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PubMed1996

Effects of neonatal exposure to anti-nerve growth factor on the number and size distribution of trigeminal neurones projecting to the molar dental pulp in rats.

The first aim of the present study was to determine whether depletion of endogenous nerve growth factor (NGF) during early postnatal development results in a long-term deficit in the number of trigeminal ganglion cells and axons projecting to the molar pulp. The second aim was to identify selectivity of the effects of NGF deprivation for any specific size group among pulp neurones. Newborn Sprague-Dawley rats were given subcutaneous injections of either rabbit anti-mouse-NGF serum or non-immune (control) rabbit serum for a period of 1 month. At age 4 months, Fluoro-gold (FG) was applied to the pulp chamber of the right maxillary first molar. One week later the animals were perfusion-fixed, and the trigeminal ganglia were removed and serially sectioned with a cryostat. Labelled neurones were seen only in the trigeminal ganglia ipsilateral to the injected teeth. The area of every labelled cell profile was measured, and from these data, estimates of the true number and size distribution of FG-labelled cells were obtained by recursive translation. Ganglia of control animals had a mean of 197 labelled neurones, all in the maxillary division, and most of the somas were of medium or large diameter. NGF-deprived animals had significantly fewer (mean = 145) FG-labelled cells in the trigeminal ganglion ipsilateral to the injected tooth. Neurones with somas of less than 30 microns dia were most strikingly subnormal in anti-NGF treated animals (64% of controls). In accordance with the greater susceptibility of small neurones to anti-NGF exposure, deficits in apical nerve fibres of the mandibular first molar were greater in degree and duration for unmyelinated axons than for myelinated axons. It is concluded that NGF is an important mediator in regulation of postnatal development of the sensory innervation of the dental pulp. The results also indicate that postnatal development of at least one class of larger pulpal afferent neurones is regulated by factors other than NGF.

Animals↗
PubMed1986

Fluoro-Gold: a new fluorescent retrograde axonal tracer with numerous unique properties.

A new fluorescent dye, Fluoro-Gold, has been demonstrated to undergo retrograde axonal transport. Its properties include intense fluorescence, extensive filling of dendrites, high resistance to fading, no uptake by intact undamaged fibers of passage, no diffusion from labeled cells, consistent and pure commercial source, wide latitude of survival times and compatibility with all other tested neuro-histochemical techniques.

4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfo↗
PubMed1987

Neurons in the area postrema are the only catecholamine-synthesizing cells in the medulla or pons with projections to the rostral ventrolateral medulla (C1-area) in the rabbit.

We have identified, in the rabbit medulla and pons, neurons which project to the C1-region of the rostral ventrolateral medulla. By combining tyrosine hydroxylase immunohistochemistry with retrograde transport of Fluoro-Gold we determined whether any of the retrogradely labelled neurons synthesize catecholamines. The only doubly labelled cells were located in the area postrema. No other group of catecholamine-synthesizing neurons in either the medulla or the pons was found to project to the C1-area of the rostral ventrolateral medulla. Pharmacological agents which lower arterial pressure by stimulating adrenoceptors in the rostral ventrolateral medulla may act on receptors which are not innervated by catecholamine-synthesizing perikarya located outside the C1-region.

Animals↗
PubMed1988

Dopaminergic nigrotectal projection in the rat.

After injecting a fluorescent tracer (Fluoro-gold) into the rat superior colliculus, retrogradely labeled neurons in the rostral, ventrolateral portions of the substantia nigra pars reticulata were also immunohistochemically labeled with tyrosine hydroxylase antisera. 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) pretreatment of the medial forebrain bundle resulted in the disappearance of the nigral neurons double-labeled with the two markers. The existence of a dopaminergic nigrotectal projection susceptible to MPTP might provide a morphological substrate for abnormal saccadic eye movements in parkinsonism.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗
PubMed1988

Labeling of nerve cells in the dorsal motor nucleus of the vagus of rats by retrograde transport of Fluoro-Gold.

Nerve cells in the dorsal motor nucleus of the vagus (dmnX) were identified by retrograde axonal transport after injections of a fluorescent tracer, Fluoro-Gold, into the anterior gastric wall. The intramural injection resulted in labeling of cells in the medial half of the left dmnX. These observations were contrasted with the diffuse (mediolateral and rostrocaudal) and bilateral distribution of labeled cells after Fluoro-Gold solution was dripped onto the stomach. In comparison with other neurotracers, the advantages of Fluoro-Gold are that (1) it can be visualized without the chemical reaction with chromogen, thereby allowing better reproducibility, and (2) it does not fade up to one year.

Animals↗
PubMed1988

Spinal origin of sympathetic preganglionic neurons in the rat.

The segmental distribution of sympathetic preganglionic neurons (SPNs) and dorsal root ganglion cells (DRGs) was studied after Fluoro-gold injections into the major sympathetic ganglia and adrenal gland in rats. A quantitative assessment of the segmental and nuclear locations was made. Four general patterns of innervation were apparent: (1) a large number of SPNs (1000-2000/ganglion) innervate the sympathetic ganglia which control head or thoracic organs and a relatively small number of SPNs (100-400/ganglion) innervate the sympathetic ganglia controlling the gut, kidney, and pelvic organs; this difference in density of innervation probably relates to the level of fine control that can occur in these end organs by the SPNs; (2) the reverse pattern is seen in the DRG labeling where a large number of DRGs were labeled after Fluoro-gold injections into the preaortic ganglia (celiac, superior, and inferior mesenteric) and a small number were labeled after injections into the cervical sympathetic ganglia; (3) the intermediolateral cell column is the main source of SPNs except for the inferior mesenteric ganglion which is innervated predominantly by SPNs originating in the central autonomic nucleus (75%); the lateral funiculus is a source of SPNs mainly for the cervical sympathetic ganglia; and (4) each sympathetic ganglion and the adrenal gland receives a multisegmental SPN and DRG input with one segment being the predominant source of the innervation. The adrenal gland shows an intermediate position in terms of the density of SPN input (approximately 800 cells) and dorsal root input (approximately 300 cells); it has a widespread segmental input (T4-T12) with the T8 segment being the major source.

Animals↗
PubMed1988

The iontophoretic application of Fluoro-Gold for the study of afferents to deep brain nuclei.

A method is described for identifying the afferents to confined areas within the central nervous system using iontophoretic application of the fluorescent tracer, Fluoro-Gold (FG). Unlike other fluorescent tracers, it is possible to make focal iontophoretic injections through small-tipped micropipettes, and electrophysiological recordings from the injection pipette can be used to define structures prior to injections. Retrograde labeling with FG appears to be as sensitive as wheatgerm agglutinin-conjugated horseradish peroxidase visualized with tetramethylbenzidine. Furthermore, iontophoretically applied FG does not appear to be taken up and transported retrogradely by fibers of passage. Finally, retrograde transport of FG can be combined with immunofluorescence without appreciable loss of sensitivity in either label.

Animals↗
PubMed1988

Distinct adenosine deaminase-containing inputs to the substantia nigra from the striatum and tuberomammillary nucleus.

Immunohistochemical, neuroanatomical and lesion methods were used to investigate the projections of adenosine deaminase immunoreactive (ADA-IR) neurons in the striatum (caudate/putamen) and hypothalamus to the substantia nigra (SN). Striatal ADA-IR neurons were distributed within two zones; anteriorly in the medial and ventromedial extreme of the head and body of the striatum, and posteriorly in the tail of the striatum. The posterior hypothalamus contained ADA-positive neurons which were confined to the tuberomammillary nucleus (TM). The SN was devoid of ADA-positive neurons, but contained two distinct types of ADA-IR fiber terminations. One type was confined to bands located at the ventrolateral and dorsomedial borders of the pars reticulata and consisted of fine puncta. The other type was distributed throughout the SN and consisted of long, beaded fibers. Injections of the retrograde tracer Fluoro-gold (FG) into the SN gave rise to FG-labelling of significant numbers of ADA-IR neurons in both the striatum and TM. Medial SN injections preferentially labelled ADA-IR neurons in the anterior striatum and lateral SN injections labelled posterior ADA-IR striatal neurons. Kainic acid lesions of the anterior medial striatum selectively abolished the punctate ADA-IR band in the medial SN and left the long, ADA-IR nigral fibers in an apparently hypertrophied state. Despite depletion of ADA-IR neurons in the striatum by kainic acid, ADA activity increased significantly at striatal lesion sites. The results suggest that the SN receives two topographically segregated fine terminal fields from striatal ADA-IR neurons, and a substantial innervation from ADA-IR neurons in the TM as well. These findings add to the heterogeneous chemical composition of nigral afferents and are discussed in the context of adenosine neuromodulatory mechanisms in the striatonigral system.

Adenosine Deaminase↗
PubMed1989

Fluoro-Gold injected either subcutaneously or intravascularly results in extensive retrograde labelling of CNS neurones having axons terminating outside the blood-brain barrier.

Subcutaneous or intravascular injections of the dye Fluoro-Gold resulted in widespread labelling of neurones with a distribution and appearance consistent with their being motoneurones, preganglionic autonomic neurones and hypothalamic neurones projecting to the pituitary. Peripheral nerve ligation prevented labelling of neurones whose axons travelled in that peripheral nerve. These observations suggest that Fluoro-Gold diffuses extensively in the body tissues but does not cross the blood-brain barrier and that it is taken up by peripheral terminals and transported retrogradely by motoneurones.

Animals↗
PubMed1989

Retrograde labeling of neurons in spinal cord that project directly to nucleus accumbens or the septal nuclei in the rat.

Somatosensory information has been thought to ascend from the spinal cord to limbic areas of the telencephalon through indirect, multisynaptic pathways. We now report that injections of Fluoro-gold into either the nucleus accumbens or the septal nuclei labeled hundreds of neurons in the spinal cord. These and our recent anterograde tracing experiments indicate that some spinal cord neurons project directly to the telencephalon and suggest that nucleus accumbens and septal nuclei process somatosensory information.

Afferent Pathways↗
PubMed1989

Dopaminergic neurons in the nucleus raphe dorsalis innervate the prefrontal cortex in the rat: a combined retrograde tracing and immunohistochemical study using anti-dopamine serum.

The existence of a dopaminergic projection from the nucleus raphe dorsalis (RD) to the prefrontal cortex of the rat was demonstrated by combining immunohistochemistry with retrograde tracing. Injection of horseradish peroxidase or Fluoro-gold into the medial prefrontal cortex resulted in the appearance of retrogradely labelled neurons in the RD and in other brain regions. Some of these retrogradely labelled RD neurons were shown to be immunoreactive for dopamine. These data support the view that these dopaminergic RD neurons represent an extension of the A10 cell group.

Animals↗
PubMed1989

Selectivity of the hippocampal projection to the prelimbic area of the prefrontal cortex in the rat.

Afferent connections of the medial and lateral prefrontal cortex of the rat arising from the hippocampal formation were investigated using iontophoretic application of the fluorescent tracer, Fluoro-gold. Our results demonstrate that the projection which originates in the temporal part of the CA1 hippocampal field and in the prosubiculum is restricted to the prelimbic area of the prefrontal cortex.

Animals↗
PubMed1989

Dynorphin A(1-8) immunoreactive cell bodies, dendrites and terminals are postsynaptic to calcitonin gene-related peptide primary afferent terminals in the monkey dorsal horn.

In the present study, Dynorphin A(1-8) (DYN)-immunoreactive cell bodies, dendrites and terminals are observed postsynaptic to calcitonin gene-related peptide (CGRP)-immunoreactive terminals in laminae I and II. With the demonstration of axosomatic, axodendritic as well as axoaxonic interactions, we hypothesize that CGRP-containing primary afferent terminals can effect processing at the level of the cell body (possibly influencing the manufacture of DYN and/or the firing rate of DYN-containing cells) and also modulate the output of DYN terminals in laminae I and II of the monkey dorsal horn. These structural relationships may play an important role in the processing of noxious input at the spinal cord level.

Animals↗
PubMed1990

CNS projections to the pterygopalatine parasympathetic preganglionic neurons in the rat: a retrograde transneuronal viral cell body labeling study.

The retrograde transneuronal viral cell body labeling method was used to study the CNS nuclei that innervate the parasympathetic preganglionic neurons which project to the pterygopalatine ganglion. Small injections of a suspension of pseudorabies virus (PRV) were made in the pterygopalatine ganglion of rats and after 4 days their brains wer e processed for immunohistochemical detection of PRV. Some of the tissues were stained with a dual immunofluoresence method that permitted the visualization of PRV and neurotransmitter enzyme or serotonin immunoreactivity in the same cell. Retrograde cell body labeling was detected in the ipsilateral ventrolateral medulla oblongata in the region that has been termed the superior salivatory nucleus. This area was the same region that was retrogradely labeled after Fluoro-Gold dye injections in the pterygopalatine ganglion. Retrograde transneuronally infected cell bodies that provide putative afferent inputs to the pytergopalatine parasympathetic preganglionic neurons were mapped throughout the brain. In the medulla oblongata, transneuronally labeled neurons were seen in the nucleus tractus solitarii, dorsomedial part of the spinal trigeminal nucleus and gigantocellular reticular nucleus. In most experiments, some A1 catecholamine cells and serotonin neurons of the raphe magnus, raphe pallidus, raphe obscurus, and parapyramidal nuclei were labeled. In the pons, labeled cells were found in the parabrachial nucleus. A5 catecholamine cell group, and non-catecholamine part of the subcoeruleus region. In the midbrain, cell body labeling was located in the central gray matter and retrorubral field. In the diencephalon, labeling was found mainly in the hypothalamus. The areas included the lateral hypothalamic area, lateral preoptic area, dorsomedial and paraventricular hypothalamic nuclei, and ventral zona incerta. Contralateral second order cell body labeling was seen in the tuberomammillary nucleus of the hypothalamus. Some of these cells were histidine decarboxylase-immunoreactive. In the forebrain, the bed nucleus of the stria terminalis, substantia innominata, and an area of the cerebral cortex called the amygdalopiriform transition zone were labeled.

Animals↗
PubMed1990

The cells of origin of the spinothalamic tract of the rat: a quantitative reexamination.

We quantitatively reinvestigated the cells of origin of the spinothalamic tract (STT) of the rat. Injections of Fluoro-Gold that filled the thalamus on one side labeled large numbers of neurons throughout the length of the spinal cord. In 3 cases, we estimated the total number of STT neurons by counting labeled neurons in 18 of the 34 total segments, applying correction factors to these counts, and estimating the numbers of labeled neurons in the 16 remaining unexamined segments. The accuracy of these estimates was tested in two animals in which labeled neurons were counted in all 34 spinal segments. In both cases, the estimated totals of STT neurons differed from the counted totals by less than 5%. In the most effective case, we estimated that more than 9500 STT neurons were labeled. This study indicates that the number of STT neurons in rats is larger than previously reported and suggests that the STT may play an important role in nociception in rats, as it does in primates including humans.

Animals↗