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The hypothalamus receives major projections from the tuberomammillary nucleus in rat.

Projections from the tuberomammillary nucleus (TM) to widespread regions within the hypothalamus were evaluated using a combined immunohistochemical-retrograde fluorescent tracing procedure. Injections of Fluoro-gold into the anterior hypothalamus labelled TM neurons identified by their immunoreactivity for the enzyme adenosine deaminase (ADA). Small injections of Fluoro-gold into the posterior hypothalamus also led to the labelling of TM neurons. The numbers of ADA-immunoreactive axonal varicosities were 5 times greater in the hypothalamus than in most other brain regions. The results indicate that the hypothalamus represents a major projection area of the TM.

Adenosine Deaminase↗

A direct projection from the tuberomammillary nucleus to the spinal cord in the rat.

After injections of retrograde fluorescent tracers (True blue (TB), Diamidino yellow (DY) and Fluorogold) into the upper cervical segments of the rat spinal cord, a group of labeled neurons were consistently found bilaterally in the tuberomammillary nucleus (TM) of the hypothalamus with an ipsilateral predominance. The data from injections of two different tracers (TB and DY) in the same animal suggest that the vast majority of TM cells projecting to the spinal cord rarely give off divergent axon collaterals to the frontal/prefrontal cortices, striatum, amygdala and superior colliculus.

Amidines↗

A double-label study of efferent projections from the Edinger-Westphal nucleus in goldfish and kelp bass.

The Edinger-Westphal nucleus in goldfish was identified by retrograde labeling from the ciliary ganglion. In the same animals a few neurons near this nucleus (perinuclear Edinger-Westphal neurons) were labeled by a different retrograde tracer injected into the cerebellum. No double-labeled cells were found. Similar results were obtained in kelp bass, except that in this species no cerebellar-projecting perinuclear neurons were observed. Cerebellar-projecting Edinger-Westphal neurons have previously been described in some mammals, but not in other vertebrates. Therefore the homology of cerebellar-projecting cells of the Edinger-Westphal region in mammals and teleost fishes is doubtful.

Animals↗

Location of motoneurons and internuclear neurons within the rat abducens nucleus by means of horseradish peroxidase and fluorescent double labeling.

The distribution of abducens motoneurons and internuclear neurons was determined in the rat by injections of horseradish peroxidase or fluorochromes into the ipsilateral lateral rectus muscle and the contralateral oculomotor nucleus either separately or simultaneously. The labeled somata of abducens internuclear neurons were intermingled with the labeled motoneurons at the medial third of the nucleus, but they were more segregated at the rostral third, where the labeled interneurons were more numerous. Internuclear neurons were preferentially located around and ventral to the central part of the facial genu, while motoneurons were located more dorsomedially, closer to the midline than in other species of mammals. The evolutionary trend of the location of both populations of neurons is also discussed.

Abducens Nerve↗

Non-cholinergic basal forebrain neurons project to the contralateral basal forebrain in the rat.

Following injections of wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) or the fluorescent tracer fluoro-gold into the magnocellular preoptic area and the horizontal limb of the diagonal band, retrogradely labelled neurons were found in the homotopic region of the contralateral basal forebrain. Labelled fibers apparently arising from these neurons travelled in the stria medullaris and the habenular commissure to terminate in the contralateral basal forebrain. Although the neurons retrogradely labelled with fluoro-gold in the contralateral basal forebrain were similar in size to choline acetyltransferase (ChAT)-immunoreactive neurons, and were intermingled with them, none was ChAT-positive. WGA-HRP injections into the nucleus basalis magnocellularis did not result in retrograde labelling in the contralateral basal forebrain. These findings suggest that non-cholinergic neurons may serve as a direct link between the two sides of selective magnocellular basal forebrain regions.

Animals↗

Adrenergic and non-adrenergic neurons in the C1 and C3 areas project to locus coeruleus: a fluorescent double labeling study.

Following iontophoretic injections of the retrograde tracer Fluoro-gold into the rat locus coeruleus (LC), retrogradely labeled neurons were seen predominantly in the area of C1 adrenergic neurons in the ventrolateral medulla (nucleus paragigantocellularis; PGi) and in the area of C3 adrenergic neurons in the dorsomedial medulla (nucleus prepositus hypoglossi; PrH). Subsequent immunofluorescence for phenylethanolamine-N-methyltransferase indicated that adrenergic and non-adrenergic LC projecting neurons in both areas are interdigitated, and that 21% of LC afferent neurons in the PGi are adrenergic while only 4% of LC afferent neurons in the area of PrH are adrenergic.

Adrenergic Fibers↗

Intraretinal transplantation of fluorescently labeled retinal cell suspensions.

Dissociated cell suspensions of neonatal neural retina, labeled with the fluorescent dyes Fast blue or Fluoro-gold, were transplanted into the retina of normal adult rats or of rats affected by late stage phototoxic retinopathy. Light microscopy showed good survival, differentiation, and integration of the transplants, as well as permanence of the label up to 100 days. The results indicate that the transplantation of dissociated, fluorescently labeled retinal cells has a number of advantages over the transplantation of solid fragments of retinal tissue, previously performed by ourselves and others. The following are some of the most immediate procedural advantages: the number of transplanted cells can be assessed, the transplanted cells are in a more intimate contact with the host tissue and therefore integrate better with the host, and the fluorescent tags permit precise determination of the survival and distribution of the transplanted cells.

Amidines↗

Fluoro-Gold reveals patches of striatal neurons during development.

Patches of densely-labelled striatal neurons were observed following intrastriatal injection of the fluorescent dye Fluoro-Gold into rats younger than 10 days of age. These cell clusters showed precise overlap with islands of dense dopamine innervation seen only during the first week of life. The patches of labelled cells were still visible in adulthood, and were not abolished by 6-hydroxydopamine treatment before or after the Fluoro-Gold injection. Neonatal Fluoro-Gold injection may be a unique way of marking the patch compartment of the striatum that remains visible in adulthood.

Aging↗

Layer VII of rodent cerebral cortex.

The cerebral isocortex is usually considered to be a 6-layered structure. Our anatomical findings suggest that layer VII be recognized as a distinct entity in rodent isocortex. This conclusion is based on cytoarchitectural, fiberarchitectural, connectional and developmental data.

Animals↗

Parvalbumin immunoreactivity in the rat retina.

The distribution of the Ca2+ binding protein parvalbumin was studied in the rat retina with immunocytochemistry using a mouse monoclonal antibody. Specific parvalbumin immunoreactivity was identified within a subpopulation of ganglion cells and a subpopulation of amacrine cells. The topographical data provided by the present study may serve as a basis for a functional characterization of parvalbumin's role in the nervous system.

Animals↗

Photoconversion of fluorescent retrograde tracers.

Photoconversion of fluorescent dyes, retrogradely transported through axons to their parent cell bodies, into a stable diaminobenzidine (DAB) reaction product was tested in the nigrostriatal and thalamocortical systems of rats. Satisfactory results were obtained with Propidium Iodide (PI), Fluoro-Gold (FG), Fast Blue (FB), Diamidino Yellow (DY), and rhodamine-labeled latex microspheres (RLM); some photoconversion was also observed in Evans Blue (EB)-labeled neurons. The red fluorescent tracers PI, EB and RLM were photoconverted under the excitation wavelength appropriate for eliciting their fluorescent emission. With the yellow or blue fluorescent tracers FG, FB, and DY satisfactory results could instead be obtained using an excitation wavelength which did not elicit visible fluorescent emission. This finding indicates that the latter is not a critical factor for obtaining photoconversion. The features and subcellular localization of photoconverted DAB were different from those of the fluorescent labeling: photoconversion resulted in the appearance of brown granules of DAB reaction products in the cytoplasm, independently from the occurrence of fluorescent labeling in the neuronal cytoplasm or nucleus. Photoconversion may enable new applications of fluorescent retrograde tracing and, in particular, its electron microscopic visualization.

Animals↗

The noradrenergic innervation of the spinal cord: differences between two substrains of Sprague-Dawley rats determined using retrograde tracers combined with immunocytochemistry.

We have recently described the spinal cord terminations of noradrenergic neurons located in the A5, A6 and A7 cell groups. However, recent reports from another laboratory, using similar experimental methods, have described results that are profoundly different. The present experiments were designed to determine whether these discrepant results are due to fundamental differences between the substrains of rats used in the conflicting experiments. To this end, retrograde tract tracing experiments were done using Sprague-Dawley rats from either Sasco, Inc. or Harlan Sprague-Dawley, Inc. The results indicate that noradrenergic neurons in the pontine catecholamine cell groups exhibit remarkably different spinal cord projections in these two substrains of Sprague-Dawley derived rats.

Animals↗

Cholinergic projections from the globus pallidus to the lateral amygdaloid nucleus in the cat.

Direct projections from the globus pallidus (GP) to the lateral amygdaloid nucleus (A1) were found in the cat by the anterograde and retrograde tracing methods. Pallido-amygdaloid fibers originated mainly from the caudal half of the GP and distributed throughout the entire extent of the A1; most densely in the lateral part of the A1. Choline acetyltransferase immunohistochemistry in combination with retrograde fluorescent tracing indicated that most pallido-amygdaloid neurons were cholinergic.

Amygdala↗

Transmitter synthesis increases in substantia nigra neurons of the aged mouse.

Striatal dopamine concentrations are relatively well maintained with age despite extensive death of the nigrostriatal neurons whose terminals contain the dopamine. Counts of nigrostriatal dopaminergic neurons in C57BL mice identified using immunocytochemistry, Fluoro-Gold retrograde axonal transport and Nissl staining were combined with measures of striatal dopamine and DOPA after saline, pargyline or NSD-1015 treatment. On average, 68% of the dopaminergic nigrostriatal neurons died between ages 8 and 104 weeks and there was a 3-fold increase in dopamine synthesis per average neuron in the aged mice. Increased transmitter synthesis by surviving neurons may serve to compensate brain function in old age.

3,4-Dihydroxyphenylacetic Acid↗

Light-dependent development and asymmetry of visual projections.

The projections from thalamus to visual Wulst in chicks are asymmetrical and their development is determined by exposure to light just before and after hatching. The asymmetry results from the orientation of the embryo in the egg, the left eye being occluded and not the right. We have shown that this asymmetry can be eliminated by incubating the embryos in darkness or light so that both eyes receive the same stimulation.

Animals↗

Non-dopaminergic projection from the subparafascicular area to the temporal cortex in the rat.

Following injection of fluorescent tracer into the rat temporal cortex, retrogradely labeled cells were found in the subparafascicular area. In the thalamus, numerous cell labeling was seen in the medial geniculate complex. Labeled cells in the subparafascicular area were small (10-20 microns), and did not display tyrosine hydroxylase immunoreactivity. From these findings, the subparafascicular area has, at least, two distinct projection systems towards the cerebral cortex; the previously described large dopaminergic cells innervating the frontal cortex and small non-dopaminergic cells innervating the temporal cortex.

Animals↗

Evidence for a projection from the globus pallidus to the entopeduncular nucleus in the rat.

Iontophoretic injections of the fluorescent retrograde tract tracer, Fluoro-gold, into the entopeduncular nucleus of the rat (homologous to the internal segment of the primate globus pallidus) resulted in a substantial number of retrogradely labeled neurons in the ipsilateral globus pallidus (homologous to the external segment of the primate globus pallidus). In experiments confirming this projection, iontophoretic injections of the anterograde tract tracer, Phaseolus vulgaris-leucoagglutinin, in the globus pallidus resulted in dense fiber and terminal labeling in the ipsilateral entopeduncular nucleus. This projection is topographically organized in rostral-caudal, medial-lateral and dorsal-ventral orientations.

Animals↗

Fluoro-Gold's toxicity makes it inferior to True Blue for long-term studies of dorsal root ganglion neurons and motoneurons.

We studied the long-term effects of two retrogradely transported fluorescent dyes on survival of dorsal root ganglion neurons (DRGNs) and motoneurons (MNs). In adult female rats, we labeled DRGNs and MNs by soaking the cut sciatic nerve in Fluoro-Gold or True Blue. With True Blue, we found no difference in the number of labeled MNs or DRGNs in rats surviving 4 days or 20 weeks after nerve soak. With Fluoro-Gold, labeled DRGNs and MNs were decreased at 20 weeks compared with 4 days. Since there was no offsetting increase in unlabeled DRGNs at 20 weeks, Fluoro-Gold caused cell death.

Animals↗