Topographic organization of medial pulvinar connections with the prefrontal cortex in the rhesus monkey

The medial nucleus of the pulvinar complex (PM) has widespread connections with association cortex. We investigated the connections of the PM with the prefrontal cortex (PFC) in macaque monkeys, with tracers placed into the PM and the PFC, respectively. Injections of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) placed into the PM resulted in widespread anterograde terminal labeling in layers III and IV, and retrograde cellular labeling in layer VI of the PFC. Injections of tracers centered on the central/lateral PM resulted in labeling of dorsolateral and orbital regions, whereas injections centered on caudal, medial PM resulted in labeling of dorsomedial and medial PFC. Since injections of the PM included neighboring thalamic nuclei, retrograde tracers were placed into distinct cytoarchitectonic regions of the PFC and retrogradely labeled cells in the posterior thalamus were charted. The results of this series of tracer injections confirmed the results of thalamic injections. Injections placed into areas 8a, 12 (lateral and orbital), 45, 46 and 11, retrogradely labeled neurons in the central/lateral PM, while tracer injections placed into areas 9, 12 (lateral), 10 and 24, labeled medial PM. The connections of the PM with temporal, parietal, insular, and cingulate cortices were also examined. The central/lateral PM has reciprocal connections with posterior parietal areas 7a, 7ip, and 7b, insular cortex, caudal superior temporal sulcus (STS), caudal superior temporal gyrus (STG), and posterior cingulate, whereas medial PM is connected mainly with the anterior STS and STG, as well as the cingulate cortex and the amygdala. These connectional studies suggest that the central/ lateral and medial PM have divergent connections which may be the substrate for distinct functional circuits.     

Contralateral cortical projection to the mediodorsal thalamic nucleus: origin and synaptic organization in the rat

The origin of the corticothalamic projections to the contralateral mediodorsal nucleus, the collateralization of cortical fibers and their synaptic organization in the ipsi- and contralateral mediodorsal nuclei were investigated in adult rats with double retrograde fluorescent and anterograde tracing. After tracer injections in the mediodorsal nuclei on either side, neurons were retrogradely labeled in all the areas of the contralateral prefrontal cortex in which ipsilateral labeling was also observed. Contralateral corticothalamic cells accounted for 15% of the labeled neurons in the orbital and agranular insular areas, while their proportion was lower (3%) in the anterior cingulate cortex. Up to 70% of the contralateral cortical neurons were double labeled by bilateral injections in the mediodorsal nuclei. At the electron microscopic level, unilateral injections of biotinylated dextran-amine in the orbitofrontal cortex resulted in anterograde labeling of small terminals and a few large boutons in the ipsilateral mediodorsal nucleus, while only small boutons were identified contralaterally. The diameter of postsynaptic dendritic profiles contacted by labeled small cortical endings was significantly larger in the ipsilateral mediodorsal nucleus than contralaterally. These findings demonstrate that dense contralateral cortical projections to the mediodorsal nucleus derive from the orbital and agranular insular areas, and that crossed corticothalamic afferents are mostly formed by collaterals of the ipsilateral connections. Our observations also point out the heterogeneity of corticothalamic boutons in the rat mediodorsal nucleus and morphological differences in the synaptic organization of prefrontal fibers innervating the two sides, indicating that ipsilateral cortical afferents may be more proximally distributed than crossed cortical fibers on dendrites of mediodorsal neurons.     

Extensive divergence and convergence in the thalamocortical projection to monkey somatosensory cortex

This study examined the extent of thalamocortical divergence as a potential determinant of activity-dependent representational plasticity in area 3b of adult monkey somatosensory cortex. Single or paired injections of anterogradely transported tracers, of varying anteroposterior extent, were made horizontally from behind in defined parts of the body representation in the ventral posterior lateral (VPL) and/or ventral posterior medial (VPM) thalamic nuclei, and the distribution and density of labeled thalamocortical terminations were mapped in cortex. Injections of increasing size in any dimension of VPL or VPM resulted in increasing accumulation of labeled terminals within the region of projection, implying extensive convergence of individual axons. Anteroposteriorly elongated injections labeled mediolaterally extended but anteroposteriorly restricted zones in cortex. Dorsoventral placement of an injection in VPL or VPM determined anteroposterior location of labeling in cortex. Dual injections separated mediolaterally or dorsoventrally by approximately 1 mm, and in different parts of the thalamic body or head-face representation gave rise to labeled thalamocortical terminations that overlapped extensively. For injection sites at different anteroposterior levels in VPL or VPM, the area of cortical convergence was related to their extent of anteroposterior coincidence. Labeled terminations arising from injections in immediately adjacent parts of VPL and VPM did not overlap in cortex. The extent of thalamocortical divergence and convergence revealed by these experiments is greater than that predictable from labeling of single axons and is sufficiently great to account for representational plasticity that exceeds the 1.5 mm cortical "distance limit."     

Immunocytochemical localization and description of neurons expressing serotonin2 receptors in the rat brain

Serotonin2 receptors have been implicated in a variety of behavioral and physiological processes, as well as a number of neuropsychiatric disorders. To specify the brain regions and specific cell types possessing serotonin2 receptors, we conducted an immunocytochemical study of the rat brain using a polyclonal serotonin2 receptor antibody. Perfusion-fixed rat brain sections were processed for immunocytochemistry and reactivity was visualized using an immunoperoxidase reaction. Numerous small, round neurons were heavily labeled in the granular and periglomerular regions of the olfactory bulb. Heavy labeling of medium-sized multipolar and bipolar neurons was also seen in olfactory regions of the ventral forebrain, including the anterior olfactory nucleus and olfactory tubercle. Other regions of the basal forebrain exhibiting high levels of immunoreactivity were the nucleus accumbens, ventral pallidum, Islands of Calleja, fundus striatum and endopyriform nucleus. Immunoreactive neurons were also seen in the lateral amygdala. A dense band of small, round cells was stained in layer 2 of pyriform cortex. In neocortex, a very sparse and even distribution of bipolar and multipolar neurons was seen throughout layers II-VI. A much more faintly labeled population of oval cells was observed in the deep layer of retrosplenial and posterior cingulate cortex, and in the granular layer of somatosensory frontoparietal cortex. A moderate number of medium bipolar and multipolar cells were scattered throughout the neostriatum, and a moderate number of pyramidal and pyramidal-like cells were seen in the CA fields of the hippocampus. Diencephalic areas showing immunolabeling included the medial habenula and anterior pretectal nucleus, with less labeling in the ventral lateral geniculate. In the hindbrain, two dense populations of large multipolar cells were heavily labeled in the pedunculopontine and laterodorsal tegmental nuclei, with lesser labeling in the periaqueductal gray, superior colliculus, spinal trigeminal nucleus and nucleus of the solitary tract. Based on the distribution, localization and morphology of immunoreactive neurons in these regions, we hypothesize that subpopulations of serotonin2 containing cells may be GABAergic interneurons or cholinergic neurons. Further, the observed distribution suggests that the physiological effects of serotonin acting through serotonin2 receptors are mediated by a relatively small number of cells in the brain. These observations may have strong functional implications for the pharmacological treatment of certain neuropsychiatric disorders.     

Topographical distribution of NADPH-diaphorase activity in the central nervous system of the frog, Rana perezi

The distribution of NADPH-diaphorase (ND) activity was histochemically investigated in the brain of the frog Rana perezi. This technique provides a highly selective labeling of neurons and tracts. In the telencephalon, labeled cells are present in the olfactory bulb, pallial regions, septal area, nucleus of the diagonal band, striatum, and amygdala. Positive neurons surround the preoptic and infundibular recesses of the third ventricle. The magnocellular and suprachiasmatic hypothalamic nuclei contain stained cells. Numerous neurons are present in the anterior, lateral anterior, central, and lateral posteroventral thalamic nuclei. Positive terminal fields are organized in the same thalamic areas but most conspicuously in the visual recipient plexus of Bellonci, corpus geniculatum of the thalamus, and the superficial ventral thalamic nucleus. Labeled fibers and cell groups are observed in the pretectal area, the mesencephalic optic tectum, and the torus semicircularis. The nuclei of the mesencephalic tegmentum contain abundant labeled cells and a conspicuous cell population is localized medial and caudal to the isthmic nucleus. Numerous cells in the rhombencephalon are distributed in the octaval area, raphe nucleus, reticular nuclei, sensory trigeminal nuclei, nucleus of the solitary tract, and, at the obex levels, the dorsal column nucleus. Positive fibers are abundant in the superior olivary nucleus, the descending trigeminal, and the solitary tracts. In the spinal cord, a large population of intensely labeled neurons is present in all fields of the gray matter throughout its rostrocaudal extent. Several sensory pathways were heavily stained including part of the olfactory, visual, auditory, and somatosensory pathways. The distribution of ND-positive cells did not correspond to any single known neurotransmitter or neuroactive molecule system. In particular, abundant codistribution of ND and catecholamines is found in the anuran brain. Double labeling techniques have revealed restricted colocalization in the same neurons and only in the posterior tubercle and locus coeruleus. If ND is in amphibians a selective marker for neurons containing nitric oxide synthase, as generally proposed, with this method the neurons that may synthesize nitric oxide would be identified. This study provides evidence that nitric oxide may be involved in novel tasks, primarily related to forebrain functions, that are already present in amphibians.     

Monoclonal antibody Cat-301 selectively identifies a subset of nuclei in the cat's somatosensory thalamus

Recently it has been demonstrated that the monoclonal antibody Cat-301 is capable of identifying functionally related neurons in the mammalian visual thalamus. We have examined the possibility that this antibody might display a similar capacity in nonvisual thalamic areas. We demonstrate that in the cat's somatosensory thalamus the distribution of Cat-301-positive cells and neuropil is restricted to a subset of nuclei. These include the ventroposterior medial, ventroposterior lateral, and ventroposterior inferior nuclei. Staining with Cat-301 provides a clear visualisation of the entire somatotopic map within these nuclei. The somatosensory sector of the thalamic reticular nucleus and the perireticular nucleus, which may have a somatosensory sector, are also Cat-301-positive. In contrast, cells that do not express the Cat-301 antigen are located in the ventroposterior oralis nucleus, the ventroposterior shell region, the medial and lateral divisions of the posterior nuclear group, and the inner small cell region adjacent to the thalamic reticular nucleus. In comparison with previous physiological studies, cells that express the Cat-301 antigen most likely represent subpopulations in only a few of the somatic submodality-specific groups. These include cells in the small-field and Pacinian cutaneous-responsive groups, excluding cells in the wide-field cutaneous-, muscle-, joint-, and noxious-responsive groups. Taken together these findings indicate that monoclonal antibody Cat-301 is capable of selectively identifying neurons with distinct functional properties in the mammalian somatosensory thalamus.     

Organization of projections from the basomedial nucleus of the amygdala: a PHAL study in the rat

The organization of axonal projections from the basomedial nucleus of the amygdala (BMA) was examined with the Phaseolus vulgaris leucoagglutinin (PHAL) method in adult male rats. The anterior and posterior parts of the BMA, recognized on cytoarchitectonic grounds, display very different projection patterns. Within the amygdala, the anterior basomedial nucleus (BMAa) heavily innervates the central, medial, and anterior cortical nuclei. In contrast, the posterior basomedial nucleus (BMAp) sends a dense projection to the lateral nucleus, and to restricted parts of the central and medial nuclei. Extra-amygdalar projections from the BMA are divided into ascending and descending components. The former end in the cerebral cortex, striatum, and septum. The BMAa mainly innervates olfactory (piriform, transitional) and insular areas, whereas the BMAp also innervates inferior temporal (perirhinal, ectorhinal) and medial prefrontal (infralimbic, prelimbic) areas and the hippocampal formation. Within the striatum, the BMAa densely innervates the striatal fundus, whereas the nucleus accumbens receives a heavy input from the BMAp. Both parts of the BMA send massive projections to distinct regions of the bed nuclei of the stria terminalis. Descending projections from the BMA end primarily in the hypothalamus. The BMAa sends a major input to the lateral hypothalamic area, whereas the BMAp innervates the ventromedial nucleus particularly heavily. Injections were also placed in the anterior cortical nucleus (COAa), a cell group superficially adjacent to the BMAa. PHAL-labeled axons from this cell group mainly ascend into the amygdala and olfactory areas, and descend into the thalamus and lateral hypothalamic area. Based on connections, the COAa and BMAa are part of the same functional system. The results suggest that cytoarchitectonically distinct anterior and posterior parts of the BMA are also hodologically distinct and form parts of distinct anatomical circuits probably involved in mediating different behaviors (for example, feeding and social behaviors vs. emotion-related learning, respectively).     

Intrinsic GABAergic neurons in the rat central extended amygdala

The present study examined the distribution, morphology, and connections of gamma-aminobutyric acid-immunoreactive (GABA-IR) neurons in the three principal components of the central extended amygdala: the central amygdaloid nucleus, the bed nucleus of the stria terminalis (BNST) and the sublenticular substantia innominata. In the central nucleus, large numbers of GABA-IR neurons were identified in the lateral, lateral capsular, and ventral subdivisions, though in the medial subdivision, GABA-IR neurons were only present at very caudal levels. Combined immunocytochemistry-Golgi impregnation revealed that GABA-IR neurons in the lateral central nucleus were medium-sized spiny neurons that were morphologically similar to GABAergic neurons in the striatum. Injections of horseradish peroxidase into the bed nucleus of the stria terminalis labeled a major proportion of the GABA-IR neurons in the central nucleus. In the bed nucleus, the majority of GABA-IR neurons were located in the anterolateral subdivision, ventral part of the posterolateral subdivision and the parastrial subdivision. GABA-IR neurons in the anterolateral bed nucleus were of the typical medium-sized spiny type. Injections of horseradish peroxidase into the central nucleus labeled a few GABA-IR neurons in the posterior part of the anterolateral bed nucleus. GABA-IR neurons were identified in the sublenticular substantia innominata and medial shell of the nucleus accumbens and contributed to the continuum of GABA-IR extending from the central nucleus to the bed nucleus. Injections of horseradish peroxidase (HRP) into the central nucleus, but not the BNST, labeled a few GABA-IR neurons in the substantia innominata. The data point to GABA-IR neurons being a characteristic feature of the central extended amygdala and that GABA-IR neurons participate in the long intrinsic connections linking the major components of this structure. Since lesions of the stria terminalis and basolateral amygdaloid nucleus failed to deplete GABA-IR terminals in the central nucleus, the role of GABA in local and short intrinsic connections in the central extended amygdala is discussed. Further, physiological findings implicating the intrinsic GABAergic system of the central extended amygdala in the tonic inhibition of brainstem efferents are reviewed.     

An investigation of a possible direct projection from the medial nucleus of the cerebellum to the paraventricular nucleus of the hypothalamus in the rat: a study using retrograde WGA-HRP and Fluoro-Gold tracing techniques

Retrograde transport of lectin-conjugated horseradish peroxidase and Fluoro-Gold was used in an attempt to obtain data to confirm the existence, predicted from physiological studies, of a direct, monosynaptic projection from the medial nucleus of the cerebellum (MN) to the paraventricular nucleus of the hypothalamus (PVH) in the rat. Injections of these two tracers that included the PVH and surrounding diencephalic structures, or that in the case of Fluoro-Gold were localized to the PVH, resulted in retrograde neuronal labeling in widely separated nuclei known to project to the areas included in the injection sites. Thus, effective uptake and transport of both tracers occurred under the experimental conditions employed in this study. However, injections confined to the PVH and regions of the hypothalamus adjacent to it, or to the PVH alone, produced no retrograde neuronal labeling in the medial nucleus, indicating that the MN does not project directly to the PVH. Alternative explanations for the findings from physiological experiments were sought. The possibility that electrical stimulation of fibers of passage through the region of the MN might produce a monosynaptic response in the contralateral PVH was discarded, because retrogradely labeled neurons in nuclei such as the locus ceruleus and lateral parabrachial nucleus were distributed mainly ipsilateral to hypothalamic injection sites. However, tracer injections into the MN produced retrograde labeling of neurons in the same region of the lateral paragigantocellular nucleus (LPGi) in which labeled cells were found following tracers injections into the PVH. Axon collaterals of individual neurons in the LPGi might, therefore, project both to the MN and to the PVH. The possibility that such a circuit could, in the absence of a direct MN to PVH projection, provide the basis to explain the physiological findings is discussed.     

Differential projection patterns of superior and inferior collicular neurons onto posterior paralaminar nuclei of the thalamus surrounding the medial geniculate body in the rat

The thalamic nuclei at the medial border of the medial geniculate body (i.e. the suprageniculate nucleus, the medial division of the medial geniculate nucleus, the posterior intralaminar nucleus and the peripeduncular nucleus) which relay sensory information to the amygdala are thought to receive convergent input from multiple sites. In order to delineate the organization of these multimodal thalamic nuclei, the locations of superior and inferior collicular neurons projecting to these nuclei were studied by means of retrograde transport methods. Small injections of the tracer Miniruby were made into single paralaminar thalamic nuclei. Injections of Miniruby into the suprageniculate nucleus labelled predominantly neurons in the stratum opticum of the superior colliculus, whereas injections into the medial division of the medial geniculate body, the posterior intralaminar nucleus and the peripeduncular nucleus labelled predominantly neurons in the deep layers of the superior colliculus. These injections also labelled neurons in the inferior colliculus. The majority of retrogradely labelled neurons were found in the external nucleus of the inferior colliculus and here predominantly in layer 2. Injections focused onto the medial division of the medial geniculate body additionally labelled magnocellular neurons in layer 3 of the external nucleus and a few neurons in the central nucleus. More ventrally located injections, focused onto the posterior intralaminar and peripeduncular nucleus, almost exclusively labelled neurons in layer 1 of the external nucleus and the dorsal part of the dorsal nucleus. After injections into the suprageniculate nucleus, only neurons in layer 2 were found. Neurons in the central nucleus of the inferior colliculus were only found after injections that involved the medial division of the medial geniculate body. The present results suggest that, despite a considerable degree of convergence in this thalamic region, each of these thalamic nuclei receives a unique pattern of projections from the superior and inferior colliculi. It appears that the thalamic nuclei may be concerned mainly, but not exclusively, with a single sensory modality, and give rise to parallel multimodal and unimodal pathways to the amygdala.     

Somatovisceral projections from spinal cord and dorsal column nuclei to the thalamus in hedgehog tenrecs

In order first to overcome the difficulties in understanding the increasing amount of information available regarding the mammalian somatosensory thalamus, and then to correlate the findings among different species and integrate them into a general concept of thalamic organization, the present study investigated the spinothalamic and medial lemniscal projections in Madagascan hedgehog tenrecs (Echinops telfairi and Setifer setosus). Tracer substances were injected into the dorsal column nuclei and into spinal segments at various levels; additional injections were made into the inferior colliculus. The ascending somesthetic projections were to predominantly contralateral posterolateral target areas, and were almost mirror-like on both sides to intralaminar and medial thalamic nuclei. The densest and most extensive projections, originating mainly from the high cervical spinal cord and the dorsal column nuclei, reached the posterolateral thalamus caudal to the lateral geniculate nucleus. This region was difficult to subdivide cytoarchitecturally; nevertheless, on the basis of its labeling pattern, several subdivisions could be described and preliminary named. Some of them compared tentatively with the internal portion of the medial geniculate nucleus (GM) and the ventral posterior nuclear complex (VPC) in more differentiated mammals. The most prominent subdivision, however, located subjacent to the lateral surface of the brainstem, was shown to receive additional fibers from the inferior colliculus. This region might be considered a further subdivision of GM, VPC, a perigeniculate area, and/or a region of its own not comparable at present, with thalamic regions in other mammals. On the other hand, it may also be a remnant of the hypothetical, diffuse multimodal region from which GM and VPC have possibly evolved.(ABSTRACT TRUNCATED AT 250 WORDS)     

Transneuronal pathways to the vestibulocerebellum

The alpha-herpes virus (pseudorabies, PRV) was used to observe central nervous system (CNS) pathways associated with the vestibulocerebellar system. Retrograde transneuronal migration of alpha-herpes virions from specific lobules of the gerbil and rat vestibulo-cerebellar cortex was detected immunohistochemically. Using a time series analysis, progression of infection along polyneuronal cerebellar afferent pathways was examined. Pressure injections of > 20 nanoliters of a 10(8) plaque forming units (pfu) per ml solution of virus were sufficient to initiate an infectious locus which resulted in labeled neurons in the inferior olivary subnuclei, vestibular nuclei, and their afferent cell groups in a progressive temporal fashion and in growing complexity with increasing incubation time. We show that climbing fibers and some other cerebellar afferent fibers transported the virus retrogradely from the cerebellum within 24 hours. One to three days after cerebellar infection discrete cell groups were labeled and appropriate laterality within crossed projections was preserved. Subsequent nuclei labeled with PRV after infection of the flocculus/paraflocculus, or nodulus/uvula, included the following: vestibular (e.g., z) and inferior olivary nuclei (e.g., dorsal cap), accessory oculomotor (e.g., Darkschewitsch n.) and accessory optic related nuclei, (e.g., the nucleus of the optic tract, and the medial terminal nucleus); noradrenergic, raphe, and reticular cell groups (e.g., locus coeruleus, dorsal raphe, raphe pontis, and the lateral reticular tract); other vestibulocerebellum sites, the periaqueductal gray, substantia nigra, hippocampus, thalamus and hypothalamus, amygdala, septal nuclei, and the frontal, cingulate, entorhinal, perirhinal, and insular cortices. However, there were differences in the resulting labeling between infection in either region. Double-labeling experiments revealed that vestibular efferent neurons are located adjacent to, but are not included among, flocculus-projecting supragenual neurons. PRV transport from the vestibular labyrinth and cervical muscles also resulted in CNS infections. Virus propagation in situ provides specific connectivity information based on the functional transport across synapses. The findings support and extend anatomical data regarding vestibulo-olivo-cerebellar pathways.     

Organization of neural inputs to the suprachiasmatic nucleus in the rat

The circadian timing of the suprachiasmatic nucleus (SCN) is modulated by its neural inputs. In the present study, we examine the organization of the neural inputs to the rat SCN using both retrograde and anterograde tracing methods. After Fluoro-Gold injections into the SCN, retrogradely labeled neurons are present in a number of brain areas, including the infralimbic cortex, the lateral septum, the medial preoptic area, the subfornical organ, the paraventricular thalamus, the subparaventricular zone, the ventromedial hypothalamic nucleus, the posterior hypothalamic area, the intergeniculate leaflet, the olivary pretectal nucleus, the ventral subiculum, and the median raphe nuclei. In the anterograde tracing experiments, we observe three patterns of afferent termination within the SCN that correspond to the photic/raphe, limbic/hypothalamic, and thalamic inputs. The median raphe projection to the SCN terminates densely within the ventral subdivision and sparsely within the dorsal subdivision. Similarly, areas that receive photic input, such as the retina, the intergeniculate leaflet, and the pretectal area, densely innervate the ventral SCN but provide only minor innervation of the dorsal SCN. A complementary pattern of axonal labeling, with labeled fibers concentrated in the dorsal SCN, is observed after anterograde tracer injections into the hypothalamus and into limbic areas, such as the ventral subiculum and infralimbic cortex. A third, less common pattern of labeling, exemplified by the paraventricular thalamic afferents, consists of diffuse axonal labeling throughout the SCN. Our results show that the SCN afferent connections are topographically organized. These hodological differences may reflect a functional heterogeneity within the SCN.     

Cortical afferents of the perirhinal, postrhinal, and entorhinal cortices of the rat

We have divided the cortical regions surrounding the rat hippocampus into three cytoarchitectonically discrete cortical regions, the perirhinal, the postrhinal, and the entorhinal cortices. These regions appear to be homologous to the monkey perirhinal, parahippocampal, and entorhinal cortices, respectively. The origin of cortical afferents to these regions is well-documented in the monkey but less is known about them in the rat. The present study investigated the origins of cortical input to the rat perirhinal (areas 35 and 36) and postrhinal cortices and the lateral and medial subdivisions of the entorhinal cortex (LEA and MEA) by placing injections of retrograde tracers at several locations within each region. For each experiment, the total numbers of retrogradely labeled cells (and cell densities) were estimated for 34 cortical regions. We found that the complement of cortical inputs differs for each of the five regions. Area 35 receives its heaviest input from entorhinal, piriform, and insular areas. Area 36 receives its heaviest projections from other temporal cortical regions such as ventral temporal association cortex. Area 36 also receives substantial input from insular and entorhinal areas. Whereas area 36 receives similar magnitudes of input from cortices subserving all sensory modalities, the heaviest projections to the postrhinal cortex originate in visual associational cortex and visuospatial areas such as the posterior parietal cortex. The cortical projections to the LEA are heavier than to the MEA and differ in origin. The LEA is primarily innervated by the perirhinal, insular, piriform, and postrhinal cortices. The MEA is primarily innervated by the piriform and postrhinal cortices, but also receives minor projections from retrosplenial, posterior parietal, and visual association areas.     

Neurons located in the trigeminal sensory complex and the lateral pontine tegmentum project to the oculomotor nucleus in the rabbit

Neurons located in the trigeminal sensory complex (TSC) and the lateral pontine tegmentum (LPT) have been reported to project to both the accessory abducens and the facial nuclei, which innervate the retractor bulbi and orbicularis oculi muscles respectively, in order to control the nictitating membrane (NM) and eyelid defensive reflex. Since muscles innervated by the oculomotor nucleus (OCM) also appear to be involved in this reflex, retrograde and anterograde tracers were used in this study to determine whether there are projections from the TSC and LPT to the OCM in the rabbit. Injections of horseradish peroxidase (HRP) in the OCM nucleus labeled neurons in the LPT surrounding the trigeminal motor nucleus dorsally, laterally and ventrally. Only a few scattered neurons were found in the principal and spinal trigeminal nuclei. Injection of biocytin in the LPT area containing most of the HRP-labeled neurons caused anterograde labeling of fibers that crossed the midline and ascended just dorsal to the contralateral medial lemniscus. A proportion of these fibers coursed in a dorsal direction to enter and terminate within the OCM contralateral to the injection site. The location of the motoneuronal groups innervating the different extraocular muscles was studied by retrograde transport of HRP, and compared with the distribution of biocytin-labeled terminals. It was found that the terminals were located in the superior rectus and the levator palpebrae zone of the nucleus. We discuss the functional significance of this projection for the eyelid and NM response.     

Oestradiol-releasing vaginal ring for treatment of postmenopausal urogenital atrophy

In order to investigate the possible routes linking the thalamus in the two sides of the brain, the connections of the reticular nucleus (RT), the major component of the ventral thalamus, with contralateral dorsal thalamic nuclei were systematically investigated in the adult rat. This study was performed with several tract-tracing techniques: single and double retrograde labeling with fluorescent tracers, and anterograde tracing with biocytin. Retrograde tracing was also combined with immunocytochemistry to provide additional criteria for the identification of labeled RT neurons. The data obtained with the retrograde transport of one fluorescent tracer showed that RT neurons project to contralateral dorsal thalamic domains. In particular, retrograde labeling findings indicated that the anterior intralaminar nuclei, as well as the ventromedial (VM) nucleus, are preferential targets of the contralateral RT projections. Commissural neurons were concentrated in two portions of RT: its rostral part, including the rostral pole, which projects to the contralateral central lateral (CL) and paracentral (Pc) nuclei, and the ventromedial sector of the middle third of RT, which projects to the contralateral VM and posterior part of CL and Pc. The double retrograde labeling study of the bilateral RT-intralaminar connection indicated that at least part of the commissural RT cells bifurcate bilaterally to symmetrical portions of the anterior intralaminar nuclei. The targets of the RT commissural system inferred from the retrograde labeling data were largely confirmed by anterograde tracing. Moreover, it was shown that RT fibers cross the midline in the intrathalamic commissure. The present data demonstrate that bilateral RT connections with the dorsal thalamus provide a channel for interthalamic crosstalk. Through these bilateral connections with thalamic VM and intralaminar neurons, RT could influence the activity of wide territories of the cerebral cortex and basal ganglia of both hemispheres.     

Mapping of angiotensin AT1 receptors in the rat limbic system

The AT1 receptor is one of the two receptor subtypes able to bind angiotensin II. In the present study, immunohistochemical examination of the distribution of the AT1 receptor in several limbic structures of female rats has been done, revealing new aspects of the distribution of AT1-positive cells. The presence of AT1 receptor expressing cells in the hippocampus and the amygdala is described, but their distribution in these regions has not been examined in a detailed way. We found some notable differences in the distribution of these cells: in female rats, we detected high amounts of labeled cells in the hippocampus, the entorhinal cortex and piriform cortex. In somewhat lower amounts, stained cells could be found in several nuclei of the amygdala (in the basomedial, basolateral, lateral, central and medial nucleus of the amygdala, in the amygdalopiriform transition area and in the amygdalohippocampal transition area as well as in the bed nucleus of the stria terminalis).     

Diversity of connections of the temporal neocortex with amygdaloid nuclei in the dog (Canis familiaris)

Reciprocal connections of amygdaloid nuclei with the temporal neocortex in the dog were investigated. Injections of fluorescent tracers and BDA into particular temporal areas were made in eleven dogs. The topographical arrangement of connections and variations in their density differentiate the temporal neocortex in the dog into a few regions. Among them, the cortex involving the anterior part of the ectosylvian gyrus did not send any amygdalopetal projection. The middle ectosylvian, dorsal zone of the posterior ectosylvian and the anterior part of the Sylvian gyrus were weakly connected with the amygdala. The cortical region involving the ventral zone of the posterior ectosylvian and composite posterior areas, as well as posterior Sylvian gyrus, was characterized by profuse connections with the amygdaloid complex. Cortico-amygdaloid connections originate in the wide cortical area of the auditory cortex of the middle and dorsal part of the posterior ectosylvian gyrus as well as in the auditory association cortex located in the ventral ectosylvian, composite posterior and posterior Sylvian gyri. The connections showed a dorso-ventral gradient of increasing density, in the direction of association fields. The most substantial projection taking rise from the ectosylvian posterior and posterior composite gyri terminated preferentially in the pericapsular sector of the lateral amygdaloid nucleus and, to a lesser degree, in its medial sector. Terminals of connections originating in the Sylvian gyrus occupied preferentially the intermediate part of the lateral nucleus, slightly more medially than that from the ectosylvian and posterior composite areas. Additionally, axonal terminals derived from the composite posterior and Sylvian posterior areas were observed in the basal parvocellular and magnocellular nuclei. Neocortical projections were reciprocated by amygdalofugal connections with two exceptions: the basal magnocellular nucleus was distinguished by a substantial amygdalofugal projection to the temporal neocortex focused on the dorsal Sylvian gyrus, and the central nucleus of the amygdala, in contrast, received an exclusively corticofugal projection.     

Recognition and management of childhood cardiac arrhythmias

Axonal connections between the amygdala and the hypothalamic paraventricular nucleus were examined by combined anterograde-retrograde tract tracing. Iontophoretic injections of the retrograde tracer Fluorogold were placed in the paraventricular nucleus, and the anterograde tracer PHA-L in the ipsilateral central or medial amygdaloid nuclei. Single and double-label immunohistochemistry were used to detect tracers. Single label anterograde and retrograde tracing suggest limited evidence for direct connections between the central or medial amygdala and the paraventricular nucleus. In general, scattered PHA-L-positive terminals were seen in autonomic subdivisions of the paraventricular nucleus (lateral parvocellular, dorsal parvocellular and ventral medial parvocellular subnuclei) following central or medial amygdaloid nucleus injection. Double-label studies indicate that central and medial amygdaloid nucleus efferents contact paraventricular nucleus-projecting cells in several forebrain nuclei. In the case of central nucleus injections, PHA-L positive fibers occasionally contacted Fluorogold-labeled neurons in the anteromedial, ventromedial and preoptic subnuclei of the bed nucleus of the stria terminalis. Overall, such contacts were quite rare, and did not occur in the bed nucleus of the stria terminalis regions showing greatest innervation by the central amygdaloid nucleus. In contrast, medial amygdala injections resulted in a significantly greater overlap of PHA-L labeling and Fluorogold-labeled neurons, with axosomatic appositions observed in medial divisions of the bed nucleus of the stria terminalis, anterior hypothalamic area and preoptic area. The results provide anatomical evidence that a substantial proportion of amygdaloid connections with hypophysiotrophic paraventricular nucleus neurons are likely multisynaptic, relaying in different subregions of the bed nucleus of the stria terminalis and hypothalamus.     

Visceral afferent pathways to the thalamus and olfactory tubercle: behavioral implications

The goal of this study was to support the hypothesis that visceral signals may integrate and influence behavior by way of direct pathways from the nucleus tractus solitarii (NTS) to the olfactory tubercle and the midline/intralaminar thalamus. An anterograde tracer, biotinylated dextran amine (BDA) was iontophoresed bilaterally into the caudal NTS to optimize terminal labeling. NTS-cortical projections traversed both limbs of the diagonal bands providing heavy innervation, and terminated lightly within layer 3 of the olfactory tubercle. NTS-thalamic projections terminated within anterior and, as previously shown, posterior divisions of nucleus paraventricularis thalami and avoided the adjoining mediodorsal thalamic nucleus. Heretofore unrecognized projections were traced to the parafascicular and reuniens thalamic nuclei, and the peripeduncular nucleus. Control experiments identified the nucleus gracilis as the principal source of ascending projections to ventroposterior lateral, posterior and intralaminar thalamic nuclei. Our data corroborate the supposition that olfactory signals may integrate with visceral stimuli in the striatal compartment of olfactory tubercle. NTS projections encompass thalamic nuclei that project topographically to the prefrontal cortex, hippocampus and ventral (limbic) striatum, regions activated by visceral stimulation. Structural data support the idea that compartments of the non-discriminative thalamus may contribute to perception and behavioral responses to visceral stimulation.