P.C. Emson

Distribution of neuropeptide Y-like immunoreactivity in the rat central nervous system—II. Immunohistochemical analysis

The distribution of neuropeptide Y-like immunoreactivity in the rat brain and spinal cord was investigated by means of the peroxidase-antiperoxidase procedure of Sternberger using a rabbit anti-neuropeptide Y serum. A widespread distribution of immunostained cells and fibres was detected with moderate to large numbers of cells in the following regions: olfactory bulb, anterior olfactory nucleus, olfactory tubercle, striatum, nucleus accumbens, all parts of the neocortex and the corpus callosum, septum including the anterior hippocampal rudiment, ventral pallidum, horizontal limb of the diagonal band, amygdaloid complex. Ammons horn, dentate gyrus, subiculum, pre- and parasubiculum, lateral thalamic nucleus (intergeniculate leaflet), bed nucleus of the stria terminalis, medial preoptic area, lateral hypothalamus, mediobasal hypothalamus, supramammillary nucleus, pericentral and external nuclei of the inferior colliculus, interpeduncular nucleus, periaqueductal central gray, locus coeruleus, dorsal tegmental nucleus of Gudden, lateral superior olive, lateral reticular nucleus, medial longitudinal fasciculus, prepositus hypoglossal nucleus, nucleus of the solitary tract and spinal nucleus of the trigeminal nerve. In the spinal cord cells were found in the substantia gelatinosa at all levels, the dorsolateral funiculus and dorsal gray commissure in lumbosacral cord. The pattern of staining was found to be similar to that observed with antisera to avian and bovine pancreatic polypeptide, but to differ in some respects from that observed with antisera to molluscan cardioexcitatory peptide. The presence of neuropeptide Y immunoreactive fibres in tracts such as the corpus callosum, anterior commissure, lateral olfactory tract, fimbria, medial corticohypothalamic tract, medial forebrain bundle, stria terminalis, dorsal periventricular bundle and other periventricular areas, indicated that in addition to the localisation of neuropeptide Y-like peptide(s) in interneurons in the forebrain, neuropeptide Y may be found in long neuronal pathways throughout the brain.

An arcuato-paraventricular and -dorsomedial hypothalamic neuropeptide Y-containing system which lacks noradrenaline in the rat.

The origins of neuropeptide Y-like immunoreactive (NPYI) fibers in the paraventricular and dorsomedial hypothalamic nuclei of the rat were examined using immunohistochemistry. Destruction of the arcuate nucleus resulted in a marked decrease of NPYI fibers ipsilaterally in these nuclei, suggesting that most of NPYI fibers in these nuclei originate from NPYI neurons in the arcuate nucleus. These NPYI systems did not contain noradrenalin.

Distribution of calcitonin gene-related peptide in the rat peripheral nervous system with reference to its coexistence with substance P

This immunocytochemical study, using a double-staining method, showed that calcitonin gene-related peptide-like immunoreactive structures are widely distributed in the peripheral nervous system and that many of them coexist with substance P-like immunoreactive structures in single sensory ganglion cells. Neurons positive for calcitonin gene-related peptide but negative for substance P were detected in sensory ganglia. These cells were large (about 30-45 micron in diameter); these primary sensory neurons containing calcitonin gene-related peptide can probably act independently of substance P. There were neurons containing calcitonin gene-related peptide without substance P in the pterygopalatine ganglion, although these cells were less numerous than in the sensory ganglia. In consecutive sections, calcitonin gene-related peptide-like structures occurred in thyroid parafollicular cells, which also contain calcitonin. This suggested that messenger RNA for producing calcitonin gene-related peptide is also present in the thyroid, and like calcitonin, calcitonin gene-related peptide may have a peripheral physiological role.

Reduced amounts of immunoreactive somatostatin in the temporal cortex in senile dementia of Alzheimer type.

Post-mortem brain tissue from 15 patients dying with a diagnosis of senile dementia of Alzheimer type (SDAT) was compared with tissue obtained from 16 control patients at routine post-mortem. A significant fall in choline acetyltransferase (ChAT) activity was observed in the cortex, hippocampus and amygdala of the SDAT cases and was maximal in the temporal cortex. The fall in ChAT activity observed in the temporal cortex was accompanied by a significant reduction (47%) in immunoreactive somatostatin.

Biochemical and anatomical observations on the degeneration of peptide-containing primary afferent neurons after neonatal capsaicin

Abstract Administration of a single dose of capsaicin (50 mg/kg; sc) to two-day-old rats resulted in a loss of between 85 to 95% of the small diameter unmyelinated primary afferent fibres from lumbar dorsal roots. The numbers of myelinated fibres in the dorsal roots were not significantly affected. The substance P and somatostatin contents of the dorsal roots were decreased by 85 to 95%. The extent of depletion of these peptides from the dorsal horn of the spinal cord was somewhat less; substance P was decreased by 55% and somatostatin by 19%. Histochemical studies showed that fluoride-resistant acid phosphatase was virtually absent from the dorsal horn of capsaicin-treated animals. There was also a conspicuous absence of substance P-positive fibres from layers I and II of the dorsal horn and of fibres traversing these layers to deeper regions. Colchicine pretreatment revealed the presence of substance P-containing perikarya within layers III–IV. The plexus of substance P-positive elements which normally surrounded these neurons was absent after capsaicin treatment, suggesting that they receive a substance P-containing primary afferent innervation. The neurotoxicity of capsaicin after a single dose of 50 mg/kg in neonatal animals appears to be restricted largely to unmyelinated primary afferent fibres. The present results suggest that both substance P and somatostatin are contained in primary afferent sensory neurons which give rise to unmyelinated fibres. It can be further concluded that about 50% of substance P and only about 20% of the somatostatin in the dorsal horn appears to be of primary afferent origin.

Coexistence of calcitonin gene-related peptide and substance P-like peptide in single cells of the trigeminal ganglion of the rat: immunohistochemical analysis

The localization of calcitonin gene-related peptide (CGRP) and substance P (SP) in the rat trigeminal ganglion was examined by means of the indirect immunofluorescent method. About 40% of neurons in the ganglion contained CGRP-like immunoreactivity (CGRPI), while about 20% of neurons showed SP-like immunoreactivity (SPI). In serial sections, nearly all the SPI neurons contained CGRPI.

Topographic localization of calcitonin gene-related peptide in the rat brain: An immunohistochemical analysis

Abstract The distribution of immunoreactive calcitonin gene-related peptide in the rat brain was investigated by means of an indirect immunofluorescence method. In addition to previously reported calcitonin gene-related peptide-like immunoreactive structure-containing sites such as the nucleus ambiguus, nucleus originis nervi facialis, nucleus originis nervi hypoglossi, nucleus peripeduncularis and nucleus parabrachialis, the present study demonstrated a far wider distribution of calcitonin gene-related peptide-like immunoreactive structure-containing cells in the rat brain, i.e. the nucleus hypothalamicus lateralis. nucleus ventromedialis thalami, colliculus superior, lemniscus lateralis, gyrus dentatus. nucleus olivaris superior, nucleus tractus solitarii, nucleus cuneiformis, nucleus parabigeminalis and a proportion of the Purkinje cells. We have also demonstrated a more extensive network of calcitonin gene-related peptide-like immunoreactive fibers distributed in various areas throughout the rat brain than has been reported previously such as the colliculus inferior, nucleus olivaris superior, nucleus vestibularis lateralis and inferions, and nucleus cochlearis dorsalis and ventralis, etc.

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 Huntingtons 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 Huntingtons 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 Huntingtons disease.

Nerve growth factor counteracts the neurophysiological and neurochemical effects of chronic sciatic nerve section

The sciatic nerve was sectioned unilaterally in rats and nerve growth factor (NGF) applied locally to the nerve stump for the following 10-14 days using an indwelling osmotic pump. The aim of the experiment was to test whether NGF had any effect on the previously reported neurophysiological and neurochemical events that occur central to a peripheral nerve lesion. The method of application allowed the sciatic nerve on the other side to be used as a control. Primary afferent depolarization fell, as expected, to 13% of its control value after chronic nerve section but if NGF was administered it fell to only 43.5% of control. Chronic nerve section is also known to result in expansion of the receptive fields of deafferented dorsal horn cells. NGF treatment reduced the number of such large receptive fields by 50%. The normal depletion of fluoride resistant acid phosphatase from the cut nerve terminals in the dorsal horn did not occur following NGF treatment. Radioimmunoassay of substance P revealed that the 30% reduction in dorsal horn levels that follows chronic sciatic nerve section did not occur when NGF was applied and that the accompanying 60% decrease in dorsal root ganglion levels was changed to a 64% increase by NGF. The results show that chronic NGF treatment of a cut sciatic nerve does partially reverse the central changes that normally follow deafferentation.

Survival of basal ganglia neuropeptide Y-somatostatin neurones in Huntington's disease.

The content of somatostatin-like immunoreactivity (SRIF-LI) and neuropeptide Y-like immunoreactivity (NPY-LI) has been measured in both control post-mortem human brains and in Huntingtons disease brains. The content of both SRIF-LI and NPY-LI was found to be significantly increased in the basal ganglia of Huntingtons disease brains compared with a control group. The nature of the SRIF-LI and NPY-LI in both control and Huntingtons disease brains was investigated after separation on Sephadex G25 and G50 columns. Using a C-terminal-directed SRIF radioimmunoassay (RIA), 3 peaks of immunoreactivity were measured, whilst an N-terminal-directed SRIF RIA detected two peaks of immunoreactivity. In each case, the elution profile did not differ between control and Huntingtons disease caudate nucleus. The content of immunoreactivity in each peak was found to be increased in Huntingtons disease brains compared with controls. Only one peak of NPY-LI was detected in both control and Huntingtons disease caudate after separation on Sephadex G25 and G50 columns. Immunohistochemical staining of the caudate and putamen of control and Huntingtons disease brains revealed a population of neurones containing NPY-LI. The number of NPY-positive neurones was found to be increased in both the caudate and putamen of Huntingtons disease brains compared to control caudate and putamen.