Estrogen receptor-alpha immunoreactive neurons in the brainstem and spinal cord of the female rhesus monkey: species-specific characteristics.

The distribution pattern of estrogen receptors in the rodent CNS has been reported extensively, but mapping of estrogen receptors in primates is incomplete. In this study we describe the distribution of estrogen receptor alpha immunoreactive (ER-alpha IR) neurons in the brainstem and spinal cord of the rhesus monkey. In the midbrain, ER-alpha IR neurons were located in the periaqueductal gray, especially the caudal ventrolateral part, the adjacent tegmentum, peripeduncular nucleus, and pretectal nucleus. A few ER-alpha IR neurons were found in the lateral parabrachial nucleus, lateral pontine tegmentum, and pontine gray medial to the locus coeruleus. At caudal medullary levels, ER-alpha IR neurons were present in the commissural nucleus of the solitary complex and the caudal spinal trigeminal nucleus. The remaining regions of the brainstem were devoid of ER-alpha IR neurons. Spinal ER-alpha IR neurons were found in laminae I-V, and area X, and were most numerous in lower lumbar and sacral segments. The lateral collateral pathway and dorsal commissural nuclei of the sacral cord and the thoracic intermediolateral cell column also contained ER-alpha IR neurons. Estrogen treatment did not result in any differences in the distribution pattern of ER-alpha IR neurons. The results indicate that ER-alpha IR neurons in the primate brainstem and spinal cord are concentrated mainly in regions involved in sensory and autonomic processing. Compared with rodent species, the regional distribution of ER-alpha IR neurons is less widespread, and ER-alpha IR neurons in regions such as the spinal dorsal horn and caudal spinal trigeminal nucleus appear to be less abundant. These distinctions suggest a modest role of ER-alpha in estrogen-mediated actions on primate brainstem and spinal systems. These differences may contribute to variations in behavioral effects of estrogen between primate and rodent species.

The fine structure of laminae IV, V, and VI of the Macaque spinal cord.

The normal fine structure of the deep dorsal horn (laminae IV, V, and VI) of the lumbosacral spinal cords of adult Macaque monkeys was examined. Axonal profiles with rounded synaptic vesicles (R) constitute about one-fourth of the total synaptic population in lamina IV and gradually increase in number to comprise more than a third of the synaptic population in lamina VI. Conversely, profiles with flattened vesicles (F) make up two-thirds of the synaptic population in lamina IV and slightly more than half in lamina VI. Axons which form the central profile (C) of synaptic glomeruli are more common in laminae IV and V than in lamina VI, and constitute about 5% of the total synaptic population. Profiles with large granular vesicles (LGV) are relatively uncommon in all the deep laminae. The vast majority of synaptic contacts are axodendritic. Axoaxonal synapses are formed between F, and R or C profiles, the F profile being the presynaptic component. It is concluded that there are both light and electron microscopic morphological differences among the deep laminae, as well as between the deep laminae and the superficial laminae of the spinal cord. Therefore, it appears that Rexed's original schema for laminar organization of the dorsal horn in the cat is also applicable to that of the monkey. In view of the relative paucity of LGV profiles in the deep dorsal horn, it is suggested that synapses in these laminae which contain monoamines or peptides are more likely to be associated with clear synaptic vesicles than granular synaptic vesicles.

Serotonin-containing structures in the nucleus raphe dorsalis of the cat: an ultrastructural analysis of dendrites, presynaptic dendrites, and axon terminals.

In the nucleus raphe dorsalis of the cat, an electron microscopic immunocytochemistry method was used to identify the fine structure of serotoninergic dendritic profiles and axon terminals analyzed in serial sections. Two classes of serotoninergic dendrites were distinguished in the nucleus. The first class was constituted by conventional serotonin (5-HT) dendrites that were contacted by unlabeled axon terminals containing differing populations of synaptic vesicles. The second class consisted of serotoninergic dendrites that contained vesicles in their dendritic shafts. Such 5-HT dendrites were further subdivided into two groups according to their synaptic contacts. In some 5-HT vesicle-containing dendrites, the vesicles were densely packed in small clusters and were associated with a well-defined synaptic specialization. These dendrites were classified as serotoninergic presynaptic dendrites and established synaptic contacts with unlabeled and labeled dendrites and were contacted by unlabeled axon terminals. In other 5-HT vesicle-containing dendrites, extensive serial section examination showed that the vesicles could be observed near the membrane but were never found to be associated with any synaptic membrane specialization. Serotoninergic axon terminals that were presumed to be recurrent collaterals of 5-HT neurons were present in the nucleus. Some of them were observed in synaptic contact with dendrites or dendritic protrusions whereas others did not exhibit synaptic specializations. The existence of serotoninergic dendrodendritic synaptic contacts and axon terminals suggests direct local interactions between serotoninergic neurons within the nucleus raphe dorsalis.

Fine structure of calcitonin gene-related peptide immunoreactive synaptic contacts in the thalamus of the rat.

Recent studies have shown a prominent calcitonin gene-related peptide immunoreactive (CGRP-ir) pathway extending from the external medial and external lateral para-brachial nuclei to the area surrounding and including the gustatory nuclei in the thalamus, and the cortex and amygdala. The function of the CGRP-ir pathway is not completely understood, but may be involved with the processing of both nociceptive and gustatory information in the thalamus. The purpose of this study was to characterize the nature of the CGRP-ir synaptic contacts in the gustatory nucleus. Electron microscopic examination of CGRP-ir synaptic contacts revealed two classes of CGRP-ir terminals. One class, which was large, formed asymmetric synaptic contacts on dendritic appendages, had many small, round synaptic vesicles, and heavy patches of reaction product which obscured any underlying organelles. Since similar terminals in unstained tissue contained large numbers of dense-cored vesicles, it was concluded that CGRP-ir was contained predominantly in dense-cored vesicles. A second class of CGRP-ir terminals was smaller and made either asymmetric or symmetric synaptic contacts. Both symmetric and asymmetric small terminals contained small, round synaptic vesicles and fewer patches of dense reaction product. Several of the CGRP-ir terminals making symmetric contacts also contained pleomorphic vesicles. There were very few contacts on cell bodies. There were no contacts on other CGRP-ir elements, somal or dendritic, or on axon terminals. None of the CGRP-ir terminal elements were postsynaptic to unlabeled terminals. Axons containing CGRP-ir were primarily unmyelinated, but a few myelinated axons were also seen.

The distribution of dorsal root axons to laminae IV, V, and VI of the Macaque spinal cord: a quantitative electron microscopic study.

The projections of dorsal root axons to the deeper laminae (IV, V, and VI) of the Macaque spinal cord were examined by the use of experimentally induced degeneration following dorsal rhizotomy or by injection of dorsal root ganglia with tritiated amino acids followed by light and electron microscopic autoradiography. Following dorsal rhizotomy, neurofilamentous degeneration of synaptic profiles occurs in each of the three deep laminae, more commonly in laminae IV and V than in lamina VI. The neurofilamentous degeneration is seen both in central glomerular (C) profiles and in many of the round vesicle (R) profiles. Neurofilamentous degeneration occurs as early as 18 hours following rhizotomy and the degenerating terminals are most numerous at 3-4 days postrhizotomy. None are seen after 7 days survival. The neurofilamentous profiles form axodendritic and, occasionally, axosomatic synapses with neurons of the dorsal horn. They are also seen to be postsynaptic to flat vesicle (F) profiles in axoaxonal synapses. A second type of degeneration, electron-lucent degeneration, is seen in laminae V and VI, and only occasionally in lamina IV. The lucent degeneration occurs somewhat later after rhizotomy than does the neurofilamentous degeneration and reaches its peak at 5 days postrhizotomy. No lucent terminals are seen after 7 days survival. Electron-dense degeneration, so common in lamina II, is not seen in the deeper dorsal horn. Autoradiographic techniques show that both C and R terminals are labelled in the deeper dorsal horn. Both of these terminals form axodendritic synapses and a significant number are found to be postsynaptic in axoaxonal synapses. Most of the C terminals degenerate following rhizotomy or are labelled following injection of the parent dorsal root ganglia with tritiated amino acids. Approximately one-fifth of the R profiles are derived from dorsal roots. F profiles do not appear to be of dorsal root origin in any case. It is concluded that neurofilamentous alterations represent the degeneration of larger-diameter (A beta) axons which distribute to the deeper dorsal horn and that electron-lucent degeneration represents the termination of A delta fibers. Electron-dense degeneration thought to represent the termination of nonmyelinated axons (C fibers) in the superficial dorsal horn is not seen in the deeper dorsal and it is concluded that C fibers do not project to the deeper laminae.

The terminations of corticospinal tract axons in the macaque monkey.

This study examined the corticospinal tract in monkey by utilizing the anterograde transport of wheat germ lectin conjugated to horseradish peroxidase (WGA HRP) at the light microscopic level and the axonal transport of 3H-proteins with both light and electron microscopic autoradiographic techniques. The animals survived 3-9 days after the injections of 3H-leucine or 3H-leucine/WGA HRP into either motor or sensory cortices. With the laminar schema of Rexed as a guide to the layers of the spinal gray matter, qualitative and quantitative analyses of labeled projections of the corticospinal tract (CST) were made. With the light microscope, axons from the sensory cortex labeled with WGA-HRP could be observed in the contralateral spinal gray from lamina I to the border of laminae VI/VII, the heaviest distribution being located in medial III-VI. There was a small ipsilateral projection to V and VI. With 3H label, laminae I and II revealed few overlying silver grains; many grains overlay laminae III-VI. Projections from the motor cortex labeled with either WGA-HRP or 3H extended from the contralateral laminae III/IV border into the motor nucleus (lamina IX) and were seen to be somewhat more dense in the lateral areas of the spinal gray. The motor cortex projected heavily to ipsilateral VIII, and in sparse amounts to ipsilateral V and VI. Electron microscopy of radioactive axons from the sensory cortex to dorsal horn revealed many radioactive myelinated fibers and some labeled non-myelinated axons. Labeled terminals contacted medium to small dendrites; there were a few labeled C-type profiles in glomeruli and occasional axo-axonal or dendro-axonal contacts, the labeled cortical axons being the postsynaptic structure. In ventral horn following motor cortex injections, the labeled axons were all myelinated. The synaptic contacts were found on small, medium, and large proximal dendrites as well as on cell bodies. Labeled terminals which formed the central element in glomeruli were also seen in this region. Most of the labeled corticospinal terminals in dorsal and ventral horn contained rounded vesicles, but a significant number revealed pleomorphic vesicles. The relationship of these morphological findings to physiological studies of the CST is presented.

Light and electron microscopic evidence of transneuronal labeling with WGA-HRP to trace somatosensory pathways to the thalamus.

Horseradish peroxidase conjugated to wheat-germ lectin is being used with increasing frequency as an anterograde label to trace pathways in the nervous system, owing to the sensitivity of the method and ease of use. However, it has been suggested that horseradish peroxidase conjugated to wheat-germ lectin may be transneuronally transported, thus affecting the ease of interpretation of the results. The present study used the projections of the dorsal column nuclei and spinal cord to the thalamus as a model system to determine whether transneuronal transport could be demonstrated and whether the degree of such transport was related to the size of the injection site. Light microscopic observation of sections incubated with tetramethyl benzidine after large injections (1 microL of a 10% solution of horseradish-peroxidase-conjugated wheat-germ lectin in water) in the dorsal column nuclei demonstrated the presence of labeled neurons in the nucleus reticularis thalami, which is not known to receive afferents from or project to these nuclei. The electron microscopic study, although based upon the use of the chromogen benzidine dihydrochloride, less sensitive than tetramethyl benzidine, revealed the existence of labeled neurons in the thalamic ventrobasal complex. This is unlikely to be due to retrograde labeling and is therefore interpreted as a result of transneuronal, perhaps transsynaptic, transport. Glial and perivascular cells also contained granules of reaction product in some cases. Smaller injections (100 nL) in the dorsal column nuclei, on the other hand, did not produce this apparent transneuronal labeling. After small injections (100 nL) in the spinal cord, anterograde labeling was observed mainly in the thalamic ventrobasal complex in the rat, and in the posterior group in the cat, and the nuclei centralis lateralis and submedius in both species, as has been described in numerous other studies. After large injections, additional labeled areas were observed in the posterior intralaminar region (parafascicular-center median complex), in the medial thalamus (nuclei reuniens, rhomboid and paraventricular), and in the cat, in the ventroposterolateral nucleus. In the rat, experiments were performed in which a kainic acid injection was made to induce neuronal loss in the nucleus reticularis gigantocellularis of the medulla, which is a relay of the spinoreticulothalamic pathway, known to project to some of these thalamic areas.(ABSTRACT TRUNCATED AT 400 WORDS)

Ultrastructural analysis of GABA-immunoreactive elements in the monkey thalamic ventrobasal complex.

This study describes the ventrobasal complex of the primate by using GABA immunocytochemistry at the electron microscopic level. The primate ventrobasal complex has a similar synaptic organization to sensory thalamic nuclei in other species. Two synaptic profiles within the ventrobasal complex contain flattened or pleomorphic synaptic vesicles and are GABA-immunoreactive. F-boutons (= F1 type, Guillery's classification; Guillery: Z. Zellforsch. 96:1-38, '69) are located principally in the extraglomerular neuropil and contain densely packed flattened synaptic vesicles and several elongate mitochondria and establish symmetric (Gray's type II) synaptic contacts. These boutons are not found postsynaptic to any other element and are presynaptic principally to nonimmunoreactive elements that are thought to be thalamocortical relay cell dendrites. PSD-boutons (= F2 type, Guillery's classification) contain a moderate number of flattened or pleomorphic synaptic vesicles and fewer mitochondria than F-boutons. PSD-boutons are found in glomerular and extraglomerular areas of neuropil and establish symmetric synaptic contacts. These boutons are considered to be appendages of interneuron dendrites and are postsynaptic to RL-, RS (Guillery's classification)-, F-, and other PSD-boutons. PSD-boutons are presynaptic to thalamocortical relay neurons and interneuron dendrites including PSD-boutons. Problems in distinguishing F- from PSD-boutons are addressed by comparing immunostained and nonimmunostained material and by the use of serial sections. The majority of synaptic contacts between pleomorphic vesicle-containing profiles appear to be between PSD-boutons and other components of interneurons. Few contacts between F-boutons and local circuit neurons are seen. These data suggest the principal GABAergic input to interneurons in the primate ventrobasal complex is derived from other interneurons.

Monosynaptic projections from the nucleus retroambiguus to motoneurons supplying the abdominal wall, axial, hindlimb, and pelvic floor muscles in the female rhesus monkey.

The nucleus retroambiguus (NRA) consists of premotor neurons in the caudal medulla. It is involved in expiration, vomiting, vocalization, and probably reproductive behavior by means of projections to distinct motoneuronal cell groups. Because no information is available about the NRA and its efferent pathways in primates, the present study examines NRA projections to the lumbosacral spinal cord in female rhesus monkeys. To identify the NRA, wheat germ agglutinin-horseradish peroxidase (WGA-HRP) was injected into the lumbosacral cord in three monkeys. To study the distribution of NRA axons in the lumbosacral cord, WGA-HRP injections were made into the NRA in seven monkeys. To identify motoneuronal cell groups receiving input from the NRA, the same seven monkeys also received cholera toxin subunit b (CTb) injections into different hindlimb, axial, and pelvic floor muscles. The results show that NRA neurons projecting to the lumbosacral cord are mainly located between 1 to 4 mm caudal to the obex. They send numerous axons to external oblique and pelvic floor motoneurons, whereas projections to iliopsoas and axial motoneurons are less numerous. The projections are bilateral, but show a clear contralateral predominance in the iliopsoas, axial, and pelvic floor motoneuronal cell groups. At the ultrastructural level, NRA-terminal profiles make asymmetrical contacts with labeled and unlabeled dendrites in these motoneuronal cell groups and contain large amounts of spherical and a few dense core vesicles. It is concluded that the NRA is well developed in the monkey and that there exists a direct pathway from the NRA to lumbosacral motoneurons in this species. The finding that the NRA projects to a somewhat different set of motoneuronal cell groups compared with other species fits the concept that it is not only involved in expiration-related activities but also in species specific receptive and submissive behavior.

Monosynaptic projections from the lateral periaqueductal gray to the nucleus retroambiguus in the rhesus monkey: implications for vocalization and reproductive behavior.

The periaqueductal gray (PAG) is known to be essential for vocalization and reproductive behavior. The PAG controls components of these behaviors by means of projections to the nucleus retroambiguus (NRA), a group of premotor neurons in the caudal medulla oblongata. In the accompanying study (VanderHorst et al., 2000 [accompanying study]), the NRA and its lumbosacral projections have been identified in the rhesus monkey. The present light and electron microscopical tracing study describes the PAG-NRA pathway in primates. To locate midbrain neurons projecting to the NRA, wheat germ agglutinin horseradish peroxidase (WGA-HRP) was injected into the NRA in six monkeys. To determine the distribution pattern of PAG axons in the medulla oblongata, WGA-HRP was injected into the PAG and adjacent tegmentum in three additional monkeys. In one of these three monkeys, biotinylated dextran amine and cholera toxin subunit b were injected into the lumbosacral cord to retrogradely identify NRA neurons. The results show that a compact group of neurons in the medial part of the lateral PAG at the intercollicular level sends a dense projection to the NRA. The projection is bilateral with a clear ipsilateral predominance. At the ultrastructural level, there are monosynaptic contacts between PAG fibers and NRA neurons, including NRA neurons that project to the lumbosacral cord. The synaptic contacts were primarily asymmetrical and the labeled terminal profiles contained spherical and dense core vesicles. It is concluded that there exists a strong and direct PAG-NRA pathway in the rhesus monkey. Because NRA neurons projecting to the lower lumbar cord are included, the PAG-NRA projection is likely to be involved not only in vocalization but also in other behaviors, such as receptive posture.

Bulbospinal projections in the primate: a light and electron microscopic study of a pain modulating system.

The projections of the nucleus raphe magnus (NRM) and the immediately adjacent reticular formation were studied in the macaque monkey following injections of the rostroventral medulla with 3H-leucine and examination of the resultant labeled axons and terminals by light and electron microscopic autoradiography. Five monkeys had accurately placed injections, which resulted in fiber pathway labeling that coursed caudally, laterally, and dorsally to project to laminae I, II, and V of subnucleus caudalis of the trigeminal and then traveled in the dorsolateral funiculus of the cord and terminated in similar laminae of the spinal dorsal horn at cervical levels. The pathway was only lightly labeled caudal to the cervical enlargement and could not be readily discerned above background in the thoracic or lumbar cord. Electron microscopy revealed that axons and terminals serving this system constitute a heterogeneous population. Large-diameter myelinated axons (3-6-micron diameter), small myelinated axons (0.75-3-micron diameter), and clusters of nonmyelinated axons were labeled. Terminals in laminae I, II, and V contained mixtures of clear round and granular vesicles or clear pleomorphic and granular vesicles or formed the central element in synaptic glomeruli. The labeled profiles formed asymmetrical or symmetrical synapses on medium and small dendrites; labeled axosomatic synapses were not observed. In rare instances there were contacts between labeled profiles and vesicle-containing structures, which were probably dendritic, but whether the NRM axon was pre- or postsynaptic to such structures could not be determined. It was concluded that the NRM in the monkey is organized in a manner quite similar to that previously described in the cat. The wide variety of fiber types and synaptic terminals serving this system suggests that different classes of neurons participate in it, probably using several transmitter substances that result in varying postsynaptic effects on neurons located in the trigeminal complex and dorsal horn.

Architecture of individual dendrites from intracellularly labeled thalamocortical projection neurons in the ventral posterolateral and ventral posteromedial nuclei of cat.

This study provides quantitative descriptions of individual dendrites from electrophysiologically characterized and intracellularly labeled thalamocortical projection (TCP) neurons of the cat ventrobasal complex. One hundred nine dendrites from six ventral posterolateral (VPL) neurons and six ventral posteromedial (VPM) neurons were examined. Measurement of several parameters showed that the individual dendrites were very similar to each other in overall architecture even though they varied greatly in total length and number of dendritic branches. The mean path distance (length from soma to a dendritic tip) was very similar for all dendrites in each group (VPL or VPM) regardless of the number of branches found along the path distance. However, VPL dendrites had a longer mean path distance (VPL = 206 +/- 36 microns; n = 51) than VPM dendrites (VPM = 182 +/- 29 microns; n = 58; P < 0.001). For all dendrites there was a strong correlation between the stem dendrite diameter and the dendritic length, which allows the estimation of dendritic length from dendrite diameter. Analysis of dendritic scaling shows that branches higher than first order do not follow Rall's 3/2 power rule, so these neurons cannot be modeled using the equivalent cylinder approximation. The data add to the qualitative descriptions of cat ventrobasal (VB) TCP dendrites currently available and provide a basis for future comparative, developmental, and plasticity studies. Analysis shows that many parameters of cat VB TCP dendrites fall within a narrow range, suggesting that, regardless of differences in length or superficial appearance, these dendrites share a stable underlying architecture.

Transneuronal changes of the inhibitory circuitry in the macaque somatosensory thalamus following lesions of the dorsal column nuclei.

The inhibitory circuitry of the ventroposterolateral nucleus (VPL) of the macaque somatosensory thalamus was analyzed in normal animals and in those surviving for a few days or several weeks following a unilateral lesion of the cuneate nucleus, the source of medial lemniscal (ML) axons carrying information from the contralateral upper extremity. Inhibitory synaptic terminals in the VPL were defined as those that contain flattened or pleomorphic synaptic vesicles and that can be shown to be immunoreactive for gamma-aminobutyric acid (GABA). There are two types of these profiles: F axon terminals that arise from neurons of the thalamic reticular nucleus, and perhaps from VPL local circuit neurons (LCNs); and the dendritic appendages of LCNs that form presynaptic dendrites (PSDs). ML terminals normally have extensive synaptic interactions with PSDs but not with F axon terminals. Electron microscopic analyses revealed that cuneatus lesions resulted in a rapid loss of ML terminals and a statistically significant reduction in both F and PSD synaptic profiles. Confocal scanning microscopy also demonstrated a profound loss of GABA immunoreactivity in the deafferented VPL. These changes persisted for more than 20 weeks, without any evidence of reactive synaptogenesis of surviving sensory afferents or of inhibitory synapses. The changes in GABA circuitry are transneuronal, and the possible mechanisms that may underlie them are discussed. It is suggested that the altered GABAergic circuitry of the VPL in the monkey may serve as a model for understanding changes in somatic sensation in the human following peripheral or central deafferentation.

Analysis of gamma-aminobutyric acidergic synaptic contacts in the thalamic reticular nucleus of the monkey.

Gamma-aminobutyric acidergic (GABAergic) neurons in the thalamic reticular nucleus (TRN) spontaneously generate a synchronous bursting rhythm during slow-wave sleep in most mammals. A previous study at the electron microscopic level in cat anterior TRN has suggested that synchronous bursting activity could result from the large number of presumably GABAergic dendrodendritic synaptic contacts. However, little is known about the synaptology of the monkey thalamic reticular nucleus and whether it contains dendrodendritic contacts. To address this issue, we examined tissue obtained from Macaca fascicularis that was prepared for electron microscopy using postembedding techniques to demonstrate GABA immunoreactivity. Examination of the anterior (motor) and posterior (somatosensory) portions of the TRN disclosed the following: The majority of synaptic contacts (87.5% of 958) were formed by axon terminals showing no GABA immunoreactivity and making asymmetric synaptic contacts on dendrites or cell bodies. A further 6.4% of synaptic contacts was composed of GABA-immunoreactive presynaptic terminals making symmetric contacts with the dendrites of TRN neurons. The majority resembled the pleomorphic vesicle containing F-terminals seen in the dorsal thalamus and known to originate from axons of TRN. A subset or possible second class did not resemble any previously described class of GABA-immunoreactive terminals in the TRN. Both classes of these terminals making symmetric contacts may originate wholly or partially within the nucleus. There was one dendrodendritic synaptic contact and only a small number (3.2%) of axodendritic contacts with synaptic vesicles visible both pre- and postsynaptically. We conclude that dendrodendritic contacts are probably not responsible for the synchronized bursting neuronal activity seen in the slow-wave sleep of monkeys, and that, if TRN neurons are coupled synaptically, the most likely mechanism is through the synapses formed by recurrent axon collaterals of TRN neurons onto TRN dendrites.

Chronic dyspnea and hyperventilation in an awake patient with small subcortical infarcts.

A 79-year-old woman presented with chronic dyspnea and hyperventilation. There was no evidence of pulmonary disease. Hyperventilation persisted during sleep and after high-dose administration of a narcotic. A head MRI revealed bilateral medial thalamic infarctions. Central neurogenic hyperventilation was diagnosed in this alert patient. The case may illustrate a role for the thalamus in regulating ventilation, but another small infarct not visible on MRI also could be responsible.

The differential synaptic organization of the spinal and lemniscal projections to the ventrobasal complex of the rat thalamus. Evidence for convergence of the two systems upon single thalamic neurons.

The synaptic organization of terminals originating either from the spinal cord (spinothalamic) or from the dorsal column nuclei (lemniscal) was investigated in the ventrobasal complex of the rat thalamus. Wheatgerm agglutinin conjugated to horseradish peroxidase was used as an anterogradely transported axonal tracer, using benzidine dihydrochloride as a chromogen for the identification by electron microscopy of spinal and lemniscal projections to the ventrobasal thalamus. A double anterograde tract tracing strategy, based labeling by wheatgerm agglutinin conjugated to horseradish peroxidase of spinal terminals and simultaneous visualization of lemniscal terminals identified by Wallerian degeneration induced by lesion of the neurons of origin in the dorsal column nuclei, was used to compare the postsynaptic elements contacted by the two pathways and to look for a possible convergence of the two pathways onto single thalamic neurons. Spinal and lemniscal terminals are large (2-2.5 microns mean average diameter) terminals containing several mitochondria and numerous rounded vesicles. A quantitative analysis of the mean average diameters of the terminals revealed that one could not differentiate between synapses formed by the two pathways on a morphological basis. Terminals of the two pathways make asymmetrical contacts (Gray type I) with dendrites of varying diameter, dendritic protrusions, and cell somata. A quantitative analysis of the least diameter of the postsynaptic elements demonstrates projections of the two systems to different, partially overlapping regions of thalamic neurons. Lemniscal terminals originating from the dorsal column nuclei frequently contact cell somata; axosomatic spinothalamic contacts are uncommon. In addition, lemniscal projections tend to contact more proximal dendrites than do spinal projections, and this differential synaptic organization is statistically significant. From a functional point of view, this differential synaptic organization might indicate that lemniscal inputs have greater influence than spinal inputs in affecting the activity of thalamic neurons. Labeled spinothalamic terminals contact the same dendritic profile as do degenerating lemniscal terminals in about 10% of single sections. Because the present study did not include a complete reconstruction of ventrobasal complex neurons of the thalamus or even regions of dendritic arbors, the degree of convergence is likely to be significantly underestimated. These findings indicate that the anatomical basis exists for an interaction between nociceptive and non-nociceptive somesthetic systems at the level of single ventrobasal neurons of the thalamus of the rat.

Monosynaptic projections from the nucleus retroambiguus region to laryngeal motoneurons in the rhesus monkey.

Vocalization and straining-related activities require the activation of laryngeal muscles. The control of laryngeal muscles during these activities is thought to be mediated by a pathway from the periaqueductal gray via premotor neurons in the nucleus retroambiguus to laryngeal motoneurons in the nucleus ambiguus. However, direct contacts between the nucleus retroambiguus and laryngeal motoneurons have never been demonstrated anatomically. Moreover, data in primates about the nucleus retroambiguus-nucleus ambiguus pathway are lacking. Therefore, the present study examines the projection from the nucleus retroambiguus region to laryngeal motoneurons in the rhesus monkey at the light and electron microscopic levels. Injections with wheat germ agglutinin-horseradish peroxidase were made into the nucleus retroambiguus in five rhesus monkeys to anterogradely label fibers in the nucleus ambiguus. In two of these animals, the cricothyroid muscle was injected with cholera toxin subunit b to identify the motoneurons that supply it. The results show that the nucleus retroambiguus region most densely projects to the compact formation of the nucleus ambiguus, whereas cricothyroid motoneurons, which surround the compact formation, receive a moderate projection. The projections are bilateral, with a contralateral predominance. Ultrastructurally, anterogradely labeled terminal profiles from the nucleus retroambiguus contact cholera toxin subunit b-labeled dendrites of cricothyroid motoneurons. The terminal profiles contain primarily spherical vesicles and form asymmetrical contacts with cricothyroid motoneurons. This study demonstrates that the nucleus retroambiguus region projects to the nucleus ambiguus in the primate. Some of these projections include monosynaptic connections to laryngeal motoneurons. This pathway is important for the control of the vocal folds during vocalization and straining-related activities.

Projections from estrogen receptor-alpha immunoreactive neurons in the periaqueductal gray to the lateral medulla oblongata in the rhesus monkey.

The periaqueductal gray (PAG) contains numerous estrogen receptor-alpha immunoreactive (ER-alpha IR) neurons that are distributed in a species-specific way. These neurons might modulate different types of behavior that are mediated by the PAG such as active and passive coping responses, analgesia, and reproductive behavior. In primates, it is not known whether ER-alpha IR PAG neurons represent local interneurons and/or neurons that project to brainstem areas that control these behaviors. In this double labeling study, we asked whether ER-alpha IR neurons in the PAG of the rhesus monkey project to the nucleus retroambiguus (NRA), an area in the ventrolateral caudal medulla oblongata that is involved in expiration, vocalization, and reproductive behavior. Tracer was injected into the caudal lateral medulla oblongata to retrogradely label PAG neurons, and ER-alpha was visualized immunohistochemically. Although ER-alpha IR neurons and NRA-projection neurons were present at similar levels of the PAG, their distributions hardly overlapped. ER-alpha IR PAG neurons that project to the lateral caudal medulla represented less than 2% of ER-alpha IR PAG neurons. These double-labeled neurons were mainly located in the ipsilateral caudal PAG. The cluster of neurons in the medial part of the lateral PAG that projects specifically to the NRA-region did not contain double-labeled cells. The results indicate that only a few ER-alpha IR PAG neurons project to the NRA-region. This might be related to the modest effects of estrogen on mating-related behavior in primates compared most other mammalian species. Remaining ER-alpha IR PAG neurons might act locally on other PAG neurons, or they might represent neurons that project to other areas. Furthermore, the finding that the distributions of ER-alpha IR neurons and neurons that project to premotor neurons in the NRA-region scarcely overlap illustrates that the PAG in primates is very highly organized into anatomically distinct regions compared with other species.

Fine structure of the spinothalamic projections to the central lateral nucleus of the rat thalamus.

The fine structure of labelled spinothalamic terminals in the central lateral nucleus has been studied in the rat following injection of wheat germ agglutinin-horseradish peroxidase into the spinal cord. Myelinated axons gave rise to the labelled terminals, which were large profiles which contained round vesicles, numerous mitochondria, and formed asymmetrical contacts with large dendrites or dendritic protrusions. These profiles are similar to those described in other somatosensory thalamic nuclei, and in many other nuclei of the thalamus.

Reticularis thalami afferents to the ventrobasal complex of the rat thalamus: an electron microscope study.

Afferents from the nucleus reticularis thalami (RT) to the thalamic ventrobasal complex were studied in the rat by looking for degenerating terminals after selective neurotoxic lesion of RT using injections of kainic acid. Several lines of evidence are presented indicating that RT afferents terminate in the VB by F type (Gray type II) terminals and that F type terminals in the VB all depend of RT neurons.