortex, and dorsal and ventral tegmental nuclei. Injections of WGA-HRP into each of these brain regions, respectively, resulted in anterograde labeling with specific distributions and characteristic synaptic organizations in the MB. Projections from the rostrodorsal and caudoventral subiculum terminated in a topographically organized laminar fashion in the medial mamillary nucleus bilaterally, whereas afferent projections from the presubiculum and parasubiculum terminated only in the lateral mamillary nucleus. Labeled axon terminals which originated from the subicular complex were characterized by round vesicles and formed asymmetric synaptic junctions with small-diameter dendrites and dendritic spines in the medial and lateral mamillary nuclei. Projections from the prefrontal cortex originated mainly in the infralimbic area and to a lesser degree in the prelimbic and anterior cingulate areas. Injections of tracer into these brain regions gave rise to dense labeling of axon terminals in the medial mamillary nucleus, pars medianus, and in the anterior dorsomedial portion of the pars medialis. The labeled terminals were characterized by round vesicles and formed asymmetric synaptic junctions with small-diameter dendrites and dendritic spines. Projections from the dorsal tegmental nucleus terminated in the ipsilateral lateral mamillary nucleus, whereas afferent projections from the anterior and posterior subnuclei of the ventral tegmental nucleus terminated topographically in the medial mamillary nucleus. The ventral tegmental nucleus, pars anterior projected to the midline region of the medial nucleus and the dorsolateral and ventromedial subdivisions of the pars posterior projected to medial and lateral parts of the medial nucleus, respectively. In contrast to the synaptic morphology of subicular complex and medial prefrontal cortex axon terminals in the MB, labeled axon terminals in the MB which originated from the midbrain tegmentum were characterized by pleomorphic vesicles and formed symmetric synaptic junctions with neuronal somata and proximal dendrites as well as distal dendrites and dendritic spines.(ABSTRACT TRUNCATED AT 400 WORDS)     

Convergent prefrontal cortex and mamillary body projections to the medial pontine nuclei: A light and electron microscopic study in the rat

To study the convergence of medial prefrontal cortex and mamillary body projections to the medial pontine nuclei, light and electron microscopic, neuroanatomical, tract-tracing experiments were performed. Injections of horseradish peroxidase conjugated to wheat germ agglutinin (WGA-HRP), biotin conjugated to dextran (BD), or rhodamine conjugated to dextran (RD) were made individually or in combinations into the cerebral cortex, hypothalamus, or pons. In addition, injections of WGA-HRP into the medial prefrontal cortex and electrolytic lesions of the mamillary body were made to study the synaptology of afferent projections to the pontine nuclei. In the light microscopic studies, injections of WGA-HRP into the rostromedial pontine nuclei produced dense, retrograde labeling both in the dorsal peduncular area of the medial prefrontal cortex and in the medial mamillary nucleus, pars medialis. Injections of the anterograde tracers BD and RD into the medial prefrontal cortex and the medial mamillary nuclei, respectively, resulted in partially overlapping terminal fields in the rostromedial pontine nuclei. In the electron microscopic studies, injections of WGA-HRP into the dorsal peduncular area and electrolytic lesions of the mamillary body produced anterogradely labeled axon terminals and degenerating axon terminals that synapsed on the same dendrites or neuronal somata in the rostromedial pontine nuclei. The results demonstrate that the medial prefrontal cortex and the medial mamillary nuclei have partially overlapping projections to the rostromedial pontine nuclei and implicate precerebellar relay nuclei in the integration of limbic and/or autonomic functions mediated by convergent projections from the cerebral cortex and the hypothalamus. J. Comp. Neurol. 398:347–358, 1998. © 1998 Wiley-Liss, Inc.     

Ultrastructural study of the GABAergic and cerebellar input to the nucleus reticularis tegmenti pontis

The nucleus reticularis tegmenti pontis is an intermediate of the cerebrocerebellar pathway and serves as a relay centre for sensorimotor and visual information. The central nuclei of the cerebellum provide a dense projection to the nucleus reticularis tegmenti pontis, but it is not known to what extent this projection is excitatory or inhibitory, and whether the terminals of this projection contact the neurons in the nucleus reticularis tegmenti pontis that give rise to the mossy fibre collaterals innervating the cerebellar nuclei. In the present study the nucleus reticularis tegmenti pontis of the cat was investigated at the ultrastructural level following anterograde and retrograde transport of wheat germ agglutinin coupled to horseradish peroxidase (WGA-HRP) from the cerebellar nuclei combined with postembedding GABA immunocytochemistry. The neuropil of this nucleus was found to contain many WGA-HRP labeled terminals, cell bodies and dendrites, but none of these pre- or postsynaptic structures was double labeled with GABA. The vast majority of the WGA-HRP labeled terminals contained clear spherical vesicles, showed asymmetric synapses, and contacted intermediate or distal dendrites. Many of the postsynaptic elements of the cerebellar afferents in the nucleus reticularis tegmenti pontis were retrogradely labeled with WGA-HRP, while relatively few were GABAergic. We conclude that all cerebellar terminals in the nucleus reticularis tegmenti pontis of the cat are nonGABAergic and excitatory, and that they contact predominantly neurons that project back to the cerebellum. Thus, the reciprocal circuit between the cerebellar nuclei and the nucleus reticularis tegmenti pontis appears to be well designed to function as an excitatory reverberating loop.     

Fine structure of the lateral mammillary projection to the dorsal tegmental nucleus of Gudden in the rat.

The synaptic organization of projections from the lateral mammillary neurons within the dorsal tegmental nucleus of Gudden is studied in the rat with the aid of anterograde transport of horseradish peroxidase conjugated with wheat germ agglutinin (WGA-HRP) and visualized with tetramethylbenzidine. The dorsal tegmental nucleus consists of the pars ventralis (TDV) and the pars dorsalis (TDD). The normal neuropil of the dorsal tegmental nucleus contains three classes of axodendritic terminals, that is, terminals containing round, flat, and pleomorphic vesicles. They make up 44%, 5%, and 51%, respectively, of all axodendritic terminals in the TDV, and 62%, 1%, and 37% in the TDD. Injection of WGA-HRP into the lateral mammillary nucleus permits ultrastructural recognition of many anterograde labeled terminals within both the TDV and TDD. In the TDV, 81% of the labeled terminals contain round synaptic vesicles and make asymmetric synaptic contacts. A few of the labeled terminals contain pleomorphic vesicles and make symmetric synaptic contacts. More than 50% of the labeled terminals contact intermediate dendrites (1-2 microns diameter). In the TDD, almost all labeled terminals are small, contain round vesicles, and make asymmetric synaptic contacts. These terminals mainly contact intermediate as well as distal (less than 1 micron diameter) dendrites. There are only a few labeled terminals with pleomorphic vesicles and no terminals with flat vesicles. The termination pattern of the lateral mammillary neurons in the TDV is similar to that in the TDD. Anterograde labeled axon terminals often contact retrograde labeled dendrites in the TDV. No reciprocal connections are present in the TDD. These results suggest that the TDV and the TDD receive mainly excitatory and a few inhibitory inputs from the lateral mammillary nucleus. The TDV neurons also have direct reciprocal connections with the lateral mammillary neurons.     

Transnuclear transport and axon collateral projections of the mamillary nuclei in the rat

Transnuclear transport of horseradish peroxidase (HRP) and wheat germ agglutinin-HRP conjugate (WGA-HRP) and the retrograde transport of fluorescent tracers were used to study axon collaterals of neurons in the mamillary nuclei. Tracers were injected into the thalamus or brain stem and after 18 hour-5 day survival periods, the brains were processed for fluorescence microscopy or for light and electron microscopic HRP histochemistry. Neurons in all divisions of the ipsilateral mamillary nuclei projected to both the thalamus and tegmentum. After HRP and WGA-HRP injections, anterogradely labeled axon terminals were observed in the known projection fields of the mamillary nuclei. Mamillary neurons were characterized by deeply invaginated, eccentrically located nuclei. Most labeled terminals of axon collaterals in the contralateral anterodorsal thalamic nucleus and dorsal and ventral tegmental nuclei contained round vesicles and formed asymmetrical synapses with somata and dendrites. The present results demonstrate that transnuclear transport of HRP and WGA-HRP can be used to study the connectivity and ultrastructure of axon collaterals and their cells of origin in the central nervous system in a manner comparable to that of transganglionic transport in the peripheral nervous system.     

Ultrastructure of the mammillotegmental projections to the ventral tegmental nucleus of Gudden in the rat.

This study examines the termination pattern of axons from the medial mammillary nucleus within the ventral tegmental nucleus of Gudden (TV) in rats by using anterograde transport of horseradish peroxidase conjugated with wheat germ agglutinin (WGA-HRP) and visualized with tetramethylbenzidine. The neuropil of TV contains three classes of axodendritic terminals, that is, terminals containing round, flat, and pleomorphic synaptic vesicles. These types make up 55.6%, 26.1%, and 18.3%, respectively, of all normal axodendritic terminals. Injection of WGA-HRP into the medial mammillary nucleus permits ultrastructural recognition of anterogradely labeled terminals within the TV. More than 80% of the labeled terminals contain round synaptic vesicles and form asymmetric synaptic contacts, whereas about 16% contain flat synaptic vesicles with symmetric synaptic contacts. There are a few labeled terminals with pleomorphic vesicles and only a few axosomatic terminals. Almost all labeled terminals are small, having diameters of less than 1.5 microns. Compared with the distributions of normal and labeled terminals with round vesicles, there is an increase of the percentage of labeled terminals with round vesicles on the intermediate dendrites (1-2 microns diameter) and a decrease on the distal dendrites (less than 1 micron diameter). Anterogradely labeled axon terminals often contact retrogradely labeled dendrites. These results suggest that the medial mammillary neurons send mainly excitatory as well as a few inhibitory inputs to the dendrites of TV and have direct reciprocal contacts with the TV neurons.     

Retrograde double-labeling study of the mammillothalamic and the mammillotegmental projections in the rat

Collateral axonal branching from the medial or lateral mammillary nuclei to the anterior thalamus, Gudden's tegmental nuclei, the nucleus reticularis tegmenti pontis, and the medial pontine nucleus was studied using the fluorescent retrograde double-labeling method. One day after injection of Fast Blue into the anterior thalamic nuclei or Gudden's tegmental nuclei, Nuclear Yellow was injected into Gudden's tegmental nuclei or the nucleus reticularis tegmenti pontis and the medial pontine nucleus. Following 1 day survival, single- and double-labeled neurons were examined in the mammillary nuclei. The lateral mammillary nucleus contains neurons whose collateral fibers project to both the dorsal tegmental nucleus of Gudden and the ipsilateral or contralateral anterodorsal thalamic nucleus, to both the medial pontine nucleus and the anterodorsal thalamic nucleus, and to both the dorsal tegmental nucleus of Gudden and the medial pontine nucleus. The pars medianus and pars medialis of the medial mammillary nucleus contain neurons whose collateral fibers project to both the anteromedial thalamic nucleus and the ventral tegmental nucleus of Gudden, to both the anteromedial thalamic nucleus and the medial part of the nucleus reticularis tegmenti pontis, and to both the ventral tegmental nucleus of Gudden and the medial part of the nucleus reticularis tegmenti pontis. The dorsal half of the pars posterior of the medial mammillary nucleus contains a few neurons whose collateral fibers project to both the anteromedial thalamic nucleus and the rostral part of the ventral tegmental nucleus of Gudden, and to both the caudal part of the anteroventral thalamic nucleus and the rostral part of the ventral tegmental nucleus of Gudden, while the pars lateralis of the medial mammillary nucleus contains no double-labeled neurons and projects only to the anteroventral thalamic nucleus.     

Visual projections to the pontine nuclei in the rabbit: orthograde and retrograde tracing studies with WGA-HRP

Visual projections to the pontine nuclei in the rabbit were examined by means of both orthograde and retrograde tracing of WGA-HRP. The tecto-pontine projection was examined following microinjections of WGA-HRP in the right superior colliculus. The projection to the pontine nuclei is strictly ipsilateral and terminates at middle and caudal levels of the pons. The projection is absent in rostral pontine nuclei. The strongest projection is to the dorsal border of the dorsolateral pontine nuclei and is the only projection seen when the primary injection site is confined to superficial laminae. When the primary injection site also includes intermediate and deep laminae, patches of labelled terminals are also seen within dorsolateral, lateral, peduncular, paramedian, and ventral pontine nuclei as well as in the contralateral nucleus reticularis tegmenti pontis. The striate corticopontine projection was also examined with orthograde tracing of WGA-HRP. The striate corticopontine projection is ipsilateral. Most labelled terminals were seen in dorsolateral and lateral pontine nuclei throughout the rostral half of pons with some additional terminal labelling in paramedian and peduncular nuclei. Labelled terminals were also seen in ventral pontine nuclei throughout the middle and caudal levels of the pons. In a retrograde tracing study, visual projections to the pontine nuclei were examined following microinjections of WGA-HRP into the pontine nuclei. Labelled cells were seen ipsilaterally in superficial and deep laminae of the superior colliculus and in layer V of striate and surrounding occipital cortex. The pontine nuclei also receive ipsilateral projections from the ventral lateral geniculate, the nucleus of the optic tract, anterior and posterior pretectal nuclei, and the dorsal and medial terminal nuclei of the accessory optic system. These pathways are potential sources of visual input to the cerebellum.     

The GABAergic neurons and axon terminals in the lateralis medialis-suprageniculate nuclear complex of the cat: GABA-immunocytochemical and WGA-HRP studies by light and electron microscopy

In order to get more detailed information on the neural circuit of the lateralis medialis-suprageniculate nuclear (LM-Sg) complex of the cat, the GABAergic innervation of this complex was studied by GABA immunohistochemical techniques. Small immunoreactive cells were found throughout the LM-Sg complex. On the basis of their ultrastructural features, these GABAergic cells were identified as Golgi type II interneurons. The neuropil of this nucleus displayed a conspicuous granular immunoreactivity. Ultrastructurally, the immunoreactive neural profiles in the neuropil were identified as the presynaptic dendrites of interneurons, myelinated axons, or axon terminals. The GABAergic dendritic profiles, containing pleomorphic synaptic vesicles, were involved in synaptic glomeruli. Additionally, GABAergic axon terminals containing pleomorphic synaptic vesicles formed symmetric axodendritic synaptic contacts mainly in the extraglomerular neuropil. They appeared to correspond to either axon terminals from the thalamic reticular nucleus (TRN) or the axon terminals of interneurons. The projections from the TRN to the LM-Sg complex were studied by using wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP). Following injection of WGA-HRP into the LM-Sg complex, a number of retrogradely labeled cells were observed in the TRN. The connections between the TRN and the LM-Sg complex appeared to be topographically organized, the dorsal TRN being connected mainly with the dorsomedial portion of the LM-Sg complex, and the ventral TRN being connected chiefly with the ventrolateral portion of the LM-Sg complex. Following injection of the tracer into the TRN, ultrastructural examination of anterograde labeling in the LM-Sg complex revealed that labeled terminals contain pleomorphic vesicles and make symmetric synaptic contacts mainly with small to medium-sized dendrites. The labeled terminals were not involved in synaptic glomeruli. The present results provide anatomic support for the contention that the projection cells of the LM-Sg complex may be inhibited by both the TRN axons and interneurons, probably through the mediation of GABA.     

Synaptic organization of septal projections in the rat medial habenula: a wheat germ agglutinin-horseradish peroxidase and immunohistochemical study.

The synaptic organization of septal inputs to the rat habenular complex of the dorsal diencephalon was examined employing the anterograde tracer wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). The cellular distribution of substance P (SP) and choline acetyltransferase (ChAT) immunoreactivity was also studied at the light and electron microscopic level. Following placements of tracer within the entire septum, labeled axons were observed in the stria medullaris and in the medial and lateral subnuclei of the habenula. Following injections of tracer in the nuclei triangularis and septofimbrialis of the posterior septum, the medial subnucleus was heavily labeled, whereas the lateral subnucleus was devoid of peroxidase activity. The medial subnucleus possessed labeled myelinated axons and terminals that contained clear, spherical vesicles and formed asymmetric contacts with dendritic spines and shafts. Terminals possessing WGA-HRP activity also formed non-synaptic junctions with other labeled or unlabeled terminals. SP and ChAT immunoreactivity in normal and colchicine-treated animals was confined to dendrites and somata within the medial habenula. Terminals containing clear spherical vesicles formed asymmetric synaptic contacts with these immunoreactive somatic and dendritic profiles. Based on the combined anterograde tracing and immunohistochemical data, it is proposed that septal projections provide a direct innervation to habenular neurons that contain ChAT or SP activity. These septal inputs may play an important role in the facilitation of the ChAT- and SP-positive habenular neurons, both of which provide prominent afferent inputs to the interpeduncular nucleus. Thus, neurons of the habenula and interpeduncular nucleus are under the direct and indirect influence of septal neurons within the limbic forebrain circuit.     

Organization of the projections of a circumventricular organ: the area postrema in the rat

The projections of the rat area postrema were analysed using anterograde and retrograde axonal transport techniques. Discrete injections of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) into the area postrema produced anterograde labeling in specific medullary and pontine nuclei. In the medulla, anterograde labeling was present in the internal solitary zone and dorsal division of the medial solitary nucleus, both of which also contained a small number of retrogradely labeled perikarya. Prominent projections to the dorsal motor nucleus of the vagus were seen only if the WGA-HRP injections in the area postrema invaded dorsal solitary nuclei. In the pons, anterograde labeling was present in the parabrachial nuclei, the dorsolateral tegmental nucleus, and the pericentral division of the dorsal tegmental nucleus. By far the major pontine projection was to the dorsolateral region of the middle one-third of the rostrocaudal extent of the parabrachial nuclei. Retrograde fluorescent tracing studies indicated that most area postrema neurons take part in this parabrachial projection. The area postrema projection to the parabrachial nuclei was bilaterally distributed, whereas that from the dorsal solitary nuclei was primarily ipsilateral. The external solitary zone, immediately subadjacent to the area postrema, neither received area postrema projections nor participated in the projections to the parabrachial nuclei. Fluorescent retrograde double labeling studies confirmed the bilateral nature of the area postrema projection to the parabrachial nuclei. In addition, because no doubly labeled neurons were observed it appears that individual area postrema neurons project to either side but not both sides of the dorsal pons. Thus, numerous neuronal pathways exist for the transfer of blood-borne information (that cannot cross the blood-brain barrier) from the area postrema to other brain regions.     

Mamillary body in the rat: a cytoarchitectonic, Golgi, and ultrastructural study

The present study provides a comprehensive light and electron microscopic analysis of the anatomical organization of the rat mamillary body. The cytoarchitecture and morphology of mamillary neurons were investigated with the aid of Nissl-stained and Golgi-impregnated sections cut in transverse, horizontal, and sagittal planes. The ultrastructural features of the mamillary nuclei were correlated with observations made on Golgi material. The mamillary body is comprised of a lateral and a medial nucleus, the latter being subdivided into five major subnuclei: pars lateralis, pars basalis, pars medialis, pars medianus, and pars posterior. The perikarya are medium-sized or small with the proportions of each differing among subnuclei. The largest perikarya are found in the lateral mamillary nucleus (cell area 257.0 microns2) and have 2-5 radially oriented aspiny dendrites that are often beaded. Small cells predominate in the pars lateralis (cell area 116.3 microns2) and pars basalis (cell area 118.3 microns2), whereas the pars medialis (cell area 196.7 microns2), pars medianus (cell area 136.5 microns2), and pars posterior (cell area 154.6 microns2) contain mainly medium-sized cells. The dendrites of most cells in the medial nucleus are radially oriented and exhibit a variety of spines including numerous short stubby spines, spines with thin necks that end in spherical swellings, and long thin spines. Neuronal somata are often closely apposed with no intervening glial processes and contain eccentrically located nuclei with one or more invaginations of the nuclear envelope. Two main classes of axon terminals were identified in the mamillary body. One type contains round vesicles and forms asymmetric synaptic junctions (RA) with dendrites and dendritic spines. RA terminals rarely contact neuronal somata and proximal dendrites in the MB. The second type contains pleomorphic vesicles and forms mainly symmetric synaptic junctions (PS) with neuronal somata as well as dendrites and spinous processes. Dense-cored vesicles were frequently seen in both types of terminals. Both types of terminals often synapse with two adjacent dendrites and are also found near or adjacent to each other on the same dendrite. A quantitative analysis indicated that the numbers of RA terminals in the medial nucleus almost equals the numbers of PS terminals, whereas the lateral mamillary nucleus contains considerably more PS (64%) than RA terminals (36%).     

Synaptic organization of projections from basal forebrain structures to the mediodorsal thalamic nucleus of the rat.

The synaptic organization of the mediodorsal thalamic nucleus (MD) in the rat was studied with the electron microscope, and correlated with the termination of afferent fibers labeled with wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). Presynaptic axon terminals were classified into four categories in MD on the basis of the size, synaptic vesicle morphology, and synaptic membrane specializations: 1) small axon terminals with round synaptic vesicles (SR), which made asymmetrical synaptic contacts predominantly with small dendritic shafts; 2) large axon terminals with round vesicles (LR), which established asymmetrical synaptic junctions mainly with large dendritic shafts; 3) small to medium axon terminals with pleomorphic vesicles (SMP), which formed symmetrical synaptic contacts with somata and small-diameter dendrites; 4) large axon terminals with pleomorphic vesicles (LP), which made symmetrical synaptic contacts with large dendritic shafts. Synaptic glomeruli were also identified in MD that contained either LR or LP terminals as the central presynaptic components. No presynaptic dendrites were identified. In order to identify terminals arising from different sources, injections of WGA-HRP were made into cortical and subcortical structures known to project to MD, including the prefrontal cortex, piriform cortex, amygdala, ventral pallidum and thalamic reticular nucleus. Axons from the amygdala formed LR terminals, while those from the prefrontal and insular cortex ended exclusively in SR terminals. Fibers labeled from the piriform cortex formed both LR and SR endings. Based on their morphology, all of these are presumed to be excitatory. In contrast, the axons from the ventral pallidum ended as LP terminals, and those from the thalamic reticular nucleus formed SMP terminals. Both are presumed to be inhibitory. At least some terminals from these sources have also been identified as GABAergic, based on double labeling with anterogradely transported WGA-HRP and glutamic acid decarboxylase (GAD) immunocytochemistry.     

Axonal projections and synapses from the supratrigeminal region to hypoglossal motoneurons in the rat

Neural circuits from the supratrigeminal region (Vsup) to the hypoglossal motor nucleus were studied in rats using anterograde and retrograde neuroanatomical tracing methodologies. Iontophoretic injection of 10% biotinylated dextran amine (BDA) unilaterally into the Vsup anterogradely labeled axons and axon terminals bilaterally in the hypoglossal nucleus (XII) as well as other regions of the brainstem. In the ipsilateral XII, the highest density of BDA labeling was found in the dorsal compartment and the ventromedial subcompartment of the ventral compartment, where BDA labeling formed a dense, patchy distribution. Microinjection of 20% horseradish peroxidase (HRP) ipsilaterally or bilaterally into the tongue resulted in retrograde labeling of XII motoneurons confined to the dorsal and ventral compartments of the hypoglossal motor nucleus. Under light microscopical examination, BDA-labeled terminals were observed closely apposing the somata and primary dendrites of HRP-labeled hypoglossal motoneurons. Two hundred and sixty-five of these BDA-labeled terminals were examined at the ultrastructural level. One hundred and twelve BDA-labeled axon terminals were observed synapsing with either the somata (39%, 44/112) or the large or medium-size dendrites (61%, 68/112) of retrogradely labeled hypoglossal motoneurons. Axon terminals containing spherical vesicles (S-type) formed asymmetric synapses with HRP-labeled hypoglossal motoneuron dendrites. In contrast to this, F_F-type axon terminals, containing flattened vesicles, formed symmetric synapses with both the somata and dendrites of HRP-labeled hypoglossal motoneurons with a preponderance of the contacts on their somata. Axon terminals containing pleomorphic vesicles (F_P-type) were noted forming both symmetric and asymmetric synapses with HRP-labeled hypoglossal motoneuron somata and dendrites. The present study provides anatomical evidence of neuronal projections and synaptic connections from the supratrigeminal region to hypoglossal motoneurons. These data suggest that the supratrigeminal region, as one of the premotor neuronal pools of the hypoglossal nucleus, may coordinate and modulate the activity of tongue muscles during oral motor behaviors.     

Gudden's tegmental nuclei and their connections to the hypothalamus and the reticular formation. II. An experimental study using retrograde double labelling with HRP and iron-dextran in the rat

12 rats in double experiments were studied for the simultaneous retrograde transport of HRP and Dextran-fer from injections in the mamillary body and in the reticular formation or in selected diencephalic structures into the cells of Gudden's tegmental nuclei. The results of this particular study corroborated the findings of retrograde transport in single experiments (see Part I) showing all the cells of ventral Gudden's nucleus as projecting into the mamillary body, the cells of its peripheral parts as projecting also via a collateral to the lateral hypothalamus, and the cells of its central parts as projecting, in addition, via a collateral to the reticular formation (ncl. pedunculo-pontinus and ncl. pontis oralis). Cells of the ventral part of the ncl. dorsalis tegmenti Guddeni project into the lateral mamillary body and some of then also through a collateral into the whole of the hypothalamus. Part of its cells project independently into the RF (ncl. pontis oralis). The dorsal part of the ncl. dorsalis tegmenti Guddeni projects bilaterally to the ncl. pontis oralis.     

Input from central nucleus of the amygdala efferents to pericoerulear dendrites,some of which contain tyrosine hydroxylase immunoreactivity

Light microscopic anterograde tracing studies indicate that neurons in the central nucleus of the amygdala (CNA) project to a region of the dorsal pontine tegmentum ventral to the superior cerebellar peduncle which contains noradrenergic dendrites of the nucleus locus coeruleus (LC). However, it has not been established whether the efferent terminals from the CNA target catecholamine-containing dendrites of the LC or dendrites of neurons from neighboring nuclei which may extend into this region. To examine this question, we combined immunoperoxidase labeling of the anterograde tracer biotinylated dextran amine (BDA) from the CNA with immunogold-silver labeling of the catecholamine-synthesizing enzyme tryrosine hydroxylase (TH) in the rostrolateral LC region of adult rats. By light microscopy, BDA-labeled processes were dense in the dorsal pons within the parabrachial nuclei as well as in the pericoerulear region immediately ventral to the superior cerebellar peduncle. Higher magnification revealed that BDA-labeled varicose fibers overlapped TH-labeled processes in this pericoerulear region. By electron microscopy, anterogradely labeled axon terminals contained small, clear as well as some large dense core vesicles and were commonly apposed to astrocytic processes along some portion of their plasmalemma. BDA-labeled terminals mainly formed symmetric type synaptic contacts characteristic of inhibitory transmitters. Of 250 BDA-labeled axon terminals examined where TH immunoreactivity was present in the neuropil, 81% contacted unlabeled and 19% contacted TH-labeled dendrites. Additionally, amygdala efferents were often apposed to unlabeled axon terminals forming asymmetric (excitatory type) synapses. These results demonstrate that amygdaloid efferents may directly alter the activity of catecholaminergic and non-catecholaminergic neurons in this pericoerulear region of the rat brain. Furthermore, our study suggests that CNA efferents may indirectly affect the activity of pericoerulear neurons through modulation of excitatory afferents. Amygdaloid projections to noradrenergic neurons may help integrate behavioral and visceral responses to threatening stimuli by influencing the widespread noradrenergic projections from the LC. © 1996 Wiley-Liss, Inc.     

Afferent projections to the interpeduncular nucleus in the rat, as studied by retrograde and anterograde transport of wheat germ agglutinin conjugated to horseradish peroxidase

We examined the afferent projections to the subnuclei of the interpeduncular nucleus (IPN) in the rat by means of retrograde and anterograde transport of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). We observed locations of retrogradely labeled cells following injections of WGA-HRP into the IPN, and distributions of anterogradely labeled fibers and terminals within the IPN following injections into the areas that contain cells of origin of afferents. Results of the retrograde and anterograde experiments have clarified the detailed organization of the IPN afferents. A part of the nucleus incertus, located dorsomedial to the dorsal tegmental nucleus, projects to the contralateral half of the rostral subnucleus of the IPN; the pars caudalis of the dorsal tegmental nucleus projects sparsely to the rostral lateral, dorsal lateral, lateral, caudal, and apical subnuclei predominantly contralaterally; the laterodorsal tegmental nucleus, to most of the subnuclei predominantly contralaterally; the ventromedial central gray rostral to the dorsal tegmental nucleus and lateral to the dorsal raphe nucleus projects to the rostral lateral and dorsal lateral subnuclei predominantly contralaterally; the median raphe nucleus, substantially to all subnuclei; the medial habenular nucleus, in a topographic manner, to the rostral, central, and intermediate subnuclei, to the rostral lateral and lateral subnuclei predominantly ipsilaterally, and to the dorsal lateral subnucleus predominantly contralaterally; the supramammillary nucleus and areas around the origin of the mammillothalamic tract and near the third ventricle project sparsely to the ventral part of the rostral subnucleus and to the central, lateral, caudal and apical subnuclei; the nucleus of the diagonal band, sparsely to the rostral, central, dorsal lateral, caudal, and apical subnuclei. These differential projections of the afferents to the subnuclei of the IPN may reflect its complex functions within the limbic midbrain circuit.     

Ultrastructure and synaptic organization of axon terminals from brainstem structures to the mediodorsal thalamic nucleus of the rat.

The ultrastructural characteristics and synaptic organization of afferent terminals from the brainstem to the mediodorsal thalamic nucleus (MD) of the rat have been studied with the electron microscope, by means of anterograde transport of wheat germ agglutinin-horseradish peroxidase (WGA-HRP). Labeled fibers were seen predominantly in the lateral portion of MD after the injections of WGA-HRP into the substantia nigra pars reticulata (SNr), the superior colliculus (SC), and the dorsal tegmental region (DT). The boutons arising from the SC were relatively small (less than 1.5 microns in diameter), formed asymmetric synaptic contacts with small dendrites and dendritic spines, and contained round synaptic vesicles. The axon terminals from the DT were mostly large boutons (2-4.5 microns) with asymmetric synaptic specializations and round vesicles. These boutons and their postsynaptic targets formed synaptic glomeruli that were entirely or partially ensheathed by glial lamellae. The ultrastructural features are almost identical to those of boutons in the medial and central segments of MD that were previously shown to originate from the basal amygdaloid nucleus and the piriform cortex. The boutons from the SNr had a wide range in size, but the majority were medium-sized to large (1.5-4 microns). The nigral boutons established symmetric synaptic contacts with dendritic shafts and occasionally with somata, and contained pleomorphic vesicles. However, like the DT terminals, they participated in glomerular formations. The nigral terminals closely resemble previously described terminals in the medial part of MD from the ventral pallidum, except that the nigral terminals formed en passant and axosomatic synapses as well as axodendritic synapses. A combined immunohistochemistry and WGA-HRP tracing study revealed that the nigral inputs were immunoreactive for glutamic acid decarboxylase and the axon terminals from the DT were immunoreactive for choline acetyltransferase. In a separate study, the colliculothalamic fibers have been shown to take up and transport the transmitter specific tracer [3H]-D-aspartate, and are therefore putatively glutamatergic and/or aspartatergic. Taken together with this, the present results suggest that the collicular afferents are excitatory and glutamatergic and/or aspartatergic, that the inputs from the DT are also excitatory and cholinergic, while the nigral inputs are inhibitory and GABAergic.     

Origin of brain stem and temporal cortical afferent fibers to the septal region in the squirrel monkey

The origins of the brain stem and temporal cortical projections to the septal region in the squirrel monkey were investigated with the horseradish peroxidase (HRP) retrograde axonal transport technique. After HRP injections placed into the septal region, labeled cells were observed in brain stem sites which generally correspond to regions which are associated with known monoamine cell groups previously identified in the primate. These structures include the nucleus locus ceruleus, dorsal tegmental nucleus of Gudden, nucleus reticularis tegmenti pontis, nucleus annularis, ventral tegmental region, and the medial aspect of the lateral hypothalamus. Temporal cortical efferent fibers to the septal region arise principally from layers II and III of the perirhinal region, suggesting the presence of a second-order olfactory innervation of this structure.     

Prefrontal cortical efferents in the rat synapse on unlabeled neuronal targets of catecholamine terminals in the nucleus accumbens septi and on dopamine neurons in the ventral tegmental area.

Physiological and pharmacological studies indicate that descending projections from the prefrontal cortex modulate dopaminergic transmission in the nucleus accumbens septi and ventral tegmental area. We investigated the ultrastructural bases for these interactions in rat by examining the synaptic associations between prefrontal cortical terminals labeled with anterograde markers (lesion-induced degeneration or transport of Phaseolus vulgaris leucoagglutinin; PHA-L) and neuronal processes containing immunoreactivity for the catecholamine synthesizing enzyme, tyrosine hydroxylase. Prefrontal cortical terminals in the nucleus accumbens and ventral tegmental area contained clear, round vesicles and formed primarily asymmetric synapses on spines or small dendrites. In the ventral tegmental area, these terminals also formed asymmetric synapses on large dendrites and a few symmetric axodendritic synapses. In the nucleus accumbens septi, degenerating prefrontal cortical terminals synapsed on spiny dendrites which received convergent input from terminals containing peroxidase immunoreactivity for tyrosine hydroxylase, or from unlabeled terminals. In single sections, some tyrosine hydroxylase-labeled terminals formed thin and punctate symmetric synapses with dendritic shafts, or the heads and necks of spines. Close appositions, but not axo-axonic synapses, were frequently observed between degenerating prefrontal cortical afferents and tyrosine hydroxylase-labeled or unlabeled terminals. In the ventral tegmental area, prefrontal cortical terminals labeled with immunoperoxidase for PHA-L were in synaptic contact with dendrites containing immunogold reaction product for tyrosine hydroxylase, or with unlabeled dendrites. These results suggest that: (1) catecholaminergic (mainly dopaminergic) and prefrontal cortical terminals in the nucleus accumbens septi dually synapse on common spiny neurons; and (2) dopaminergic neurons in the ventral tegmental area receive monosynaptic input from prefrontal cortical afferents. This study provides the first ultrastructural basis for multiple sites of cellular interaction between prefrontal cortical efferents and mesolimbic dopaminergic neurons.