Distribution of substance P-like immunoreactivity in the lower brainstem of the human fetus: An immunohistochemical study

The regional distribution of substance P-like immunoreactive (SPI) structures in the lower brainstem of the human fetus was investigated using the indirect immunoflourescence technique. SPI cells were found in a number of areas including the inferior colliculus, central gray matter of the midbrain, n. laterodorsalis tegmenti, midbrain and medullary reticular formation, n. vestibularis inferior, and n. prepositus hypoglossi. An extensive network of SPI fibers of varying densities were identified throughout the lower brainstem.     

Distribution and quantitative measurements of somatostatin-like immunoreactivity in the lower brainstem of the rat

A radioimmunoassay was used to identify and quantify somatostatin-like immunoreactivity in the lower brainstem of the rat. The peptide was detectable in all brainstem nuclei investigated. Highest levels were found in the nuclei parabrachiales, central gray, ocus coeruleus, and nucleus tractus solitarii. Extremely low levels were found in the substantia nigra and cerebellum.     

Axonal transport of substance P-like immunoreactivity in regenerating rat sciatic nerve

Anterograde axonal transport of substance P-like immunoreactivity (SPLI) decreases after injury (crush or resection) to rat sciatic nerve. If the axons regenerate a partial recovery of transport occurs. If regeneration is impeded the decrease in transport is more severe and prolonged. No changes in the proportion of mobile SPLI (31%) or transport velocity (10.0 mm/h) occur. The decrease in SPLI transport largely accounts for the decline in SPLI content which occurs in nerve following injury and probably reflects decreased cell body synthesis.     

Regional distribution of methionine-enkephalin and substance P-like immunoreactivity in normal human brain and in Huntington''s disease

The regional distributions of substance P and Methionine-enkephalin (Met-enkephalin) were determined in normal human brains and in Huntington's disease using sensitive radioimmunoassays. Model experiments showed that both Met-enkephalin- and substance P-like immunoreactivities were stable for up to 72 h post-mortem in mouse brain. The results of high pressure liquid chromatography (HPLC) analyses indicated that the majority of the immunoreactivity detected in human globus pallidus corresponded to the native peptides, substance P or Met-enkephalin. In Huntington's disease the present results confirm that there is a substantial drop (> 80%) in the substance P content of the globus pallidus (both medial and lateral segments) and substantia nigra, and there was also a reduction (> 50%) in the Met-enkephalin content of these areas. This result suggests the loss of striato-pallidal and striato-nigral substance P and enkephalin-containing projections in Huntington's disease.     

Chemical characterization of substance P-like immunoreactivity in primary afferent neurones

Substance P-like immunoreactivity (SPLI) in rat dorsal root ganglia and dorsal spinal cord was characterized by high-performance liquid chromatography followed by radioimmunoassay. In spinal cord and ganglia, respectively, 87% and 64% of SPLI eluted with authentic SP. The remainder of the SPLI in ganglia eluted as a single peak which did not represent the sulphoxide of SP or any of its C-terminal fragments.     

Localization of immunoreactive thyrotropin releasing hormone in the lower brainstem of the rat

The distribution of thyrotropin releasing hormone (TRH) in individual nuclei of the rat brainstem was examined by radioimmunoassay. TRH was detectable in 36 of 40 brainstem nuclei investigated and the localization of TRH in the brainstem was unlike other known brainstem neuropeptides. By far the highest concentration of TRH in the brainstem (1.2 ng/mg protein) is present in the nucleus of the solitary tract. The concentration of TRH was relatively high in the motor nuclei of the IIIrd, Vth, VIIth, Xth and XIIth cranial nerves, and less high in the area postrema, nucleus gracilis, locus coeruleus, lateral reticular nucleus (A1-catecholamine cell group), dorsal raphe and central gray matter. Cerebellum and pontine nuclei contained very low levels (<0.03ng/mg protein) of TRH.     

Regeneration of primary afferent neurons containing substance P-like immunoreactivity

We compared changes in levels of substance P-like immunoreactivity (SPLI) in L4-6 dorsal root ganglia (DRG), L4-6 dorsal roots, sciatic nerve, tibial nerve and hind foot skin in rats following resection or crush injury of the sciatic nerve. The initial depletion of SPLI, which occurred in all areas sampled, was similar after either type of lesion. In DRG and dorsal roots, recovery to control values occurred in SPLI levels 35-45 days after sciatic crush, but not after resection. In sciatic nerve proximal to the injury, a partial recovery in SPLI content to about 60% of control occurred following crush injury, but not following resection. Distal to the injury, tibial nerve levels recovered rapidly following crush injury, consistent with the previously observed rapid regeneration of SPLI-containing axons. After resection, no recovery was observed until after 35 days, when it appeared that some axons succeeded in crossing the resection zone and regaining the distal nerve stump. Delayed and poor recovery of SPLI levels was observed in foot skin, even after crush injury. This correlated with the poor recovery of the plasma extravasation reaction, a functional index of SP-innervation of skin. In contrast, reinnervation by high-threshold mechanoreceptors was more rapid and complete, in agreement with a previous study. We conclude that although SPLI-containing axons regenerate rapidly, they appear to reinnervate skin less successfully than other afferents. Axon regeneration is associated with a recovery of SPLI levels which fell after axotomy: no recovery occurs if regeneration is prevented. Recovery was almost complete in DRG and roots, but incomplete in sciatic nerve. This peptide transmitter in afferent neurons thus behaves in a similar fashion to previously studied low-molecular weight transmitters and related materials in efferent neurons. Since recovery of SPLI levels begins before there is evidence for target reinnervation, it seems that axon regeneration is a sufficient condition for reversal of some axotomy-induced changes in these neurons. Further studies on substance P synthesis and on the response of individual DRG neurons to axotomy and regeneration will be required to explain fully the discrepancy between partial recovery of SPLI levels in sciatic nerve and full recovery in DRG and dorsal roots.     

Regional distribution of kassinin-like immunoreactivity in rat central and peripheral tissues and the effect of capsaicin

The regional distribution of kassinin-like immunoreactivity in rat central and peripheral tissues was investigated by radioimmunoassay and found to resemble closely that of substance P-like immunoreactivity. Neonatal capsaicin treatment caused a similar decrease to both kassinin-like and substance P-like immunoreactivity in primary sensory areas. These results suggest that more than one member of the tachykinin family of neuropeptides exist within the same neuron.     

Three-dimensional distribution of substance P-like immunoreactivity in the urinary bladder of rat

Summary The overall distribution of substance P-like immunoreactive (SPI) fibers in the rat urinary bladder was examined by means of frozen sections and whole mount preparations. Two types of SPI fibers entered the urinary bladder from the neck; one forming thick fiber bundles and the other around the blood vessels. These SPI fiber bundles branched into several thinner segments which often project many collaterals to the smooth muscle and submucosal layers.In the smooth muscle layer, single SPI fibers were seen running parallel to both longitudinal and circular muscle bundles. They appeared to be evenly distributed in the smooth muscle layer.SPI fiber bundles located in the circular muscle layer dissociated SPI fibers which entered the submucosal layer, where they directed to the epithelium and formed a meshwork just beneath it.SPI fibers often left the meshwork to enter the epithelium where abundant arborization of fine SPI fibers was seen. The density of SPI fibers in the submucosal layer and epithelium was not even. The highest density was identified in the neck and the trigonum area.     

Effect of spinal ganglionectomy on substance P-like immunoreactivity in the gastroduodenal tract of cats

Bilateral ligation of the splanchnic nerves and spinal ganglionectomy from T5 to L2 both produced a profound decrease of substance P (SP)-like immunoreactivity in Auerbach's plexuses of the duodenum and the antrum of the stomach in the cats. As unilateral dorsal rhizotomy from T5 to L2 produced no such changes, SP is probably synthesized in the spinal ganglion and transported through the splanchnic nerve to the gastroduodenal wall.     

Substance P-like immunoreactivity in human sympathetic ganglia

The substantia nigra (SN) of the cat was shown, by the anterograde and retrograde horseradish peroxidase methods, to contain neurons which send their axons to the caudomedial portions of the suprageniculate nucleus and/or lateroposterior nucleus of the thalamus; these neurons were located in the restricted region in the lateral part of the SN, which corresponds to the pars lateralis of the SN.     

Immunoreactivity in Limulus: III. Morphological and biochemical studies of FMRFamide-like immunoreactivity and colocalized substance P-like immunoreactivity in the brain and lateral eye

FMRFamide-like immunoreactivity (FLI) and the colocalization of FMRFamide and substance P-like (SPLI) immunoreactivities were examined in the brain and lateral eye of the horseshoe crab with FITC- and TRITC-labeled secondary antibody techniques. In the brain, fibers with FLI were localized in the neuropils of the lamina, medulla, central body, corpus pedunculatum, optic tract, circumesophageal connective, and central neuropil. An extensive network of reactive fibers innervatives the brain's vascular sheath. Somata with FLI were found in the dorsal medial group, dorsal lateral posterior groups #1 and #2, and ventral posterior lateral groups #1 and #2. Several distinct subgroups of reactive somata were noted in both the medullar and ventral medial groups. The distribution of fibers in the brain with colocalized FLI and SPLI includes those which innervate the vascular sheath and widespread populations of small-diameter beaded fibers in the central neuropil and circumesophageal connective. Somata with colocalized FLI and SPLI constitute minority populations in the medullar and dorsal medial groups but form the majority population of a subgroup in the ventral medial group. Overall localization of SPLI was reevaluated and is reported here according to the nomenclature of the new Chamberlain and Wyse brain atlas. In addition to those previously reported, somata with SPLI were found in the dorsal lateral posterior groups #1 and #2, the ventral lateral posterior groups #1 and #2, and several distinct subgroups of the medial and ventral medial groups. In the retina of the lateral eye, fibers with both FLI and SPLI ramify in the lateral plexus and ultimately innervate the corneal epidermis. Brain homogenates were examined for immunoreactive (ir) FMRFamide and ir-substance P with radioimmunoassay techniques. Ir-FMRFamide and ir-substance P eluted in different fractions from both gel filtration chromatography and HPLC. Furthermore, the binding curves for both substances were similar to those of the corresponding synthetic compounds. Brain homogenates were also bioassayed on the lateral eye. Three gel filtration fractions mimic natural circadian activity by increasing the sensitivity of the lateral eye, but they were not coincident with ir-FMRFamide or ir-substance P. Although it is not completely resolved what the active molecules in these fractions are, it is clear that neither ir-FMRFamide nor ir-substance P is a possible candidate.     

The co-occurrence of substance P-like immunoreactivity and dynorphin-like immunoreactivity in striatopallidal and striatonigral projection neurons in birds and reptiles

Using an immunofluorescence procedure that allows the simultaneous labeling of tissue for two different antigens, substance P-like immunoreactivity (SPLI) and dynorphin-like immunoreactivity (DLI) were observed to co-occur extensively in striatal neurons of the avian and reptilian basal ganglia and in fibers and terminals in the projection targets of the avian and reptilian striata. Thus, SPLI and DLI apparently co-occur extensively in striatopallidal and striatonigral projection neurons of the avian and reptilian basal ganglia. Since basal ganglia organization is fundamentally similar among amniotes, the present results suggest that SPLI and DLI may also co-occur extensively in striatal neurons in mammals.     

Evidence for the presence of substance P-like immunoreactivity in the human cerebellum

Preliminary results on the localization of substance P-like immunoreactivity in the human cerebellum are presented. Cerebella from newborn and adult subjects were examined. While only sporadic positive fibres were detected in the adult tissue, the immunoreactive material appeared more abundant in the cerebella from newborn subjects. Varicose and non-varicose fibres and dot-like nerve terminals were present with different density in various regions. The paucity of immunoreactive perikarya suggests that most of the cerebellar substance P-like immunoreactive material has an extrinsic origin.     

Substance P-like immunoreactivity in human prenatal hippocampal formation

Presence and localization of substance P-like immunoreactive neuronal structures in human hippocampal formation during prenatal stages of ontogenesis are reported. In fetuses at 16 and 17 weeks of gestation immunoreactivity is very scarce and represented only by sporadic fibres in Ammon's horn and the entorhinal area. In specimens at 25 and 26 weeks of gestation, more or less intensely labelled perikarya of different morphology are easily detectable in deep layers of Ammon's horn and the hilus of fascia dentata. Immunoreactive beaded fibres are also present at this stage. The possibility of the existence of a substance P-containing extrinsic projection to Ammon's horn is pointed out.     

Distribution of substance P-like immunoreactivity in guinea pig central nervous system

The distribution of substance P-like immunoreactivity (SP-LI) in the guinea pig brain has been studied by immunohistochemistry and the results compared with the distribution in similar regions in the rat brain. In both species, dense SP-LI staining was found in the median eminence, arcuate hypothalamic nucleus, substantia nigra, dorsal raphe and dorsal tegmental nuclei, nucleus of the solitary tract, substantia gelatinosa of the spinal trigeminal nucleus, and spinal cord. Less dense staining was found in the caudate putamen, globus pallidus, nucleus accumbens, habenula, hypothalamic areas, and central grey. SP-LI cell bodies were found in areas previously described for the rat brain including several hypothalamic areas, limbic areas, central grey, and dorsal raphe and solitary tract nuclei. The major difference between the two species was found in the cortex and hippocampus. The guinea pig cortex contained many more SP-LI cells and fibres, distributed in layers II-VI, than the rat cortex. The guinea pig hippocampus contained marked staining, particularly in the pyramidal cell layer of CA1-3 fields of Ammon's horn and in the granular layer of the dentate gyrus, and SP-LI cells in the hilus of the dentate gyrus, whereas rat hippocampus contained few cells and no regions of dense staining. It is concluded that because the guinea pig brain has an extensive distribution of SP-LI in the cortex and hippocampus it resembles the primate brain more closely than does the rat brain.     

Reduced substance P-like immunoreactivity in hereditary sensory neuropathy of Pointer dogs

Summary Two unrelated Pointer dogs, each from a breeding of normal parents which produced three affected pups in a litter of nine, began to bite their paws at 3 and 5 months of age. Insensitivity to painful stimuli was marked in the distal parts of the limbs and receded proximally. The affected dogs were euthanatized at 5 and 20 months because of acral mutilation and infection. Changes affecting the primary sensory neurons included: small spinal ganglia with reduced numbers of cell bodies, degeneration of unmyelinated and myelinated fibers in dorsal roots and peripheral nerves, and reduced fiber density in the dorsolateral fasciculus (dlf).Since nociceptive loss was the salient deficit in a neuropathy affecting primary sensory neurons, immunohistochemical studies focused on substance P, the undecapeptide imputed to mediate nociception at the first synapse in the spinal cord and brain. The localization and density of substance P-like (SPL) immunoreactivity was studied in three control dogs and the two Pointers by the indirect antibody peroxidase-antiperoxidase method. The spinal intumescences of the control dogs contained dense SPL-immuno-reactivity in fibers of the dlf and the superficial laminae of the dorsal horn (i.e., laminae I, II, and the dorsal part of III). Immunoreactive fascicles on the lateral aspect of the dorsal horn and in the reticular process sent contributions medially to a plexiform fiber arrangement in lamina V. Medially, SPL-immunoreactive fibers were more loosely arranged in the internal third of laminae VI and VII and in lamina X. In the Pointers, the loss of primary sensory neurons was associated with notable reduction of SPL staining in the dlf and superficial laminae of the dorsal horn. In the lumbar intumescence of the older Pointer greater preservation of SPL staining in the lateral third of laminae I and II was consistent with somatotopic termination of trunkal afferents in this region.In both Pointers there was no detectable decrease in trigeminal sensitivity and the SPL immunoreactivity in the spinal nucleus of the trigeminal nerve of the younger Pointer and the corresponding control dog appeared equivalent. In the older Pointer, however, the immunoreactivity in this nucleus was decreased relative to the control. This decrease and appearance of scattered fiber degeneration in the dorsal columns of the mature Pointer suggested that fiber degeneration progresses slowly with age to include sensory systems not affected in early postnatal life.The findings in the Pointers were compared with those made in immunohistochemical studies of SP in familial dysautonomia and the mutilated foot rat.Supported in part by a grant from the Rockland County Kennel Club     

Substance P in the human brainstem. Preliminary results of its immunohistochemical localization

The localization of substance P in the human post-mortem brainstem is studied by the indirect immunofluorescence technique. Positive neuronal cell bodies, nerve fibers and terminals are unevenly present over both sensory and effector areas and nuclei. The pattern of distribution of substance P in the human brainstem, similar to that seen in the laboratory animals, supports the hypothesis of a neurotransmitter role of this compound in man.     

Decrease of substance P-like immunoreactivity in the substantia nigra and pallidum of parkinsonian brains

Substance P-like immunoreactivity was measured in the basal ganglia of controls and parkinsonian patients. A significant decrease in substance P-like immunoreactivity was observed within the substantia nigra and the external segment of the globus pallidus from parkinsonian brains. This suggests an alteration of the striatonigral substance P-containing neurons in Parkinson's disease.