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PubMed1992

Thalamocortical connections of rat posterior parietal cortex.

The neuronal connections of rat posterior parietal cortex (PPC) have been examined using retrograde fluorescent axonal tracers. We have found that PPC receives thalamic input predominantly from the lateral posterior and lateral dorsal nuclei, and not from the ventrobasal nucleus, which projects to the rostrally adjacent hindlimb cortex, or from the dorsal lateral geniculate nucleus, which projects to the caudally adjacent visual association area. PPC has reciprocal corticocortical connections with medial agranular cortex and orbital cortex; together, these three cortical areas may function as a network for directed attention in rats.

Afferent Pathways↗
PubMed1992

Retrograde labelling of dorsal root ganglion cells in the rat: a quantitative and morphological comparison of Fluoro-Gold with horseradish peroxidase labelling.

We have compared retrograde labelling of rat primary sensory neurons using Fluoro-Gold (FG) and horseradish peroxide conjugated with wheat germ agglutinin (HRP-WGA). Fluoro-Gold 2.5% after 48 h transit time and FG 5% after 24 and 48 h retrogradely labelled similar numbers of cell profiles as HRP-WGA (P greater than 0.1% Student's t-test). The calculated cell size distribution for the above FG groups were similar to those for the HRP-WGA. However, FG 2.5% after a 24 h transit time labelled significantly fewer cells (P less than 0.001 Student's t-test). FG retrograde transport may be used to identify the same population of DRG cells as HRP-WGA.

Animals↗
PubMed1992

Identified septohippocampal neurons survive axotomy: a fine-structural analysis in the rat.

Previous studies have indicated that interruption of the connections between the medial septum and hippocampus by cutting the axons results in degeneration and death of the projecting septal neurons. However, in these studies cell death has been inferred primarily from the loss of immunoreactivity for transmitter-specific enzymes. In the present study, we labeled septohippocampal projection neurons by retrograde tracing and then cut their axons. Subsequent intracellular injection of prelabeled cells revealed the morphology of the soma and dendrites and allowed us to examine the ultrastructure of these neurons. A large number of septohippocampal neurons survived even 10 weeks after axotomy, suggesting that axotomized septohippocampal neurons survive for considerable periods beyond the time at which they stop expressing transmitter-specific immunoreactivity. Survival of axotomized neurons is a prerequisite for pharmacological interference aimed at reactivating transmitter expression, axonal re-growth, and the eventual reintegration into functionally relevant circuitries.

Animals↗
PubMed1992

NADPH diaphorase-positive neurons in the rat spinal cord include a subpopulation of autonomic preganglionic neurons.

Preganglionic neurons in the spinal cord of the rat were labelled retrogradely with Fluoro-gold and the spinal cord stained for NADPH diaphorase. The majority of both sympathetic and sacral parasympathetic preganglionic neurons showed staining for NADPH diaphorase. NADPH diaphorase-positive neurons were located more laterally in the intermediate zone than were preganglionic neurons lacking NADPH diaphorase staining. The recent evidence that identifies NADPH diaphorase as nitric oxide synthase raises the possibility that some spinal preganglionic neurons may synthesize nitric oxide.

3,3'-Diaminobenzidine↗
PubMed1992

Origin and distribution of NADPH-diaphorase-positive neurons and fibers innervating the urinary bladder of the rat.

Nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemistry was utilized to localize nitric oxide synthase (NOS), and thus sites where nitric oxide (NO) can be synthesized, within peripheral nervous system perikarya and fibers. Recent studies suggest that NO relaxes vascular and non-vascular smooth muscle. In this study, the origin and distribution of NADPH-diaphorase perikarya and fibers in the rat urinary bladder were examined. Results suggest that a small number of NADPH-diaphorase-positive perikarya are present within the bladder wall and within adjacent small ganglia. In addition, NADPH-diaphorase-positive nerve fibers were observed in the adventitial and muscular layers, subjacent to the urothelium and as perivascular fibers. After injection of the retrograde tracer fluorogold (FG) into the bladder wall, numerous FG-labeled perikarya in the major pelvic ganglia and the T13-L2, L6 and S1 dorsal root ganglia were NADPH-diaphorase positive. However, none of the FG-labeled perikarya in the inferior mesenteric ganglia were NADPH-diaphorase positive. The prevalence of NADPH-diaphorase-positive perikarya and fibers suggests that NO may serve a role in bladder function.

Amino Acid Oxidoreductases↗
PubMed1992

Connections of the thalamic reticular nucleus with the contralateral thalamus in the rat.

The occurrence of thalamic projections to the contralateral side of the thalamus, and the origin of these projections from the thalamic reticular nucleus, were demonstrated with fluorescent retrograde tracing. After a relatively large tracer injection in the anterior third of the thalamus (involving the reticular, anterior, intralaminar and ventrolateral nuclei), labeled cells were detected in the rostral portion of the contralateral reticular nucleus. The location within the reticular nucleus of the neurons retrogradely labeled in the contralateral thalamus was confirmed by means of the combination of tracing with immunocytochemistry, using antibodies against the calcium binding protein parvalbumin.

Animals↗
PubMed1993

Gonadotropin-releasing hormone (GnRH) neurons in the hypogonadal mouse elaborate normal projections despite their biosynthetic deficiency.

This study was undertaken to determine whether gonadotropin-releasing hormone (GnRH) neurons in the mutant hypogonadal (hpg) mouse can establish axonal connections with their target despite their failure to synthesize and secrete the GnRH neuropeptide. Normal and hpg males received intraperitoneal injections of the retrograde tracer Fluoro-Gold. This tracer does not cross the blood-brain-barrier and hence is taken up only by neurons in the central nervous system whose axons terminate on fenestrated capillaries, such as the capillaries of the median eminence. The brains of the injected animals were processed for in situ hybridization to visualize GnRH transcribing cells. In 3 hpg males 64.1 +/- 5.6% of GnRH transcribing cells contained Fluoro-Gold, while 55.8 +/- 6.4% of the cells in 3 normal males had Fluoro-Gold. Thus, we have demonstrated that secretory-deficient GnRH neurons can establish axonal connections with their primary secretory target, the median eminence. We conclude that the capability of GnRH neurons to recognize and interact with their target is not dependent upon their neurosecretory function.

Animals↗
PubMed1993

Localization of NADPH-diaphorase in pelvic afferent and efferent pathways of the rat.

The NADPH diaphorase histochemical reaction was used in combination with retrograde axonal transport of Fluorogold (FG) from the major pelvic ganglion (MPG) to determine if NADPH diaphorase is contained within afferent and preganglionic efferent pathways to pelvic visceral organs. In L6 and S1 dorsal root ganglia, 68.5% and 62.2%, respectively, of FG-labeled afferent neurons were NADPH-diaphorase positive. In the sacral parasympathetic nucleus (SPN) of the L6 and S1 spinal cord segments, 49.4% and 51.7%, respectively of FG labeled preganglionic efferent neurons were NADPH-diaphorase positive. NADPH-diaphorase-positive neurons were also observed in laminae I and II of the dorsal horn, around the central canal and in the dorsal commissure. In addition, fiber-like NADPH diaphorase staining was present in superficial dorsal horn, Lissauer's tract and the lateral edge of the dorsal horn extending into the region of the SPN. If NADPH diaphorase activity in neurons does indicate a physiological function of nitric oxide, then nitric oxide may have a role as a neuromodulator or neurotransmitter within visceral afferent and preganglionic efferent pathways to the pelvic viscera in the rat.

Afferent Pathways↗
PubMed1993

Rubrospinal neurons and retrograde transport of fluoro-gold in acute spinal cord injury--a dose-response curve.

Using the fluorescent tracer Fluoro-Gold, we studied the relationship between severity of spinal cord trauma and the number of retrogradely labeled rubrospinal neurons in the rat. We compared the mean cell counts of retrogradely labeled rubrospinal neurons in rats receiving 0, 20, 40, and 60 g/cm concussive spinal cord traumas. For each incremental increase in trauma a significant reduction in mean cell counts took place (P < 0.0001). We demonstrate a dose-response relationship between trauma severity and Fluoro-Gold retrogradely labeled rubrospinal neurons in acute spinal cord injury of the rat. This relationship may be helpful in quickly assessing the efficacy of therapeutic interventions in acute spinal cord injury. Previous studies with HRP failed to demonstrate such a dose-response curve. Fluoro-Gold may be a more sensitive indicator of neuronal survival than HRP in the traumatized rat spinal cord.

Acute Disease↗
PubMed1993

The major pelvic ganglion is the main source of nitric oxide synthase-containing nerve fibers in penile erectile tissue of the rat.

The possible implication of nitric oxide synthase (NOS) in penile erection was examined by utilizing NADPH histochemistry in the rat. NADPH histochemistry indicated that the major pelvic ganglion (MPG), a well-known origin of nerve fibers supplying the external genitalia, contained many NOS-positive neurons. On the other hand, NOS-positive nerve fibers in penile erectile tissue observed in the walls of both arteries and veins, as well as in intrinsic smooth muscles. The retrograde tracing study with Fluoro-Gold (FG) in combination with NADPH histochemistry revealed that almost all MPG neurons which were retrogradely labeled with FG injected into the penile crura were NOS-positive. Thus, the MPG was considered to be the main source of NOS-positive nerve fibers in penile erectile tissue.

Amino Acid Oxidoreductases↗
PubMed1993

Sensorimotor cortex projections to the ventrolateral and the dorsomedial medulla oblongata in the rat.

After small pressure injections of Fluorogold (FG), and Dextran-tetramethylrodamine (DR) into the dorsal motor nucleus of the vagus/nucleus of the solitary tract (DMV/NTS) and the rostral ventrolateral medulla (RVLM), respectively, retrograde FG-labelled cells were found mainly in the sensorimotor cortex; retrograde DR-labelled cells were located in the same cortical areas and in the prefrontal cortex. Double-labelled cells were also found in the sensorimotor cortical areas. These results provide evidence of direct projections from the sensorimotor cortex to the DMV/NTS and RVLM and suggest that somatic cortical areas directly control cardiovascular output during sensory and somatic processes.

Animals↗
PubMed1993

NADPH diaphorase innervation of the rat anococcygeus and retractor penis muscles.

NADPH diaphorase histochemistry was used to determine whether the rat anoocccygeus (AC) and retractor penis (RP) muscles are innervated by nerves capable of synthesizing nitric oxide. In both tissues, muscle fascicles were enclosed by a varicose plexus of NADPH diaphorase positive (ND+) fibers. Perikarya of neurons on the surface of the AC muscle were also intensely stained for NADPH diaphorase. Many AC-RP ganglion cells in the pelvic plexus, located by the retrograde tracer Fluorogold, also stained for the enzyme. However, a significant population of AC-RP neurons in this location remained unstained. These results provide further evidence that nitric oxide may be an important neurotransmitter in these tissues.

Animals↗
PubMed1994

NADPH-diaphorase-positive neurons innervating the rat ovary.

Nerve cell bodies projecting to the ovary were visualized in dorsal root ganglia (DRG) and paravertebral ganglia after application of the retrograde tracer Fluoro-gold to the superior ovarian and plexus nerves. The location of fluorescent cells in sections of ganglia was recorded and subsequently nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemistry was utilized to locate nitric oxide synthase (NOS) whose presence indicates sites where nitric oxide (NO) can be synthesized. No fluorescent nerve cell bodies in paravertebral ganglia were NADPH-diaphorase-positive. In contrast, numerous Fluoro-gold-labeled nerve cell bodies in DRG at segmental levels T12-L1 were NADPH-diaphorase positive. Thus, many sensory neurons projecting to the ovary contain NOS and presumably release NO. This gaseous messenger molecule may participate in modulation of ovarian function.

Afferent Pathways↗
PubMed1994

Immunocytochemical co-localization of substance P and calcitonin gene-related peptide in afferent renal nerve soma of the rat.

Substance P, calcitonin gene-related peptide and somatostatin immunoreactivities have been demonstrated in putative afferent renal nerve fibers in the rat. Utilizing retrograde-tracing and immunohistochemistry, we labeled afferent renal nerve soma throughout dorsal root ganglia T9 to L1. Most (85%) of afferent renal nerve perikarya were immunoreactive for calcitonin gene-related peptide, 21% had substance P immunoreactivity and none had somatostatin immunoreactivity. All renal afferents immunoreactive for substance P also contained calcitonin gene-related peptide. These results provide evidence that calcitonin gene-related peptide and substance P are present and co-localized in afferent renal nerves, and therefore, mediate transmission of afferent renal input to the spinal cord in the rat.

Amidines↗
PubMed1994

Reciprocal connections of lateral septal neurons and neurons in the lateral hypothalamus in the rat: a combined phaseolus vulgaris-leucoagglutinin and Fluoro-Gold immunocytochemical study.

Reciprocal connections between lateral septal neurons and neurons in the lateral hypothalamus/lateral preoptic area were studied in the rat. The anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L) and the retrograde tracer Fluoro-Gold (FG) were simultaneously injected into the lateral septum. After double-immunocytochemistry, PHA-L-labeled terminals were found in synaptic contact with dendrites of retrogradely FG-labeled neurons in the lateral hypothalamic/lateral preoptic area.

Afferent Pathways↗
PubMed1994

The consequence of delayed versus immediate nerve repair on the properties of regenerating sensory nerve fibres in the adult rat.

Transected saphenous neurones were allowed to regenerate for 3 months via distal stumps of sural nerve following an immediate or a 3 month delayed repair. The number of DRG neurons surviving following the 3 months regeneration period were approximately 60% of normal after both immediate and delayed repair. The percentage of DRG cell bodies identified by the application of Fluro-gold proximal to the repair site and immunopositive for SP, CGRP and galanin was increased following both early and delayed repair compared to baseline values. These values were not significantly different for early repair compared to late repair. Similarly, peripheral measurements of SP in the proximal stump of saphenous nerve (by radioimmunoassay) were not significantly different between models with primary repair compared to delayed repair. These results suggest that the intrinsic regeneration properties of primary sensory neurones are not impaired when repair is delayed.

Anastomosis, Surgical↗
PubMed1994

Thalamocortical projections activated by phrenic nerve afferents in the cat.

This study identified thalamocortical projections activated by respiratory afferents. Cortical evoked potentials were recorded in the right primary somatosensory cortex of the cat following electrical stimulation of the left C5 root of the phrenic nerve. The majority of primary sites were located in the vicinity of the postcruciate dimple, in area 3a near the 3a/3b border, corresponding to the trunk region of the cortical body map. Retrograde fluorescent tracers injected at the sites of primary activation produced labeled cells in the oralis nucleus of the ventroposterior complex [4]. Control injections made in adjacent cortical areas not activated by phrenic stimulation resulted in labeling in the ventroposterior complex which did not overlap that seen with injections of primary activation sites. We conclude that respiratory muscle afferents in the phrenic nerve elicit activity in the trunk region of primary somatosensory cortex via specific thalamocortical projections originating in the oralis portion of the thalamic ventroposterior complex.

Afferent Pathways↗
PubMed1994

Basic fibroblast growth factor-like immunoreactivity in the rat basilar artery with reference to co-localization with NADPH-diaphorase in the trigeminal ganglion.

The location of basic fibroblast growth factor (bFGF)-like immunoreactivity and nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase (neuronal nitric oxide synthase) activity in the rat basilar artery and in the trigeminal, sphenopalatine and superior cervical ganglia was investigated. bFGF immunoreactivity was seen mainly in adventitial nerve fibers of the rat basilar artery, but not in the endothelium. Electron microscopy of the tunica media showed a number of immunoreactive nerve endings in the vicinity of local smooth muscle cells. Among the cranial ganglia that innervate the basilar artery, only the trigeminal ganglion had bFGF-immunoreactivity neurons. Nerve cells and fibers with NADPH-diaphorase activity were detected in the basilar artery and in the sphenopalatine and trigeminal ganglia, and the co-localization of bFGF and NADPH-diaphorase was noted only in the trigeminal ganglion. Furthermore, Fluro-gold tracing in combination with bFGF immunohistochemistry demonstrated that bFGF-containing nerve fibers in the wall of the basilar artery arise from the trigeminal ganglion. These findings provide a morphological basis for the nitric oxide-mediated dilatation of cerebral arteries by bFGF.

Animals↗