Prenatal ontogeny of substance P-like immunoreactivity in the parabrachial nucleus of the human fetus — an immunocytochemical study

Using an immunocytochemical method, the prenatal ontogeny of substance P-like immunoreactivity (SP-LI) was demonstrated in the parabrachial nucleus (PB) of human fetus, fetal age (menstruation age) 11.5 to 40 weeks. The time of initial appearance of SP-LI in the human brainstem PB was between fetal age 11.5 weeks and 16 weeks. While the fetus grew, the density of SP-LI fibers and terminals in the human fetus brainstem PB increased constantly from fetal age 16 weeks to 40 weeks. These findings indicate that substance P (SP) might play an important role in the human parabrachial nucleus development and in its functional establishment during the prenatal period.     

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.     

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.     

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     

Prenatal ontogeny of substance P-like immunoreactivity in the trigeminal spinal caudal subnucleus of the human fetus — an immunocytochemical study

Using an immunocytochemical method, the prenatal ontogeny of substance P-like immunoreactivity (SP-LI) was demonstrated in the trigeminal spinal caudal subnucleus (TSCS) of human fetus, fetal age (menstruation age) 11.5–40 weeks. The time of initial appearance of SP-LI in the human brainstem TSCS was between the fetal age 11.5 weeks and 16 weeks. While the fetus grew, the density of SPLI fibers and terminals in the human fetus brainstem TSCS increased constantly from fetal age 16 weeks to 40 weeks. These findings indicated that substance P (SP) might play an important role in the human trigeminal nerve system development and in its functional establishment during the prenatal period.     

Distribution of substance P-like Immunoreactivity in the chameleon brain

The distribution of substance P-like immunoreactivity in the chameleon brain and spinal cord was studied with immunohistochemical methods using polyclonal antibodies against substance P. In the telencephalon, immunoreactive cell bodies and fibers were located primarily in the striatum and in the globus pallidus. In addition, few substance P-like fibers were observed in the cortical areas, in the septum, and in the amygdala. In the diencephalon, a high density of immunostained neurons and fibers were seen in the periventricular and ventrolateral hypothalamus. Another group of cell bodies was located in the optic tectum and particularly in the stratum griseum central. A large number of immunoreactive fibers were also detected in the thalamic nuclei and in the median eminence. In the mesencephalon, few immunoreactive neurons were observed in the ventral tegmental area, in the substantia nigra, and in the nucleus reticularis isthmi. These latter nuclei, the periventricular area, the posterior commissure, the nucleus lentiformis mesencephali, the oculomotor nucleus, and the raphe nuclei contained a dense plexus of substance P immunoreactive fibers. No immunoreactive cell bodies were observed in raphe nuclei. In the spinal cord, no substance P-like immunoreactive neurons were observed, but a large number of substance P immunostained fibers were seen in the dorsal and lateral part of the dorsal horn and surrounding the dorsal parts of the central canal. The results of the present study are discussed with respect to those obtained in other species of reptiles, the main differences concerning the lateral septum, the habenula, the area of the paraventricular organ, and the raphe nuclei.     

Prenatal ontogeny of substance P-like immunoreactivity in the nucleus tractus solitarii and dorsal motor nucleus of the vagus nerve of the human fetus: an immunocytochemical study

By using immunocytochemical method, the prenatal ontogeny of substance P-like immunoreactivity (SP-LI) was demonstrated in the dorsal motor nucleus of the vagus nerve (nX) and the nucleus tractus solitarii (nTS) of the human fetus at fetal age (menstruation age) of 11.5 weeks to 40 weeks. The time of initial appearance of SP-LI in the human brainstem nTS was between the fetal age 11.5 weeks and 16 weeks. At fetal age 16 weeks, the nTS showed moderate density of SP-LI fibers and terminals in subnucleus dorsalis of the nTS and nX. While the fetus grew, the density of SP-LI in the human fetus brainstem nTS and nX increased gradually from fetal age 16 weeks to 40 weeks. According to the Nissl staining, at fetal age 23 weeks, the nTS of human fetus can be subdivided into dorsal, medial, dorsolateral, ventrolateral and ventral gelatinosus subnuclei. The cytoarchitectonic subdivisions of human fetus nTS is in good agreement with the results obtained by immunocytochemical staining. These findings indicated that substance P (SP) might play an important role in the development of human brainstem nX, nTS, their related cranial nerves, and in their functional establishment during the pranatal period.     

Quinolinic and kainic acids can enhance calcitonin gene-related peptide-like immunoreactivity in striatal neurons with substance P-like immunoreactivity

Quinolinic or kainic acid, when injected into the cat striatum, caused neurodegenerative changes in striatal neurons. In parallel with these changes, however, calcitonin gene-related peptide-like immunoreactivity became visible in perikarya of many striatal neurons which were distributed around the lesions produced by the quinolinic or kainic acid injections. The vast majority of these lesion-spared neurons showing calcitonin gene-related peptide-like immunoreactivity were striatal neurons with substance P-like immunoreactivity.     

Comparison of substance K-like and substance P-like fibers and cells in the rat hippocampus

Substance P (SP) and substance K (SK) are structurally related peptides which are both encoded in the preprotachykinin A gene. The distribution of SP- and SK-like fibers and cell bodies in the rat hippocampus were studied by immunohistochemistry. The distribution of SK-like fibers was similar to that of SP-like fibers but there were few SK-like fibers. Fibers for both peptides were prominent in the dorsal and ventral subiculum and at the junction of CA₂ and CA₃. SP- and SK-like cell bodies were noted in the subiculum and in the stratum oriens of CA₁ and CA₂. SP- and SK-like cells were also noted in the ventral dentate gyrus but only SP-like cells were found in the dorsal dentate gyrus.     

Immunohistochemical demonstration of coexistence of enkephalin- and substance P-like immunoreactivities in axonal components in the lumbar segments of cat spinal cord

Coexistence of enkephalin-like and substance P-like immunoreactivities (ENK-LI and SP-LI) was shown in axonal components in the lumbar segments of cat spinal cord by using the double immunofluorescence method. Although axonal components labeled with both ENK-LI and SP-LI were distributed throughout the spinal gray, they were most dense in the ventral horn, and most sparse in the superficial part of the dorsal horn.     

Distribution of substance P-like immunoreactive neurons in the human medulla oblongata: co-localization with monoamine-synthesizing neurons

The raphe nuclei also contained SP-like immunoreactivity (up to 30%) while few monoamine-synthesizing neurons in the lateral and dorsomedial medulla contained SP-like immunoreactivity (approximately 5% of presumed serotonin-, noradrenaline-, and adren- the adult human. The majority of SP-like immunoreactive neurons were found in four main regions: the lateral medulla, the dorsomedial medulla, the spinal trigeminal nucleus, and the raphe nuclei. The morphology of immunoreactive cells varied according to the region in which they were found. In contrast to previous studies, we found large numbers (90,000) of SP-like immunoreactive neurons throughout the adult human medulla oblongata. The distribution of these SP-like immunoreactive neurons appears to be significantly different from those described in the rat and cat. These results were compared to the distributions of monoamine-synthesizing and neuropeptide Y (NPY)-like immunoreactive neurons in the human medulla previously reported (Halliday et al.: Neuroscience, in press, 1988a; J. Comp. Neurol., in press, 1988b). Colocalization studies revealed that many presumed serotonin-synthesizing neurons in the raphe nuclei also contained SP-like immunoreactivity (up to 30%) while few monoamine-synthesizing neurons in the lateral and dorsomedial medulla contained SP-like immunoreactivity (approximately 5% of presumed serotonin-, noradrenaline-, and adrenaline-synthesizing neurons). The distributions of SP- and NPY-like immunoreactive neurons were similar, although SP-like immunoreactive neurons were concentrated in the lateral regions of the same structures. We have found that the distributions of monoamine-synthesizing, NPY-, and SP-like immunoreactive neurons significantly overlap, particularly in the lateral medulla of the adult human. There is a large increase in the number of these cells in this region compared to other species, emphasizing the neuroanatomical differences between humans and other species.     

Substance P-like immunoreactivity and somatostatin-like immunoreactivity in the ventricular fluid of patients with chronic pain syndromes

Summary Substance P-like and somatostatin-like immunoreactivities (SPLI and SLI) were determined in ventricular fluid of patients with chronic pain syndromes and in a comparison group with multiple sclerosis, essential tremor, epilepsy and postanoxic myoclonus. Concentrations of SPLI and SLI were non-significantly decreased by 40% and 33% in chronic pain patients as compared with control patients without pain. There were no differences apparent between subgroups of pain patients (deaferentation pain, neoplasia-induced pain, thalamic pain). High pressure liquid chromatography combined with radioimmunoassay showed marked heterogeneity of SPLI and SLI.     

Characterization and partial purification of a 5,700 dalton form of substance P-like immunoreactivity from the rat hypothalamus

Analysis of rat hypothalamic extracts by gel filtration and HPLC revealed the presence of a form of substance P-like immunoreactivity with a molecular weight of approximately 5,700 (6k-SP). 6k-SP is probably an N-terminally extended form of substance P; the peptide may be an intermediate in SP biosynthesis, or may function as a biologically active peptide in its own right.     

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 substance P immunoreactive nerves in the carotid body

Immunofluorescent histochemical observation of the rat carotid body revealed the presence of substance P immunoreactive fibers. This supports the hypothesis that substance P is a neurotransmitter of chemoreceptor afferent nerves.     

Regional distribution of substance P-like immunoreactivity in the lower brainstem of the rat

A radioimmunoassay was used to quantify substance P-like immunoreactivity in the lower brainstem of the rat. The peptide was detectable in all brainstem nuclei investigated. A very high concentration was found in the substantia nigra, and high concentrations in the interpeduncular nucleus and central gray matter. Levels in the pons, the parabrachial nuclei; in the medulla, the nucleus tractus spinalis of the Vth nerve and the nucleus of the solitary tract were higher than the brain average.     

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.     

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.     

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.     

Anatomical distribution of substance P-immunoreactive neurons in human brainstem during the first postnatal year

The localization of substance P (SP)-immunoreactive structures in the infant brainstem was investigated by immunohistochemistry using the peroxidase antiperoxidase technique. SP-immunoreactive structures are widely distributed throughout the brainstem region. SP-immunoreactive cell bodies are prominent in the superior colliculis, the central grey, the nucleus tractus solitarius and the reticular formation. A high density of SP-immunoreactive fibers is found in the nucleus tractus solitarius, the trigeminal nucleus and the dorsal horn of the spinal cord. Large SP-immunoreactive fibers are seen in the substantia nigra. In the present study, we also investigated the development of substance P-immunoreactive fibers in the infant brainstem during the first postnatal year. We note a qualitative increase in the density of SP-immunoreactivity in some brainstem regions such as colliculus superior and substantia nigra with respect to age.