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Query: EC:1.11.1.7 (peroxidase)
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The central distribution of visceral primary afferent fibers from the pelvic nerve of the cast and the relationship of these fibers to preganglionic neurons of the sacral parasympathetic neurons (SPN) have been studied. Horseradish peroxidase (HRP) applied to the cut pelvic nerve was detected ipsilaterally in preganglionic neurons and dorsal root ganglion cells (segments S1-S3), and in central afferent projections to Lissauer's tract (LT), the dorsal columns, the dorsolateral funiculus, and spinal gray matter. The afferent projections were strongest in the region of the SPN (S1-S3) but extended far beyond its limits (e.g., LT was labeled from L4 to Cx7). In the transverse plane, collateral fiber bundles formed a thin shell around the dorsal horn predominantly within lamina I and expanded into terminal fields in the gray matter. The more prominent lateral collateral projection (LCP) extended into laminae V and VI, whereas the medial one (MCP) ended in the dorsal commissure. In longitudinal planes these projections exhibited a periodicity with an interval of approximately 200 micrometer. The distribution of afferent collateral projections overlaps the regions where many preganglionic neurons and their dendritic extensions are located, and also areas known to contain interneurons involved in visceral pathways. A differential distribution of afferents within the SPN was noted where a higher intensity was observed in proximity to those neurons located in laminae V and VI, which innervate the colon, and a lower intensity near neurons located in Lamina VII which innervate the bladder. This is consistent with the known spinal control of colon reflexes and the supraspinal control of bladder reflexes. The widespread rostrocaudal extent of the pelvic primary afferent projection is consistent with the necessity for the integration of somatic and autonomic elements from various levels of the lumbo-sacral-coccygeal spinal cord in the performance of pelvic visceral functions.
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PMID:The distribution of visceral primary afferents from the pelvic nerve to Lissauer's tract and the spinal gray matter and its relationship to the sacral parasympathetic nucleus. 727 58

The distribution of afferents innervating the urinary bladder in the spinal cord of male rats has been studied with the axonal tracer horseradish peroxidase conjugated to wheat germ agglutinin (WGA-HRP) injected into various portions of one side of the urinary bladder (dome, body, base, or neck) and other pelvic organs (prostate and rectum). Labeled neurons were found in dorsal root ganglia of the lumbosacral cord (L1-S3, peak in S1-S2). The strongest and most extensive transganglionic labeling of primary afferents resulted after injections in the body of the bladder. Primary afferents were observed bilaterally in Lissauer's tract and laminae I-II at the apex of the dorsal horn, from L6 to S3. The projection extended laterally up to the sacral parasympathetic nucleus and medially up to the gray matter dorsal to the central canal, where they formed a plexus of fibers and terminals. Deposits in the dome and base of the bladder labeled more heavily the medial projection, while the least intense projection was seen after injections in the bladder neck. Our results indicate a common pattern of termination of primary afferents from the bladder, although some topographical differences exist.
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PMID:Urinary bladder innervation in male rat: termination of primary afferents in the spinal cord as determined by transganglionic transport of WGA-HRP. 768 86

The projections of the ankle joint capsule afferents were studied by transganglionic transport of horseradish peroxidase injected directly into the ankle joint. The number and size of the labelled dorsal root ganglion cells were measured from synsacral nerves 2-9. In the dorsal root ganglia, all sizes of sensory neurones were labelled, and the largest number of labelled cells was in ganglia 5-7. The extensive sympathetic innervation of the ankle joint was identified by the large number of cell bodies labelled in the sympathetic ganglia of the paravertebral chain. Labelled afferent fibres projected to the spinal cord from the 2nd to the 8th synsacral nerves, with the rostral projection mainly via Lissauer's tract and the dorsal funiculus. Terminal labelling in the dorsal horn was identified in laminae I-III and VI, with a slight projection to V. Two areas of dense labelling, which did not correspond with the largest number of labelled dorsal root ganglion cells, were identified. A rostral area with the highest density of label was observed at the level of synsacral nerves 3-4 and a second slightly less dense area between synsacral nerves 7-8. In the caudal medulla, diffuse terminal labelling was observed in the nucleus gracilis et cuneatus, nucleus of the tractus solitarius, and the nucleus cuneatus externus. These results are discussed in a comparative context to identify similarities and differences between different primary afferent projections in birds and mammals and to highlight the possible functional significance of the avian articular afferent projection.
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PMID:Projections of ankle joint afferents to the spinal cord and brainstem of the chicken (Gallus g. domesticus). 857 21

Injections of cholera toxin B-chain conjugated to horseradish peroxidase into individual peripheral branches of the trigeminal nerve or into the trigeminal ganglion showed that an ascending trigeminal tract (TTA) terminated in distinct ventral and dorsal divisions of the principal sensory nucleus (PrVv and PrVd, respectively), and a descending tract (TTD) terminated within pars oralis, pars interpolaris, and pars caudalis divisions of the nucleus of TTD (nTTD) and within the dorsal horn of the first six cervical spinal segments. In PrVd, mandibular, ophthalmic, and maxillary projections were predominantly located dorsally, ventrally, and medially, respectively. In nTTD, mandibular projections lay dorsomedially, ophthalmic projections lay ventrolaterally, and maxillary projections lay in between. At caudal medullary and spinal levels, mandibular projections were situated medially, ophthalmic projections were situated laterally, and maxillary projections were situated centrally. The terminations within the dorsal horn were most dense in laminae III and IV and were least dense in lamina II, with laminae III-IV also receiving topographically organised contralateral projections. Extratrigeminal projections were mainly to the external cuneate nucleus by way of a lateral descending trigeminal tract (lTTD; Dubbeldam and Karten [1978] J. Comp. Neurol. 180:661-678) and to the region of the tract of Lissauer and lamina I of the dorsal horn. Other projections were to a region medial to the apex of pars interpolaris, to the nuclei ventrolateralis anterior (Vla) and presulcalis anterior (Pas) of the solitary complex, and sparsely to the lateral reticular formation (plexus of Horsley) ventral to TTD. No projections were seen to the trigeminal motor nuclei or to the cerebellum.
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PMID:Central projections and somatotopic organisation of trigeminal primary afferents in pigeon (Columba livia). 872 98

We have studied the cyto- and myeloarchitectural organisation of the spinal cord of an echidna (Tachyglossus aculeatus) with the aid of Nisst staining, darkfield examination and p-phenylenediamine staining. We have also examined the distribution of unmyelinated afferents by labelling with a peroxidase-conjugated lectin derived from Griffonia simplicifolia (B1 isolectin). The cytoarchitectural features characterising the laminar organisation of the spinal cord in eutherian mammals were broadly applicable to the spinal cord of this monotreme. In addition, we identified a distinct group of large neurons in the ventral part of lamina X, extending into the ventral funiculus, that we have called the median nuclear group. We were unable to identify a central cervical nucleus in this echidna on the basis of cytoarchitectural criteria, although all other spinal cord nuclei found in eutherians could be found in this monotreme. Lectin labelling with the Griffonia simplicifolia isolectin B4 revealed a subpopulation of dorsal root ganglion cells similar to those labelled in Eutheria. In this echidna, labelling of unmyelinated fibres was found in Lissauer's zone and laminae I and II, as seen in rats (Rattus norvegicus); there were also deeper patches extending into laminae III to V and what appeared to be commissural axons approaching the dorsal grey commissure, which have not been seen in Eutheria. Fibre calibre in the dorsal and ventral roots of this echidna was similar to that reported in Eutheria, suggesting similar proportion of afferent fibre classes and alpha and gamma motoneurons. In the echidna, mean diameter of myelinated dorsal root axons was 4.65 microns at T1 and 5.22 microns at L3, with a clear bimodal distribution in the L3 dorsal root showing distinct groups at 1 to 5 microns and 6 to 12 microns. These made up approximately 45 and 55% of the total myelinated axon population, respectively. Myelinated fibres in the ventral root at L3 showed two major peaks in distribution. These were at 1 to 4 microns (approximately 32% of the total myelinated fibre population) and at 7 to 14 microns (approximately 58% of the total myelinated fibre population). The cross-sectional area of the dorsal columns of this monotreme was comparable to that of a eutherian mammal of similar body weight, and myelinated axon calibre was similar to that seen in the domestic cat. Our findings indicated that spinal cord cytoarchitectural organisation is highly conserved across class Mammalia, although the lectin labelling findings suggested that termination of unmyelinated afferents in echidnas may differ from that found in Eutheria. The dorsal column system appears to be as anatomically well developed in this monotreme as in those eutherian mammals considered to have a pronounced discriminative tactile sense.
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PMID:Cyto- and myeloarchitectonic organisation of the spinal cord of an echidna (Tachyglossus aculeatus). 915 Aug 97

In the present study, we investigated and compared the ability of the cholera toxin B subunit, wheat germ agglutinin and isolectin B4 from Griffonia simplicifolia I conjugated to horseradish peroxidase, to retrogradely and transganglionically label visceral primary afferents after unilateral injections into the rat urinary bladder wall. Horseradish peroxidase histochemical or lectin-immunofluorescence histochemical labelling of bladder afferents was seen in the L6-S1 spinal cord segments and in the T13-L2 and L6-S1 dorsal root ganglia. In the lumbosacral spinal cord, the most intense and extensive labelling of bladder afferents was seen when cholera toxin B subunit-horseradish peroxidase was injected. Cholera toxin B subunit-horseradish peroxidase-labelled fibres were found in Lissauer's tract, its lateral and medial collateral projections, and laminae I and IV-VI of the spinal gray matter. Labelled fibres were numerous in the lateral collateral projection and extended into the spinal parasympathetic nucleus. Labelling from both the lateral and medial projections extended into the dorsal grey commissural region. Wheat germ agglutinin-horseradish peroxidase labelling produced a similar pattern but was not as dense and extensive as that of cholera toxin B subunit-horseradish peroxidase. The isolectin B4 from Griffonia simplicifolia I-horseradish peroxidase-labelled fibres, on the other hand, were fewer and only observed in the lateral collateral projection and occasionally in lamina I. Cell profile counts showed that a larger number of dorsal root ganglion cells were labelled with cholera toxin B subunit-horseradish peroxidase than with wheat germ agglutinin- or isolectin B4-horseradish peroxidase. In the L6-S1 dorsal root ganglia, the majority (81%) of the cholera toxin B subunit-, and almost all of the wheat germ agglutinin- and isolectin B4-immunoreactive cells were RT97-negative (an anti-neurofilament antibody that labels dorsal root ganglion neurons with myelinated fibres). Double labelling with other neuronal markers showed that 71%, 43% and 36% of the cholera toxin B subunit-immunoreactive cells were calcitonin gene-related peptide-, isolectin B4-binding- and substance P-positive, respectively. A few cholera toxin B subunit cells showed galanin-immunoreactivity, but none were somatostatin-, vasoactive intestinal polypeptide-, or neuropeptide Y-immunoreactive or contained fluoride-resistant acid phosphatase. The results show that cholera toxin B subunit-horseradish peroxidase is a more effective retrograde and transganglionic tracer for pelvic primary afferents from the urinary bladder than wheat germ agglutinin-horseradish peroxidase and isolectin B4-horseradish peroxidase, but in contrast to somatic nerves, it is transported mainly by unmyelinated fibres in the visceral afferents.
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PMID:Retrograde and transganglionic transport of horseradish peroxidase-conjugated cholera toxin B subunit, wheatgerm agglutinin and isolectin B4 from Griffonia simplicifolia I in primary afferent neurons innervating the rat urinary bladder. 972 57

A modification of the horseradish peroxidase method was used to stain selectively dorsal root fibers entering the spinal cord of the cat. This technique stains intact terminal arborizations. Using this method to examine the primary afferent supply to lamina II of the lumbosacral segments, we observed that both thick primary afferent collaterals (2-3 mum), entering by way of the dorsal column, and thinner primary afferent collaterals (<1 mum), entering by way of Lissauer's tract, form terminals in lamina II. The terminals derived from the thick primary afferent collaterals were concentrated in the ventral portion of lamina II.
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PMID:Staining of the dorsal root projection to the cat's dorsal horn by anterograde movement of horseradish peroxidase. 1960 78

Sacral afferent and preganglionic efferent neurons were labelled by applying horseradish peroxidase (HRP) to the central end of the transected pelvic nerve. Preganglionic cells were evident as a narrow band of cells extending from the intermediolateral region dorsomedially beneath the dorsal horn. Their dendrites were located primarily within the nucleus but also projected into the dorsolateral funiculus, along the lateral edge of the dorsal horn and to dorsal commissure. Labelled primary afferent axons and/or terminals were detected in Lissauer's tract and in the marginal zone of the dorsal horn extending from the apex ventrally along its lateral edge into the dorsal region of the autonomic nucleus. These observations suggest that sacral preganglionic neurons may receive monosynaptic connections from pelvic nerve afferents.
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PMID:Horseradish peroxidase tracing of visceral efferent and primary afferent pathways in the cat's sacral spinal cord using benzidine processing. 1960 64


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