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PubMed1991

Substance P-immunoreactive innervation from the retina to the suprachiasmatic nucleus in the rat.

The present study examined substance P (SP) innervation in the suprachiasmatic nucleus (SCN) of the rat. In the colchicine-untreated rat, SP-immunoreactive fibers formed a dense oval plexus in the ventral part of the SCN. After bilateral eye enucleation, there was a marked reduction in SP-immunoreactive fibers in the ventral part of the SCNs. The SP-immunoreactive neurons in the retinal ganglion cell layer were retrogradely labeled after injection of Fluoro-gold into the SCN. These findings indicate the presence of the SP innervation from the retina to the SCN in the rat. The role of SP in the retino-hypothalamic tract was discussed from the light-dark cycle.

Animals↗
PubMed1991

Projections of the paraventricular nucleus of the hypothalamus to the sexually dimorphic lumbosacral region of the spinal cord.

Lumbar regions L5-L6 of the spinal cord of the male rat contain the sexually dimorphic motor nuclei, the spinal nucleus of the bulbocavernosus (SNB) and the dorsolateral nucleus (DLN), which innervate perineal muscles, the bulbocavernosus and the ischiocavernosus, respectively. This neuromuscular system controls penile reflexes which are essential to male reproductive success. Oxytocin has been shown to induce penile reflexes and the site of action for these effects is the PVN. Since PVN is known to project to cervical and thoracic levels of spinal cord, the present study examined projections of the PVN to the L5-L6 region of the spinal cord. WGA-HRP was injected into the region of L5-L6, aimed at the SNB and DLN and their dendritic extents, in intact male, castrated male and female rats. WGA-HRP-labelled cells bodies were found in the parvocellular subnuclei of PVN, as well as regions of the lateral hypothalamus and the dorsal area of the hypothalamus. These results demonstrate that the PVN projects to lumbar levels of the spinal cord that are sexually dimorphic and androgen-dependent. This suggests that PVN may modulate the activity of these motoneurons.

Animals↗
PubMed1992

CRF-like immunoreactivity selectively labels preganglionic sudomotor neurons in cat.

Immunohistochemical and neuronal tracing methods were used in cats to determine which type of postganglionic sympathetic neuron is innervated by preganglionic neurons which contain corticotrophin releasing factor-like immunoreactivity (CRF-LI). Preganglionic neurons with CRF-LI have their cell bodies at two restricted levels of the spinal cord and terminate in the stellate and lower lumbar ganglia. CRF-LI terminal baskets in stellate and lumbar ganglia surrounded cell bodies, 96-99% of which showed no tyrosine hydroxylase (TH)-LI (presumptive cholinergic neurons). Calcitonin gene-related peptide (CGRP)-LI was used to label the cholinergic ganglion cells which innervate sweat glands: 96-99% of those were confirmed as lacking TH-LI, while the remainder showed weak staining. Every one of over 6000 CRF-LI terminal baskets counted in 4 stellate and 6 lumbar ganglia was found to surround a cell body with CGRP-LI; conversely, 81-86% of the cell bodies showing CGRP-LI were surrounded by CRF-LI terminal baskets. In 3 cats, the retrograde tracer fluorogold was used to label postganglionic neurons projecting to the paw pads (a population which includes both cholinergic sudomotor neurons and noradrenergic vasoconstrictor neurons). Between 26 and 38% of the retrogradely labelled ganglion cells were surrounded by CRF-LI terminal baskets. We conclude that in cats, preganglionic sympathetic neurons with CRF-LI are sudomotor in function.

Acetylcholine↗
PubMed1992

Differential sites of origin and collateralization of corticospinal neurons in the rat: a multiple fluorescent retrograde tracer study.

Cells of origin for corticospinal fibers in the rat were identified following retrograde transport of Fluoro-Gold (FG), Propidium iodide (PI), Fast blue (FB), and Diamidino yellow (DY) injected unilaterally into lumbar (FG), mid-thoracic (PI), cervical enlargement (FB), and cranial cervical (DY) spinal gray matter. Most labeled neurons were contralateral to injection in lamina V and ranged from small to very large. These cells occupied two distinct cortical regions: one rostral and the other larger and more caudal. Neurons of the rostral region projected axons solely to cervical spinal segments whereas neurons of the caudal region projected fibers to all spinal segments. Somatotopically, most neurons projecting to lumbar segments were most medial. More than 98% of all labeled cortical neurons contained only a single fluorescent tracer; however, within a single tissue section each of the 4 tracers could be found in these single labeled neurons. The few double labeled neurons contained only cervical (DY + FB) or thoracolumbar (PI + FG) tracers. No triple or quadruple labeled cells were seen. Hence morphological evidence is presented that corticospinal axons branch to terminate in more than one spinal region, but these collateral terminations are restricted to only a few adjacent spinal segments.

Amidines↗
PubMed1992

Basic fibroblast growth factor-like immunoreactivity in the trigeminal proprioceptive and motor systems.

Basic fibroblast growth factor (bFGF) isolated from the brain and pituitary, has been shown to induce cell divisions in a variety of cell types. It also acts as a potent stimulator of angiogenesis, and it is important in the survival of several types of cultured neurons. Despite considerable information on the functions of bFGF, there is incomplete knowledge about the ways in which it reaches remote tissues and its subcellular localization in the adult brain. Here we report our findings that a certain population of neurons with free ribosomes and rough endoplasmic reticulum immunoreactive for bFGF in the mesencephalic nucleus of the trigeminal nerve sends proprioceptive fibers to muscle spindles in the masseter muscle, and immunoreactive axons to the trigeminal motor nucleus to form synapses with the bFGF-containing motoneurons whose axons further constitute myoneural junctions in the periphery. Moreover, some bFGF neurons contain electron dense immunoreaction deposits in the euchromatin but not in the heterochromatin of the nucleus. These findings suggest that endogenous bFGF is transported within nerve processes and functions in mature neuronal circuits subserving the masseteric reflex arcs, and that bFGF is produced in free ribosomes and/or rough endoplasmic reticulum and is transported into the genetically active euchromatin as well.

Animals↗
PubMed1992

Cholecystokinin induces c-fos expression in hypothalamic oxytocinergic neurons projecting to the dorsal vagal complex.

Systemic administration of cholecystokinin (CCK) decreases gastric motility and stimulates pituitary secretion of oxytocin (OT). Although peripheral OT does not affect gastric function, increasing evidence suggests that central OT secretion acting within the dorsal vagal complex (DVC) can alter gastric motility. To evaluate whether systemically administered CCK is capable of activating oxytocinergic neurons projecting to the DVC, we utilized fluorogold retrograde labeling from the DVC in combination with c-fos and OT immunocytochemical staining to quantitatively analyze paraventricular nucleus (PVN) neurons of rats following injection of CCK at a dose known to cause maximal pituitary OT secretion (100 micrograms/kg i.p.). Our results showed that 2320 +/- 63 PVN neurons were retrogradely labeled from the DVC; 146 +/- 21 (6.3%) of these contained OT, and these cells were predominantly located in the medial parvocellular subdivision of the PVN. Of all retrogradely labeled cells, 671 +/- 112 (28.9%) expressed c-fos after CCK stimulation, and 68 +/- 14 of these (10.1%) contained OT. Approximately 50% of the OT-containing neurons retrogradely labeled from the DVC stained positively for c-fos. Many magnocellular OT neurons in the PVN that were not retrogradely labeled from the DVC also expressed c-fos after CCK stimulation. These results demonstrate that parvocellular OT neurons projecting to the DVC are co-activated along with magnocellular OT neurons projecting to the pituitary following administration of a large dose of CCK, and lend support to a possible functional role for OT as a central neurotransmitter that modulates vagal efferent traffic to the gastrointestinal tract.

Animals↗
PubMed1992

Studies related to the use of colchicine as a neurotoxin in the septohippocampal cholinergic system.

Colchicine has been shown to be neurotoxic to cholinergic neurons in the medial septum 1 week following intracerebroventricular injections. The experiments described here were designed to examine the selectivity of this effect over a longer time course, and to examine the role of axoplasmic transport in the neurotoxic effect. As previously reported, 1 week after intracerebroventricular injections of colchicine, the numbers of choline acetyltransferase (ChAT)-immunoreactive neurons in the medial septum-diagonal band complex (MSDB) were reduced to 38% of control; this reduction was stable 2 and 3 weeks post injection. Injections of colchicine placed into the body of the fornix produced similar results. GAD-immunoreactive somata, the other major population of neurons in the MSDB, were unaffected 3 weeks following colchicine, as previously reported 1 week following similar injections. The normal AChE staining pattern in the hippocampus, particularly the dentate gyrus, was depleted following either ICV or intrafornical injections of colchicine. This depletion was more severe with longer survival times. Injections of lumicolchicine, an isomer of colchicine which does not bind tubulin, had no effect on ChAT-immunoreactive neurons in the MSDB or on AChE staining in the hippocampus. Injections of colchicine, but not of lumicolchicine, partially blocked the retrograde transport of the fluorescent dye Fluoro-Gold from the hippocampus to the MSDB. In addition, the content of NGF in the hippocampus rose 84% above control values 2 weeks following colchicine and remained elevated at three weeks. Together these results indicate that colchicine is selectively toxic for cholinergic neurons in the septohippocampal system, and suggest that the alkaloid's neurotoxic effects work via the blockade of axoplasmic transport.

Acetylcholine↗
PubMed1992

Neurons in the median preoptic nucleus of the rat with collateral branches to the subfornical organ and supraoptic nucleus.

A novel pathway between the subfornical organ and the supraoptic nucleus involving the collateral branches of cell bodies situated in the lamina terminalis has been studied. Fluorogold was injected into the supraoptic nucleus and rhodamine-labelled microspheres into the subfornical organ of rats. Nineteen % of neurons in the median preoptic nucleus and 30% of neurons in the OVLT projecting to the subfornical organ also had axons extending to the supraoptic nucleus. These pathways may represent a novel trajectory for the rely of information from the lamina terminalis to the supraoptic nucleus.

Animals↗
PubMed1993

Glutaminase-like immunoreactivity in rat spinomesencephalic tract cells.

Retrograde transport of the fluorescent tracer Fluorogold was used in combination with immunohistochemical staining for the enzyme glutaminase to identify putative glutamatergic neurons belonging to the rat spinomesencephalic tract. Glutaminase-like staining in spinal projection neurons suggests that the relay of nociceptive information from the spinal cord to midbrain may involve the excitatory amino acid glutamate.

Animals↗
PubMed1993

Origin of noradrenergic projections to GnRH perikarya-containing areas in the medial septum-diagonal band and preoptic area.

The purpose of the present study was to identify the sites of origin of the noradrenergic fibers that project to areas containing gonadotropin-releasing hormone (GnRH) perikarya since norepinephrine (NE) is known to influence the activity of GnRH neurons. Fluorescent retrograde tracers were used in combination with immunohistochemistry for dopamine-beta-hydroxylase (DBH) and GnRH. Small volumes of either Fluoro-gold (FG) or Fluoro-Ruby (FR) were pressure injected into areas that contain the largest number of GnRH cell bodies, i.e., the medical septum-diagonal band complex or preoptic area. Retrogradely labeled neurons were observed ipsilaterally in the following noradrenergic cell groups: A2 (in the nucleus tractus solitarii), A1 (in the ventrolateral medulla) and locus coeruleus. Approximately 8% of all DBH-positive neurons within the A2-cell group were retrogradely labeled, while 12% of DBH-ir neurons in the A1-group were double-labeled. Only a few retrogradely labeled DBH-ir neurons were observed in the locus coeruleus (< 1%). Double-labeled neurons were not organized into discrete cell groups, but were dispersed among other NE-neurons within the A2- and A1-cell groups. The highest concentrations of double-labeled neurons were located in the central one-third of both the A2 and A1 cell groups. The results suggest that most noradrenergic terminals in the region of the GnRH perikarya in the medial septum-diagonal band/rostral preoptic area originate from ipsilateral neurons in areas A1 and A2.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

The retrograde tracer Fluoro-Gold interferes with the infectivity of herpes simplex virus.

Fluoro-Gold has been used previously to identify those trigeminal ganglion cells that innervate the central cornea. To examine the effects of Fluoro-Gold treatment on infection and spread of HSV in vivo, we measured the number of plaque forming units recovered from trigeminal ganglia 3 or 5 days after corneal scratch and inoculation with Fluoro-Gold and HSV. Treatment with Fluoro-Gold reduced the amount of virus recovered after retrograde transport 63% at 3 days and 28% at 5 days after inoculation. When we examined trigeminal ganglion sections from animals treated with HSV and Fluoro-Gold, we found the number of neurons double labeled with antibodies that recognize HSV and Fluoro-Gold was only 13% of all Fluoro-Gold labeled neurons. This was significantly fewer cells that we had anticipated, on the basis of double labeling experiments with wheat germ agglutinin combined with Fluoro-Gold. The effects of varying doses of the retrograde tracer, Fluoro-Gold on Herpes simplex virus (type 1) (HSV) infectivity were also assayed in vitro using a standard viral plaque assay. At 1 x 10(-3) mg/ml Fluoro-Gold there was no effect on the number of plaque forming units. At 5 x 10(-1) mg/ml the number of plaques was reduced about 67%. We conclude that Fluoro-Gold interferes with productive HSV infection in vivo and in vitro after retrograde transport of HSV by neurons.

Animals↗
PubMed1993

The projections of noradrenergic neurons in the A5 catecholamine cell group to the spinal cord in the rat: anatomical evidence that A5 neurons modulate nociception.

Brainstem noradrenergic neurons located in the A5, A6, and A7 catecholamine cell groups provide the entire noradrenergic innervation of the spinal cord. We have previously demonstrated that noradrenergic neurons in the A6 and A7 cell groups innervate the ventral and dorsal horns, respectively. Since the specific spinal cord terminations of the A5 cell group have not been clearly delineated, the present experiments were designed to trace the projections from this noradrenergic cell group to the spinal cord, using the anterograde tracer Phaseolus vulgaris-leucoagglutinin (PHA-L) in combination with dopamine-beta-hydroxylase immunocytochemistry. The results of these experiments indicate that A5 noradrenergic neurons project ipsilaterally through the dorsolateral funiculus in cervical, thoracic, and lumbar segments. In cervical segments, these axons terminate primarily in the ipsilateral deep dorsal horn (laminae IV-VI) and the intermediate zone (lamina VII). In thoracic segments, the intermediolateral cell column is heavily innervated by A5 axons. In lumbar segments, the concentration of A5 axons is more diffuse and more widely distributed than that in cervical and thoracic segments. Although there is a higher density of axons in the deep dorsal horn and the intermediate zone, there are also scattered axons in the dorsal and ventral horns. The innervation of these regions of the spinal cord by A5 neurons provides anatomical support for the conclusion that these noradrenergic neurons are involved in modulating cardiovascular reflexes and nociceptive transmission in the spinal cord.

Animals↗
PubMed1993

Peripheral and central pathways regulating the kidney: a study using pseudorabies virus.

We used the retrograde transneuronal transport of a neurotropic virus, pseudorabies virus (PRV), to identify the neurons in sympathetic ganglia, spinal cord and brain which regulate renal function and renal circulation. PRV was microinjected into the left kidney of 70, pentobarbital-anesthetized, male rats. After an incubation period of 1-4 days, rats were anesthetized and sacrificed. PRV-infected neurons were located immunocytochemically in pre- and paravertebral sympathetic ganglia, the intermediolateral cell column of the T10-T13 segments and several brainstem cell groups: the medullary raphe nuclei, rostral ventrolateral medulla, rostral ventromedial medulla, A5 cell group, and the paraventricular hypothalamic nucleus. In more heavily infected rats, additional labeling was found in the locus coeruleus, periaqueductal gray matter, lateral hypothalamic area, zona incerta, and anterior hypothalamic area. No infected propriospinal neurons were observed in the lateral spinal nucleus or gray matter of the caudal cervical, lumbosacral or thoracic spinal segments not containing infected putative sympathetic preganglionic neurons. The paucity of infected propriospinal neurons in the presence of infected brainstem neurons, even in lightly infected rats, is discussed in reference to the relative importance of descending vs spinal regulation of the sympathetic outflow to the kidney.

Animals↗
PubMed1993

Neurogenetical segregation of the vestibulospinal neurons in the rat.

The time of origin of the vestibulospinal projection neurons was determined by a double-labeling method using 5-bromodeoxyuridine (BrdU), the thymidine analogue, and Fluoro-Gold (FG), a retrograde fluorescent tracer. Rat fetuses were exposed to BrdU in utero to label the vestibular neurons on one of the embryonic (E) days between E12 and E15. Upon reaching adulthood, the rats were given unilateral injections of FG into the cervical cord to identify the spinal projection neurons. Brainstem sections were immunohistochemically processed for BrdU and then examined for neurons that were both BrdU-positive and FG-positive in the vestibular nuclei. In the lateral vestibular nucleus (LVe), most of the vestibulospinal neurons were generated on E12. In the inferior vestibular nucleus (IVe), the vestibulospinal neurons were produced almost equally on both E12 and E13. In the medial vestibular nucleus (MVe), the vestibulospinal neurons were generated consistently on days between E12 and E14 with a mild peak on E13. The present results thus demonstrate that genesis of the vestibulospinal neurons occurs sequentially in the following order: firstly in the LVe, secondly in the IVe, and finally in the MVe. The different sequential generation of vestibulospinal neurons among the LVe, MVe and IVe may reflect the fact that the vestibulospinal projections are differentially organized depending on the nature of each subnucleus.

Animals↗
PubMed1993

Differences in Fluorogold and wheat germ agglutinin-horseradish peroxidase labelling of bladder afferent neurons.

Rat urinary bladder afferent neurons were significantly smaller (34%) when labelled with Fluorogold (FG) than when labelled with wheat germ agglutinin-horseradish peroxidase (WGA-HRP). This study showed that this difference was due to an artifact of tissue processing (ethanol dehydration) and was not due to uptake and transport of the two tracers by two different subpopulations of bladder afferents.

Afferent Pathways↗
PubMed1993

The subdivisions of the intermediolateral nucleus in the sacral spinal cord of the cat.

The subdivisions of the sacral intermediolateral nucleus (IML) of the cat have been studied by using a double-labeling technique of retrograde Fluoro-gold (FG) and wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) tracing. The parasympathetic preganglionic neurons (PGNs) that were labeled by the FG injected into the pelvic nerve formed a 'V'-shaped column known as the sacral parasympathetic nucleus (SPN) in the sacral IML. The neurons that were labeled by the WGA-HRP applied to the lateral parabrachial nucleus (PBL) formed an elongated spindle-shaped column extending throughout the IML of the sacral segments. We designated it by the name of sacral visceral sensory nucleus (SVSN). These findings indicate that the sacral IML of the cat contain two distinct subdivisions, SPN and SVSN.

Animals↗
PubMed1993

Localization of NADPH diaphorase in neurons of the rostral ventral medulla: possible role of nitric oxide in central autonomic regulation and oxygen chemoreception.

We studied whether neurons containing nitric oxide synthase (NOS) are localized to the rostral ventrolateral medulla (RVM) and, if so, whether they are distinct from the adrenergic neurons of the C1 group. NOS-containing neurons and/or C1 neurons were visualized using NADPH diaphorase histochemistry and phenylethanolamine N-methyltransferase (PNMT) immunohistochemistry, respectively. A column of NADPH diaphorase-positive neurons, extending 2 mm in the rostrocaudal plane, was observed lateral to the inferior olive and medial to the C1 neurons. Double labelling studies showed that NADPH diaphorase-positive neurons were not immunoreactive for PNMT, indicating that the two enzymes were localized in the different cells. Furthermore, only a small fraction of NADPH diaphorase neurons were retrogradely labelled after injections of fluorogold into the thoracic cord. We conclude that the RVM contains a well-defined group of neurons endowed with NOS that are distinct from the adrenergic neurons of the C1 group and have only limited monosynaptic projections to the spinal cord. Since the RVM is involved in the control of arterial pressure and in oxygen-conserving reflexes, the findings raise the possibility that nitric oxide participates in central autonomic regulation and oxygen chemoreception.

Animals↗
PubMed1993

Calretinin-immunoreactivity in trigeminal neurons innervating the nasal mucosa of the rat.

Trigeminal primary neuronal cell bodies were labeled by retrograde transport of Fluoro-gold (FG) from the nasal mucosa of rats. The trigeminal ganglion containing the labeled cell bodies were processed for double stain for calretinin- and tachykinin-immunoreactivities (CR- and TK-irs). Except for a few contralateral cells, all the cells that innervated the nasal mucosa (NM cells) were confined to the ophthalmo-maxillary division of the trigeminal ganglion ipsilateral to the FG application. In the dorsal two-thirds of the ganglion, NM cells formed a cluster in the rostromedial part of ophthalmo-maxillary division (the rostromedial cluster). In the ventral third, the number of cells in the rostromedial cluster markedly decreased. Instead, numerous NM cells were found in the caudolateral part of the ophthalmo-maxillary division (the caudoventrolateral cluster). CR- and TK-irs were detected in 18% and 54% of overall population of NM cells, respectively. Virtually all of CR-immunoreactive (-ir) NM cells coexpressed TK. Although the proportion of TK-ir cells, irrespective of CR-ir, was similar for both clusters, CR-ir cells were more frequent in the caudoventrolateral cluster than in the rostromedial cluster. In the dorsal 1/3 of the ganglion where all the NM cells belonged to the rostromedial cluster, only 8.4% exhibited CR-ir. On the other hand, as much as 30.1% of NM cells expressed CR-ir in the ventral 1/3 where most NM cells were found in the caudoventrolateral cluster. Trigeminal cell bodies innervating the cornea and conjunctivum were located in the rostromedial part of the ophthalmo-maxillary division.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗