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PubMed1993

Spinal neurogenesis and axon projection: a correlative study in the rat.

The purpose of the present study was to determine the relationship between the duration of a spinal neuron's neurogenic period and the length of its axon or level of projection. Spinal segment L1 was chosen for examination and neurons were divided into four projection groups: 1) supraspinal projection (SSp), 2) long ascending propriospinal (LAPr), 3) short ascending propriospinal (SAPr), and 4) descending propriospinal (DPr). To determine the duration of the neurogenic period for each group, 3H-thymidine was administered to fetal rats during the proliferative period for spinal neuroblasts on one of embryonic (E) days E13 through E16. Between 50 and 100 days after birth neurons in each group were labeled with the retrograde fluorescent tracer Fluoro-Gold. To demonstrate nerve cells with SSp projections, spinal cords were hemisected at spinal segment C3 in one group of animals and Fluoro-Gold was applied to the sectioned surface of the cord. Three additional sets of animals were used to label nerve cells with LAPr, SAPr, and DPr projections by injecting Fluoro-Gold into the gray matter at spinal segments C6, T12, and L5, respectively. Neurons labeled with both Fluoro-Gold and 3H-thymidine and neurons labeled with Fluoro-Gold alone in each animal in each group were counted and the data statistically analyzed. Results showed that within each spinal lamina neurons with different projections were generated, i.e., completed cell division, at significantly different rates. Neurons with the longest axons, those with SSP projections, were generated first. These were followed by those with LAPr projections, and finally those with SAPr and DPr projections. In most laminate there was no significant difference between the neurogenic periods of rostrally projecting short propriospinal (SAPr) neurons versus caudally projecting short propriospinal (DPr) neurons. It was concluded that the duration of the neurogenic period for a given group of neurons within each spinal lamina is inversely related to the distance between the nerve cell and its projection site regardless of the direction of its projection.

Animals↗
PubMed1993

Neuronal projections to the medial preoptic area of the sheep, with special reference to monoaminergic afferents: immunohistochemical and retrograde tract tracing studies.

The preoptic area contains most of the luteinizing hormone releasing hormone immunoreactive neurons and numerous monoaminergic afferents whose cell origins are unknown in sheep. Using tract tracing methods with a specific retrograde fluorescent tracer, fluorogold, we examined the cells of origin of afferents to the medial preoptic area in sheep. Among the retrogradely labeled neurons, immunohistochemistry for tyrosine hydroxylase, dopamine-beta-hydroxylase, phenylethanolamine N-methyltransferase, and serotonin was used to characterize catecholamine and serotonin fluorogold labeled neurons. Most of the afferents came from the ipsilateral side to the injection site. It was observed that the medial preoptic area received major inputs from the diagonal band of Broca, the lateral septum, the thalamic paraventricular nucleus, the lateral hypothalamus, the area dorsolateral to the third ventricle, the perimamillary area, the amygdala, and the ventral part of the hippocampus. Other numerous, scattered, retrogradely labeled neurons were observed in the ventral part of the preoptic area, the vascular organ of the lamina terminalis, the ventromedial part of the hypothalamus, the periventricular area, the area lateral to the interpeduncular nucleus, and the dorsal vagal complex. Noradrenergic afferents came from the complex of the locus coeruleus (A6/A7 groups) and from the ventro-lateral medulla (group A1). However, dopaminergic and adrenergic neuronal groups retrogradely labeled with fluorogold were not observed. Serotoninergic fluorogold labeled neurons belonged to the medial raphe nucleus (B8, B5) and to the serotoninergic group situated lateral to the interpeduncular nucleus (S4). In the light of these anatomical data we hypothesize that these afferents have a role in the regulation of several functions of the preoptic area, particularly those related to reproduction. Accordingly these afferents could be involved in the control of luteinizing hormone releasing hormone (LHRH) pulsatility or of preovulatory LHRH surge.

Animals↗
PubMed1993

Aminergic and cholinergic afferents to REM sleep induction regions of the pontine reticular formation in the rat.

Microinjection of cholinergic agonists in a dorsolateral part of the mesopontine tegmentum has been shown to induce a rapid eye movement (REM) sleep-like state. Physiological evidence indicates that not only acetylcholine but also various amine transmitters, including those implicated in behavioral state regulation, affect neuronal activity in this region of the pontine reticular formation. In the present study, sources of select aminergic and cholinergic inputs to this REM sleep induction zone were identified and quantitatively analyzed by using fluorescence retrograde tracing combined with immunofluorescence in the rat. In addition to previously demonstrated cholinergic projections from the pedunculopontine and laterodorsal tegmental nuclei, the REM sleep induction zone received various aminergic inputs that originated in widely distributed regions of the brainstem and hypothalamus. Serotoninergic afferents represented a mean of 44% of all aminergic/cholinergic source neurons projecting to the REM sleep induction zone, which was comparable to the mean percentage of 39% represented by cholinergic afferent neurons. The serotoninergic afferents originated from the raphe nuclei at all brainstem levels, with heavier projections from the pontine than from the medullary raphe nuclei. Unexpectedly, an additional major serotoninergic input was provided by serotoninergic neurons in the nucleus prosupralemniscus (B9). Noradrenergic afferent neurons represented a mean of 14% of all aminergic/cholinergic source neurons, which was only about one-third of the mean percentage of either cholinergic or serotoninergic source neurons. These noradrenergic projection neurons were located not only in the locus ceruleus (8%) but also in the lateral tegmentum, including the A5 (4%) and A7 (2%) cell groups. Histaminergic neurons in the tuberomammillary hypothalamic nucleus represented a minor group of afferent neurons (3%), and a still smaller input came from adrenergic C1 neurons. The pattern of these transmitter-specific afferent connections appeared to be similar regardless of the longitudinal level within the REM sleep induction zone. The present results are consistent with previous behavioral and physiological evidence for a role of the pontine REM sleep induction zone in triggering REM sleep. The regulation of REM sleep induction would be best understood in terms of a state-dependent interplay of cholinergic, serotoninergic, and other inputs all acting convergently upon neurons in the REM sleep-inducing region of the pontine reticular formation.

Afferent Pathways↗
PubMed1993

Ventral subicular interaction with the hypothalamic paraventricular nucleus: evidence for a relay in the bed nucleus of the stria terminalis.

The axonal projections of the ventral subiculum to the bed nucleus of the stria terminalis (BST) were examined in the rat with the anterograde neuronal tracer Phaseolus vulgaris-leucoagglutinin (PHA-L). Axons originating in the ventral subiculum coursed to the BST through either the fimbria-fornix, or a pathway involving the stria terminalis via the amygdala. Ventral subicular axons gave rise to dense terminal networks that were preferentially distributed in medial and ventral subregions of the BST. The distribution of subicular fibers and terminals was examined in relation to BST neurons that project to the hypothalamic paraventricular nucleus (PVN). In these cases, discrete iontophoretic injections of the retrograde tracer Fluoro-gold were made in the PVN, with PHA-L delivered to the ipsilateral ventral subiculum. An immunocytochemical double-labeling protocol was then employed for the simultaneous detection of PHA-L and Fluoro-gold, and provided light microscopic evidence for subicular input to PVN-projecting cells located within the BST. In a second series of experiments, the gamma-amino butyric acid (GABA)ergic nature of the BST was examined by in situ hybridization histochemistry for detection of transcripts encoding GAD67 mRNA. The studies revealed that a high proportion of BST neurons express GAD67 transcripts. Also, experiments combining Fluoro-gold tracing with GAD67 in situ hybridization suggested that a proportion of PVN-projecting neurons in the BST are GABAergic. Taken together, the results of these sets of studies suggest that the inhibitory influences of the hippocampus on the PVN might be relayed through specific portions of the BST. These findings may have important implications for our understanding of the neural regulation of the hypothalamic-pituitary-adrenal axis.

Amygdala↗
PubMed1993

Innervation of the amygdaloid complex by catecholaminergic cell groups of the ventrolateral medulla.

The projections to the amygdaloid complex (AMG), originating in the catecholaminergic cell groups of the ventrolateral medulla (VLM), were studied in the rat by using either the retrograde tracer fluoro-gold (FG) or the anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L) in combination with tyrosine hydroxylase (TH) and/or phenylethanolamine-N-methyltransferase (PNMT) immunohistochemistry. In the first series of experiments, injections of FG were made into regions of the central nucleus of the amygdala (ACe) where dense TH and PNMT immunoreactivity was previously observed, and then sections of the brainstem were processed for TH and PNMT immunoreactivity. FG retrogradely labelled neuronal cell bodies were observed throughout the rostrocaudal extent of VLM, bilaterally, with a contralateral predominance. Approximately 44% of the FG labelled cell bodies in VLM were also immunoreactive to the catecholamine biosynthetic enzymes TH and/or PNMT. Most of these catecholaminergic neurons were part of the A1 noradrenergic cell group in the caudal VLM and to a lesser extent part of the C1 adrenergic cell group in the rostral VLM. In the second series of experiments, PHA-L was iontophoresed into VLM at different rostrocaudal levels where in the previous series of experiments FG retrogradely labelled cell bodies were observed. Transverse sections of the forebrain and brainstem were then processed for the demonstration of PHA-L and either TH or PNMT immunoreactivity in cell bodies, axons, and presumptive axon terminals. PHA-L injection sites within either the caudal or rostral VLM resulted in labelled axons and terminal bouton-like swellings primarily in the contralateral AMG and to a lesser extent in the ipsilateral AMG. The ACe was observed to receive the greatest innervation from either VLM site. Additionally, PHA-L labelled fibers and presumptive terminal boutons were observed within the intercalated, medial, basomedial, and basolateral nuclei of the AMG. Most of the PHA-L labelled fibers and presumptive terminal boutons in the AMG after a caudal VLM (A1 region) injection also displayed TH immunoreactivity, whereas after a PHA-L injection into the rostral VLM (C1 region) all of the labelled axons and axon terminals in the AMG also were immunoreactive to PNMT. These data demonstrate that catecholaminergic neurons in A1 and C1 regions of VLM innervate the AMG and suggest that these VLM neurons may be involved in relaying afferent information directly to the AMG which influences the activity of AMG neurons controlling autonomic, endocrine, and behavioural functions.

Afferent Pathways↗
PubMed1993

Choline acetyltransferase immunoreactive neurons innervating labyrinthine and lateral line sense organs in amphibians.

The goal of the present study was to investigate aspects of the central organization of the neurons belonging to the octavolateralis efferent system of amphibians. The perikarya of three genera, Pleurodeles, Xenopus, and Discoglossus, were located in the brainstem by applying retrograde tracers to the appropriate cranial nerves and choline acetyltransferase immunohistochemistry was used to identify cholinergic neurons. The efferent neurons supplying lateral line (Pleurodeles, Xenopus) and labyrinthine (Pleurodeles, Xenopus, and Discoglossus) end organs were found to intermingle in a single octavolateralis efferent nucleus. The neurons lie bilateral to the labelled nerves in Pleurodeles and ipsilateral in Xenopus and Discoglossus. Separate labelling of the anterior and posterior octavus rami provided no evidence for distinct groupings of efferent neurons that could be associated with auditory and vestibular end organs. In all three species many if not all octavolateral efferent neurons displayed immunoreactivity for choline acetyltransferase. They could be distinguished from the cholinergic facial motoneurons, with which they sometimes intermingle, on the basis of either their distinctive size and shape (Pleurodeles, Xenopus) or their location (Discoglossus). Double labelling in Xenopus confirmed the cholinergic nature of the efferent neurons.

Animals↗
PubMed1993

Organization of the hamster paraventricular hypothalamic nucleus.

The hamster periventricular hypothalamic area has been the focus of functional research concerning photoperiodic time measurement. These studies have relied upon the extensive analysis of rat paraventricular nucleus because there has been a general absence of anatomical description in the hamster. The present work sought to remedy this problem by investigating the structure of the hamster paraventricular nucleus with respect to the localization of cells immunoreactive to vasopressin, oxytocin, or corticotropin-releasing factor and of cells projecting to the spinal cord or to vascular sites outside the blood-brain barrier. The hamster paraventricular nucleus includes the medial, lateral, and posterior magnocellular divisions, the main parvicellular division, as well as the periventricular area and dorsal cap, which are also parvicellular. The magnocellular divisions are characterized by many large neurons immunoreactive to oxytocin and vasopressin, which are generally absent from the parvicellular divisions. In contrast, corticotrophin-releasing hormone-immunoreactive cells are plentiful in most of the parvicellular areas. Spinally projecting cells are found in two rostral areas, one dorsally and a second, more ventral area. More caudally, the two regions merge within the posterior magnocellular division. Cells of the ventral group are frequently immunoreactive for one of the three peptides. Cells identified by peripheral injection of retrograde label are found in the rostral magnocellular divisions but not in the caudal posterior magnocellular division. Areas in which these cells also contain peptide are also described. The features of the hamster paraventricular nucleus are compared to those in the rat and apparent species differences are discussed.

Animals↗
PubMed1993

Origin of serotoninergic afferents to the hypoglossal nucleus in the rat.

The hypoglossal nucleus contains serotonin and several different serotonin receptors, and serotonin is present in fibers and terminals contacting hypoglossal motoneurons. Serotonin alters the excitability of hypoglossal motoneurons, and may influence hypoglossal motoneuron activity in a variety of physiological processes. Since the hypoglossal nucleus contains no serotoninergic somata, the present study sought to identify the sources of serotoninergic afferents to the hypoglossal nucleus. Fluorogold was injected into the hypoglossal nucleus and serotoninergic immunofluorescence was utilized in a dual-fluorescence technique to identify the sources of serotoninergic afferents to the hypoglossal nucleus. The results demonstrate that most serotoninergic afferents to the hypoglossal nucleus originate from the nuclei raphe pallidus and obscurus, while fewer originate from the nucleus raphe magnus and the parapyramidal region. Other regions of the medial tegmental field and the pons that contain both serotoninergic neurons and neuronal afferents to the hypoglossal nucleus contain no double-labeled neurons.

Animals↗
PubMed1993

Estrogen receptor-immunoreactive forebrain neurons project to the ventrolateral hypothalamus in female guinea pigs.

In rodents, the facilitation of sexual receptivity by estradiol and progesterone is suspected to be mediated by a network of neurons containing estrogen and progestin receptors. In female guinea pigs, this network would include estrogen receptor-immunoreactive (ER-ir) neurons located within the rostro-ventral ventrolateral hypothalamus (r-vVLH). This hypothesis predicts that a proportion of the neurons projecting to the r-vVLH contains estrogen receptors. This prediction was tested through retrograde tracing combined with immunocytochemistry for estrogen receptors. Retrogradely labelled neurons were particularly abundant within the medial preoptic nucleus (MPN), bed nucleus of the stria terminalis (BST), anterior hypothalamus, amygdala, and lateral parabrachial nucleus. As predicted by the hypothesis, retrogradely labelled neurons were mostly observed in estrogen receptor-rich areas. Retrogradely labelled neurons also containing estrogen receptor-immunoreactivity (ER-IR) were primarily found within the MPN, BST, and amygdala. However, a majority of retrogradely labelled neurons did not contain ER-IR. As the preoptic area and the r-vVLH are both responsive to estradiol in the facilitation of sexual receptivity by progesterone, these data are consistent with the hypothesis tested. However, our data also suggest that the network of neurons controlling sexual receptivity may include elements not directly sensitive to estradiol. Finally, the location of retrogradely labelled neurons is discussed with respect to the stimuli provided to the r-vVLH in the context of sexual receptivity facilitated by estradiol and progesterone.

Animals↗
PubMed1993

Localization of cholinergic neurons in the forebrain and brainstem that project to the suprachiasmatic nucleus of the hypothalamus in rat.

In mammals, the suprachiasmatic nucleus is responsible for the generation of most circadian rhythms and their entrainment to environmental cues. Cholinergic agents can alter circadian rhythm phase, and fibres immunoreactive for choline acetyltransferase, the biosynthetic enzyme for acetylcholine, are present in the suprachiasmatic nucleus. Since there are no cholinergic somata in the suprachiasmatic nucleus, these fibres must represent the terminals of cholinergic neurons whose cell bodies are located elsewhere in the brain. This study was aimed at locating the cholinergic neurons that project to the suprachiasmatic nucleus by retrograde and anterograde tract-tracing and immunohistochemistry for choline acetyltransferase in the rat. After injection of fluorogold, a retrograde tracer, into the suprachiasmatic nucleus, retrogradely labelled neurons that were immunopositive for choline acetyltransferase were located throughout the rostrocaudal extent of the cholinergic basal nuclear complex, with highest densities in the substantia innominata and the nucleus basalis magnocellularis. A few cells were also located in the medial septum and in the vertical and horizontal limbs of the diagonal band of Broca. In the brainstem, double-labelled neurons were located in the laterodorsal tegmental nucleus, pedunculopontine tegmental nucleus and the parabigeminal nucleus. Injections of the anterograde tracer biocytin in these three brainstem nuclei resulted in fibre labelling in the suprachiasmatic nucleus, consistent with the retrograde findings. No clearly double-labelled cells were located in the retina. These results suggest that the suprachiasmatic nucleus receives cholinergic afferents from both the basal forebrain and mesopontine tegmentum which may mediate cholinergic effects on circadian rhythms.

Animals↗
PubMed1993

Retrograde labeling of neurons in the spinal cord that project directly to the amygdala or the orbital cortex in the rat.

The amygdala and orbital cortex are thought to play an important role in the regulation of autonomic functions, hormonal secretion, and behavioral expression in response to sensory stimulation. The responsiveness of neurons in these regions to stimulation of cutaneous and visceral organs indicates that sensory information reaches the amygdala and orbital cortex. In the past, a large number of studies have thoroughly documented multiple neural pathways by which sensory information can reach these regions via relay nuclei in the brainstem and diencephalon. Recent studies reported that the amygdala and orbital cortex also receive direct input from the spinal cord. The aim of this study was to determine the magnitude and the origin of these projections in the rat. Injections of the retrograde tracer Fluoro-Gold (FG), restricted to the amygdala, labeled several hundred neurons bilaterally (60% contralateral) throughout the length of the spinal cord. More than 60% of labeled neurons were found in the lateral reticulated area of the deep dorsal horn and the gray matter surrounding the central canal. Many neurons were also found in the lateral spinal nucleus. Labeled neurons were concentrated in upper lumbar and upper cervical segments. Injections of Fluoro-Gold that were centered in the orbital cortex labeled only a small number of neurons (73% contralateral) within the spinal cord. Most labeled neurons were found in the lateral reticulated area. Neurons located in the intermediate zone and the gray matter surrounding the central canal were found mainly in upper lumbar and upper cervical segments. These findings, together with the anterograde tracing observations, provide evidence for direct projections of spinal cord neurons to the amygdala and orbital cortex. Their laminar distribution in the spinal cord and the involvement of the amygdala and orbital cortex in limbic functions suggest that these pathways may play a role in neuronal circuits that enable somatosensory information, including pain, to affect autonomic, endocrine and behavioral functions.

Amygdala↗
PubMed1993

Projections from the nucleus tractus solitarii to the spinal cord.

Projections from the nucleus tractus solitarii (NTS) to the spinal cord were demonstrated in the male Sprague-Dawley rat. In retrograde transport studies, a horseradish peroxidase conjugate or a fluorescent dye, FluoroGold, were injected into midcervical or upper thoracic spinal segments. Most solitariospinal neurons were multipolar or bipolar and located between the obex and spinomedullary junction. Solitariospinal neurons were concentrated in proximity to the ventral border of the solitary tract and extended dorsally into the intermediate division and ventrolaterally into the intermediate reticular zone (IRt) of the lateral tegmental field. This subgroup predominantly projects to midcervical spinal segments. A subset of small neurons was retrogradely labeled from cervical or thoracic spinal segments in the medial commissural nucleus and contiguous with a periventricular group surrounding the central canal. In anterograde transport studies, iontophoretic deposits of Phaseolus vulgaris leucoagglutinin were centered stereotaxically on sites in NTS identified by retrograde transport data. The lectin was incorporated by neurons of the solitary complex and transported bilaterally by axons that emerged from the nucleus and entered the reticular formation. The solitario-reticular (transtegmental) pathway irradiated diagonally across the IRt and extended caudally into the cervical lateral funiculus and spinal gray. A small periventricular-spinal pathway also descended longitudinally to the neuraxis. Solitariospinal neurons project to superficial lamina of the dorsal horn, laminae VII and X and ventral horn. The projections are predominantly contralateral to phrenic and intercostal motor nuclei and ipsilateral to the intermediolateral cell column. The solitariospinal projection represents the shortest route in the central nervous system, other than the local intraspinal reflex, through which first order visceral afferents signal cardiorespiratory and alimentary motor nuclei.

Animals↗
PubMed1993

Cerebellar influences on accessory oculomotor nuclei of the rat: a neuroanatomical, immunohistochemical, and electrophysiological study.

With the aim to evaluate a possible neocerebellar control on eye movements, the projections from the cerebellar lateral nucleus (LN) to the accessory oculomotor nuclei (i.e., the nucleus of posterior commissure, the nucleus of Darkschewitsch, and the interstitial nucleus of Cajal), the putative neurotransmitters subserving this pathway, and the nature of the synaptic influences exerted by these projections were studied in adult rats. We used the orthograde transport of horseradish peroxidase conjugated with wheat germ agglutinin (WGA-HRP) to identify the mesencephalic areas where cerebellofugal fibers terminate, and retrograde labeling with the fluorescent dye fluoro-gold to estimate the incidence of cerebellar neurons projecting to the accessory oculomotor nuclei. Orthograde labeling showed that only a small contingent of cerebellofugal fibers reaches the contralateral accessory oculomotor nuclei. The retrogradely labeled cells were located primarily in the small-celled part of LN. By immunohistochemistry, we observed that all the cells retrogradely labeled from the accessory oculomotor area were also stained by using glutamate or aspartate antisera, but none of them were double-stained with a GABA antiserum. Electrical stimulation of the contralateral LN elicited changes in firing rate of a significant fraction of cells belonging to the accessory oculomotor nuclei (36.4% in the nucleus of posterior commissure, 47.1% in the nucleus of Darkschewitsch, and 44.6% in the interstitial nucleus of Cajal). In 57.8% of the cases, the responses were excitations, most of which had latencies and response characteristics compatible with a monosynaptic linkage. The remaining 42.2% of the cases were inhibitions with latencies ranging between 5 and 22 ms. Extracellular field potential recordings within the contralateral accessory oculomotor nuclei were interpreted as arising from impulses propagating along excitatory axons projecting in a bundle from the cerebellum. Stimulation of LN area in rats following intranuclear injection of kainic acid was not capable of evoking short latency excitations, so these responses can be considered to depend on the activation of LN efferents. The LN projection on accessory oculomotor nuclei could be part of the final precise control exerted by the neocerebellum on those brain structures concerned with movements of the eyes.

Animals↗
PubMed1993

The patterns of afferent innervation of the core and shell in the "accumbens" part of the rat ventral striatum: immunohistochemical detection of retrogradely transported fluoro-gold.

Recent data have emphasized the neurochemically distinct nature of subterritories in the accumbens part of the rat ventral striatum termed the core, shell, and rostral pole. In order to gain a more comprehensive understanding of how afferents are distributed relative to these subterritories, immunohistochemical detection of retrogradely transported Fluoro-Gold was carried out following iontophoretic injections intended to involve selectively one of the subterritories. The data revealed that a number of cortical afferents of the medial shell and core originate in separate areas, i.e., the dorsal peduncular, infralimbic, and posterior piriform cortices (to medial shell) and the dorsal prelimbic, anterior agranular insular, anterior cingulate, and perirhinal cortices (to core). The lateral shell and rostral pole are innervated by cortical structures that also project either to the medial shell or core. The orbital, posterior agranular insular, and entorhinal cortices, hippocampus, and basal amygdala were observed to innervate the accumbens in a topographic manner. Following core injections, strong bilateral cortical labeling was observed. Few labeled cortical cells were observed contralaterally following injections in the medial shell. Intermediate numbers of labeled neurons were observed in contralateral cortices following lateral shell injections. Robust subcortical labeling in a variety of structures in the ventral forebrain, lateral hypothalamus, deep temporal lobe, and brainstem was observed after shell injections, particularly those that involved the caudal dorsomedial extremity of the shell, i.e., its "septal pole." Selective ipsilateral labeling of subcortical structures in the basal ganglia circuitry was observed following injections in the core and, to a lesser extent, lateral shell. It was concluded that a number of afferent systems exhibit varying degrees of segregation with respect to the accumbal subterritories.

Afferent Pathways↗
PubMed1993

Efferent projections of the sexually dimorphic area of the gerbil hypothalamus: anterograde identification and retrograde verification in males and females.

Outputs of the sexually dimorphic area (SDA) of the gerbil hypothalamus were identified by injecting Phaseolus vulgaris-leucoagglutinin into the medial or lateral SDA (mSDA, lSDA) in males and females. They were verified by injecting Fluoro-Gold or rhodamine-labeled beads into over half the areas that contained labeled fibers. Both anterograde and retrograde tracing showed that the mSDA and lSDA project to many of the same sites but often to differing degrees. The mSDA projects more heavily than the lSDA to many of their forebrain targets including the ventral part of the lateral septal nucleus, the bed nucleus of the stria terminalis, the medial tuberal area, and the anteroventral periventricular, arcuate, ventromedial and ventral premammillary nuclei of the hypothalamus. The lSDA projects more heavily than the mSDA to many of their mid- and hindbrain targets including the caudal, ventrolateral part of the periaqueductal gray, the retrorubral field, the pedunculopontine tegmental nucleus, and the locus coeruleus. In many other areas of the brain, the projections of the mSDA and lSDA are similar in size. These areas include the substantia innominata, the vascular organ of the lamina terminalis, the anterior amygdala, the posterior hypothalamus, the reuniens and paraventricular nuclei of the thalamus, and the pontine periaqueductal gray lateral to the fourth ventricle. The SDA pars compacta (SDApc), a small cell group embedded in the mSDA of males, projects to many fewer areas than the surrounding mSDA. It was strongly labeled when retrograde tracers were injected into the encapsulated part of the bed nucleus of the stria terminalis, the anteroventral periventricular nucleus, or the mSDA. It was also labeled from the vascular organ of the lamina terminalis, the caudal part of the lateral bed nucleus of the stria terminalis, the lSDA, the area lateral to the mSDA, the arcuate nucleus, the ventral premammillary nucleus, and the ventrolateral part of the ventromedial nucleus of the hypothalamus. Nothing resembling an SDApc was identified during retrograde tracing in females.

Animals↗
PubMed1994

Comparative efficacy of expression of genes delivered to mouse sensory neurons with herpes virus vectors.

To achieve gene delivery to sensory neurons of the trigeminal ganglion, thymidine kinase-negative (TK-) herpes simplex viruses (HSV) containing the reporter gene lacZ (the gene for E. coli beta-galactosidase) downstream of viral (in vectors RH116 and tkLTRZ1) or mammalian (in vector NSE-lacZ-tk) promoters were inoculated onto mouse cornea and snout. Trigeminal ganglia were removed 4, 14, 30, and 60 days after inoculation with vectors and histochemically processed with 5-bromo-4-chloro-3 indolyl-beta-galactoside (X-Gal). With vector tkLTRZ1, large numbers of labeled neurons were observed in rostromedial and central trigeminal ganglion at 4 days after inoculation. A gradual decline in the number of labeled neurons was observed with this vector at subsequent time points. With vectors RH116 and NSE-lacZ-tk, smaller numbers of labeled neurons were seen at 4 days following inoculation than were observed with vector tkLTRZ1. No labeled neurons could be observed at 14 days after inoculation with vectors RH116 and NSE-lacZ-tk. Immunocytochemistry for E. coli beta-galactosidase and in situ hybridization to HSV latency-associated transcripts revealed labeled neurons in regions of the trigeminal ganglion similar to that observed with X-Gal staining. A comparable distribution of labeled neurons in trigeminal ganglion was also observed after application of the retrograde tracer Fluoro-Gold to mouse cornea and snout. These data provide evidence that retrogradely transported tk- herpes virus vectors can be used to deliver a functional gene to sensory neurons in vivo in an anatomically predictable fashion.

Animals↗
PubMed1994

Grafts of fetal central nervous system tissue rescue axotomized Clarke's nucleus neurons in adult and neonatal operates.

Many conditions are thought to contribute to neuron death after axotomy, including immaturity of the cell at the time of injury, inability to reestablish or maintain target contact, and dependence on trophic factors produced by targets. Exogenous application of neurotrophic factors and transplants of peripheral nerve and embryonic central nervous system (CNS) tissue temporarily rescue axotomized CNS neurons, but permanent rescue may require transplants that are normal targets of the injured neurons. We examined the requirements for survival of axotomized Clarke's nucleus (CN) neurons. Two months after hemisection of the spinal cord at the T8 segment, there was an ipsilateral 30% loss of neurons at the L1 segment in adult operates and a 40% loss in neonates. Transplants of embryonic spinal cord, cerebellum, and neocortex inserted into the T8 segment at the time of hemisection prevented virtually all of the cell death in both adults and neonates, but transplants of embryonic striatum were ineffective. None of the grafts prevented the somal atrophy of CN neurons caused by axotomy. Retrograde transport of fluoro-gold from the cerebellum demonstrated that 33% of all CN neurons at L1 project to the cerebellum, 50% of these died following a T8 hemisection, but all these projection neurons were rescued by a transplant of embryonic spinal cord. These results suggest that the rescue of axotomized CN neurons is relatively specific for the normal target areas of these neurons, but this specificity is not absolute and may depend on the distribution and synthesis of particular neurotrophic agents.

Aging↗
PubMed1994

Ventral root avulsion: an experimental model of death of adult motor neurons.

The present study proposes a reproducible model of experimental degeneration of adult motor neurons in the rat. Avulsion of ventral roots in the adult lumbar cord transects motor axons at the root exit and leads to retrograde cell death of 80% of motor neurons 2 weeks later; this result follows a series of retrograde changes, including chromatolysis, loss of transmitter phenotype, and accumulation of phosphorylated neurofilaments in perikarya. Glial cells recruited at the site of retrograde injury express both microglia-specific epitopes (as exemplified by OX-42 immunoreactivity) and macrophage-specific markers (e.g., ED-1 immunoreactivity). Macrophage-specific markers become particularly intense 7 days postaxotomy and provide additional evidence of active phagocytosis of injured neurons. Ventral root avulsion is a very useful model for assessing mechanisms of motor neuron death and testing the ability of trophic factors and other agents to preserve the phenotype and promote the survival of adult motor neurons in vivo.

Animals↗