Substance P-like immunoreactivity in human cerebrospinal fluid

In a study to elucidate molecular mechanisms in pain, substance P-like immunoreactivity (SP-LI) was measured in lumbar CSF from 75 patients with lower back pain. Two samples--one before and one after lidocaine treatment--were obtained from each patient, and total SP-LI was measured in unfractionated (no HPLC) samples. SP-LI data were separated into three categories: placebo responders, pharmacological responders, and pharmacological non-responders. A significant difference was observed between the total SP-LI measurement of first and second samples of pharmacological non-responders. Distribution of SP-LI immunoreactive molecular species in two CSF patient samples (no ODS) was analyzed with a combination of reversed phase (RP) HPLC and RIA. Immunoreactivity in collected HPLC fractions was measured at calibrated retention times of synthetic SP-sulfoxide (SP-O), SP, and SP. Qualitative and quantitative differences in those HPLC-RIA metabolic profiles were observed within and between those two patients' samples. These data indicate that the type and amount of SP metabolism and SP precursor-processing differs in CSF between these two representative patients and within the short amount of time elapsed between acquiring these two samples.     

Chemical and immunochemical characterisation of substance P-like immunoreactivity and physalaemin-like immunoreactivity in a carcinoid tumour

Extracts of a carcinoid tumour, resected from the mid-portion of the ileum of a patient with no symptoms of endocrine disorder, were associated with a high concentration of substance P-like immunoreactivity. Using reverse-phase high performance liquid chromatography and antisera specifically directed against the C-terminal and N-terminal regions of substance P and against the N-terminal region of physalaemin, the following components were isolated and identified: substance P and its oxidised form, [pGlu⁵]substance P-(5–11) peptide and its oxidised form, and the oxidised form of physalaemin. The identity of tumour substance P with the undecapeptide was confirmed by amino acid analysis and high performance ion-exchange chromatography. In vitro incubation of tumour tissue from a lymph node metastasis from the same patient with [³H]leucine resulted in incorporation of radioactivity into newly synthesised substance P.     

Enteroglucagon and substance P-like immunoreactivity in argentaffin and argyrophil rectal carcinoids.

A specific immunofluorescence for enteroglucagon or substance P or for both hormones was demonstrated in nine out of 12 examined rectal carcinoids. One tumor was argentaffin, contained ultrastructurally pleomorphic granules of the entero-chromaffin cell type, and showed immunofluorescence for substance P. The rest were non-argentaffin but were argyrophil with the Grimelius technique and contained round granules. The argyrophil carcinoids were immunoreactive to one or both hormones in eight cases and not fluorescent in three cases. In two of the non-argentaffin carcinoids a small number of argyrophil cells was found with the method of Sevier-Munger.     

Neurokinin A-like immunoreactivity in feline dental pulp: its distribution,origin and coexistence with substance P-like immunoreactivity

Summary The distribution and origin of neurokinin A (NKA)-like immunoreactivity were investigated in feline dental pulp by an indirect immunofluorescence method. NKA-containing nerve fibres with varicosities, which entered the dental pulp via apical foramen, were distributed throughout this tissue. Many NKA-containing nerve fibres were localized around blood vessels, but some were observed apart therefrom. At the odontoblastic layer, thin NKA-containing nerve fibres were observed running straight toward the pulp-predentinal border between odontoblasts. After inferior alveolar nerve section, all NKA-containing nerve fibres disappeared in the dental pulp, while the removal of the superior cervial ganglion resulted in no change in the distribution of these fibres. The correlation of NKA-like immunoreactivity and substance P (SP)-like immunoreactivity was also investigated by double-immunofluorescence technique. The distribution of NKA-containing nerve fibres was very similar to that of SP-containing nerve fibres; it appeared that all NKA-containing nerve fibres contained SP.     

Distribution of substance P-like immunoreactivity in the pedal ganglion of Mytilus galloprovincialis

Nerve cell bodies and fibers displaying Substance P-like (SP-li) immunoreactivity have been identified in the pedal ganglia (PG) of Mytilus galloprovincialis. Most reactive neurons are small-sized, unipolar and are located in the ganglionic cortex. Positive fibers form a dense network in the neuropile and run parallel in the commissure and in the nerves and connective tracts. The positivity of the labelled elements to a monoclonal antibody indicates that SP-li substance of Mytilus contains a portion immunologically similar to the C-terminal pentapeptide responsible for the pharmacological activity of vertebrate SP.     

Effect of cisapride on the concentrations of beta-endorphinlike immunoreactivity and substance P-like immunoreactivity in the rat gastrointestinal tract

Cisapride is a gastrointestinal prokinetic agent reported to be devoid of direct cholinergic effect from the myenteric plexus of the gut. The effect of cisapride (0.125, 0.5, 2mg/kg, i.p.) on the concentration beta-endorphin and substance P in rat gastrointestinal tract was studied. beta-Endorphinlike immunoreactivity contents were significantly increased in both mucosal and muscular layers of the entire gastrointestinal tract (from gastric body to rectum) of the rats treated with 2 mg/kg of cisapride. beta-Endorphinlike immunoreactivity contents were also increased in a part of the gastrointestinal tract of the rats treated with 0.125 or 0.5 mg/kg of cisapride. Substance P like immunoreactivity contents were significantly decreased in muscular layers of the rectosigmoid colon of the rats treated with 2 mg/kg of cisapride. This study suggests that the prokinetic effects of cisapride may relate to the contents of beta-endorphinlike immunoreactivity and substance P like immunoreactivity in gastrointestinal tract.     

Localization of substance P-like immunoreactivity in the palate and trigeminal ganglion of Rana pipiens

This study was undertaken to localize substance P-like immunoreactivity (SP) in the nerve fibers innervating the palate, identify the ganglion of the palatine nerve and determine whether it contains SP cell bodies, in the frog Rana pipiens. The palatine nerve which is a branch of the maxillo-mandibular subdivision of the trigeminal nerve was traced to the trigeminal ganglion that connects to the medulla by the trigeminal nerve root. Using an immunocytochemical method, SP containing fibers with varicosities were found in the connective tissue layer of the palate. Some of these fibers were observed adjacent to blood vessels to the epithelial layer of the palate in apparent innervation of the ciliated epithelial and mucus cells. SP-labeling was also observed in small to medium cells of the trigeminal ganglion. These results appear to support the pharmacological studies of SP on the regulation of mucociliary activity in the frog R. pipiens.     

Neurokinin B- and substance P-like immunoreactivity are co-localized in enteric nerves of rat ileum

The tachykinins (TKs) substance P (SP), neurokinin A (NKA), and neurokinin B (NKB) have conserved C-terminal sequences and mediate similar physiological responses by activating neurokinin receptors found on neural and smooth muscle cells. Many enteric nerves express preprotachykinin A (PPT A) mRNA and synthesize SP and NKA. However, it is unclear if NKB is synthesized in enteric neurons as many antibodies developed against NKB also recognize other TKs. Therefore, the cellular distribution of NKB-like-immunoreactivity (NKB-ir) in rat ileum was examined using selective antisera raised against either synthetic Cys10-NKB or peptide 2 (P2), a non-tachykinergic peptide sequence in NKB precursor protein. NKB-ir and P2-ir had a similar distribution in varicose nerve fibers in submucosal and myenteric ganglia and almost all ganglia contained immunoreactive nerves. Few submucosal or myenteric neuronal somata contained strong immunoreactivity. Preabsorption of NKB or P2 antisera with their respective cognate peptides, but not with other TK peptides, abolished specific immunostaining. Finally, co-localization of NKB-/P2-ir with SP-ir suggested that most NKB-/P2-ir nerve fibers contain SP-ir, but some SP-ir nerves do not contain detectable NKB-/P2-ir. These results indicate that PPT B products P2 and NKB are localized in a subpopulation of enteric nerves containing TKs encoded by PPT A. Stimulation of these nerves may release NKB to activate local neurokinin receptors.     

Methionine enkephalin-like immunoreactivity, substance P-like immunoreactivity and β-endorphin-like immunoreactivity postmortem stability in rat pituitary

Significant post-mortem changes in peptide concentration occur within the previously unstudied timeframe, i.e. within 1 h, for the proenkephalin A, proopiomelanoccortin, and tachykinin neuropeptidergic systems in the pituitary. These data differ from data obtained in other studies that concluded that peptides are stable for up to 72 h post-mortem. The post-mortem stability of the three neuropeptides, methionine enkephalin, substance P, and β-endorphin, was studied in the rat pituitary to test the hypothesis that significant post-mortem concentration changes of those three neuropeptides occur in the immediate post-mortem time period.     

Striatal substance P-like immunoreactivity: characterization of high-performance liquid chromatography and aspects of subcellular distribution and in vitro release by potassium

Substance P-like immunoreactivity (SPLI) in acetic acid extracts of striatum elutes in a single peak that corresponds to that for synthetic substance P, using reverse-phase high-performance liquid chromatography. This finding suggests that striatal SPLI, as detected with the radioimmunoassay used here, largely represents a single molecular species. Striatal SPLI is highly concentrated in the crude mitochondrial fraction (P2); over 70% of this P2-SPLI is recovered in the synaptosomal subfraction. The potassium-induced release of SPLI from striatal tissue in vitro occurs only when enough extracellular calcium is available. These data suggest that substance P in striatal neurons may participate in neural transmission locally, exclusive of its probable role in maintaining the substance P supply to nerve terminals in the substantia nigra.     

Light and electron microscopic localization of substance P-like immunoreactivity in the cerebral ganglion of locust with a monoclonal antibody

Summary The occurrence of substance P-like immunoreactivity was studied in the locust brain at light and electron microscopic level using monoclonal IgG fraction to substance P. Small immunoreactive perikarya have been found beside the medial neurosecretory cells in horizontal brain sections. Widespread immunoreactivity was also observed in the protocerebral neuropil notably in the central body and bordering on the corpora pedunculata. The reaction endproduct appeared as fine, more or less round particles in the central body, and as coarse varicosities and wavy fibres bordering the peduncles. The roundish particles probably represent nerve terminals, while the wavy fibers correspond to neural processes. In the vicinity of the lobe immunoreactivity was not observed. Electron microscopically, a number of immunoreactive terminals were found in the protocerebral neuropil. The reaction endproduct was accumulated mostly in large dense core granules/average diameter 80 nm/however reaction endproduct was also observed on the external surface membranes of clear vesicles and mitochondria. Our results suggest the widespread occurrence of a substance-P immunoreactive neuropeptide in the cerebral ganglia of the migratory locust.     

Characterization of substance P-like immunoreactivity in peripheral sensory nerves and enteric nerves by high pressure liquid chromatography and radioimmunoassay

Material exhibiting immunoreactivity for substance P in enteric nerves, obtained from the myenteric plexus of the guinea pig small intestine, and in the peripheral ends of sensory nerves of the ureter, atrium and superior mesenteric artery, was characterized by separation by high pressure liquid chromatography, and quantified by radioimmunoassay of fractions collected from the chromatograph. Capsaicin, which depletes substance P-like immunoreactivity from sensory, but not from other substance P-containing nerves, reduced the content of substance P-like immunoreactivity in ureter, atrium and superior mesenteric artery by more than 99.5%, whereas the reduction in immunoreactive material in the myenteric plexus was less than 10%. Separation of extracts of myenteric plexus, ureter and atrium on a reversed-phase column gave major peaks corresponding to authentic substance P and minor peaks that coeluted with oxidized substance P. If the extracts were oxidized with hydrogen peroxide before chromatography, all the immunoreactivity was found in the peak corresponding to oxidized substance P. In the superior mesenteric artery extracts, in addition to the components corresponding to substance P and its oxidized derivative, there was a small intermediate peak that has yet to be identified. Physalaemin, which has been suggested to be present in mammalian nerves, was not detectable in any of the extracts. It is concluded that both enteric nerves and the peripheral processes of sensory nerves which show immunoreactivity for substance P in this species contain the authentic peptide.     

Transient appearance of substance P-like immunoreactivity in the granular convoluted tubules of the male mouse submandibular gland: light- and electron-microscopic studies

The time of appearance and distribution of substance P (SP)-like immunoreactivity in the granular convoluted tubule cells of the developing male mouse submandibular glands were examined, and the subcellular localization of SP-like immunoreactivity was investiagted by electron microscopy. At 25 days of age, SP-like immunoreactivity was first detected in the supranuclear cytoplasm of the granular convoluted tubule cells, which occurred either singly or in small clusters. At 30 and 35 days of age, granular convoluted tubule cells with SP-like immunoreactivity were more numerous than in the earlier stages, as the volume ratio of the cells increased. Not all granular convoluted tubule cells demonstrated SP-like immunoreactivity. The number of cells with SP-like immunoreactivity decreased at 60 days of age, and these cells had completely disappeared at 90 days of age. Most, but not all, secretory granules in the granular convoluted tubule cells were strongly labeled with gold particles, indicating that the subcellular site of SP-like substance is in the secretory granules within the cells. The findings suggest that the physiological role of the SP-like substance secreted from the GCT cells is restricted to the early postnatal stages, and that it may be involved in the development of the oral mucosa or digestive tract as a trophic factor.     

Calcitonin gene-related peptide-like immunoreactivity in nerve fibers in the human skin. Relation to fibers containing substance P-, somatostatin- and vasocactive intestinalpolypeptide-like immunoreactivity

Calcitonin gene-related peptide-like immunoreactivity was demonstrated in in sensory nerve fibers in the epidermis and dermis as free nerve endings and around blood vessels and hair follicles of the human finger pad and arm skin. The vast majority of the calcitonin gene-related immunoreactive fibers was shown to display also substance P-like immunoreactivity and a few fibers in the dermis were somatostatin positive. No fibers displaying both substance P and somatostatin-like immunoreactivity were found but a few substance P immunoreactive fibers in the dermis-epidermis region were found to contain also vasointestinal polypeptide-like immunoreactivity. In the sweat glands, abundant calcitonin gene-related peptide positive, but substance P negative, fibers were observed with a similar distribution pattern as the vasoactive intestinal polypeptide immunoreactive fibers and these fibers were suggested to be of sympathetic origin.     

Systemically administered capsazepine prevents the capsaicin-induced functional desensitization and loss of substance P-like immunoreactivity (SP-LI) in guinea-pig bronchi

In this study, we investigated whether the systemically administered capsazepine can prevent the capsaicin-induced desensitization ex vivo in guinea-pig bronchi. Pretreatment with capsaicin (2.5, 5 and 10 mg/kg, s.c.) induced the functional desensitization and the loss of substance P-like immunoreactivity (SP-LI) with a similar potency (ED50: 3.31 +/- 0.57 and 4.81 +/- 0.89 mg/kg, respectively) in isolated guinea-pig bronchi. Capsazepine (30 mg/kg, s.c.) co-administered with capsaicin (5 mg/kg, s.c.) prevented the capsaicin (5 mg/kg, s.c.)-induced functional desensitization and loss of SP-LI. These results suggest that capsazepine can antagonize systemically the desensitizing action of capsaicin at the level of receptor, preventing the loss of SP-LI and the establishment of functional desensitization in guinea-pig bronchi.     

Distribution of serotonin-, Gamma-Aminobutyric Acid- and substance P-like immunoreactivity in the central and peripheral nervous system of Mytilus galloprovincialis

The distribution of serotonin-, GABA- and substance P-like immunoreactivity has been studied in the cerebral and visceral ganglia and in some peripheral tissues of Mytilus galloprovincialis (Moleusca, Bivalvia). Cerebral ganglia contain a developed serotonin-immunoreactive neuronal subpopulation and numerous GABA-immunoreactive neurons, whereas neurons positive for substance P are sparse. In peripheral tissues innervated by the cerebral ganglia (labial palps and oesophagus) only serotonin-immunoreactive nerve fibers were found. In the visceral ganglia, serotonin- and GABA-immunoreactive neurons are far less numerous than in the cerebral ganglia, whereas several neurons positive for substance P are scattered in all cortical zones. Serotonin-immunoreactive plexuses innervate the posterior adductor muscle and the gill filaments which contain also a developed nerve network positive for substance P. The distribution pattern of the immunoreactive elements in the ganglia and in peripheral territories indicates that GABA should exert only a central action, whereas serotonin and a substance P-like peptide are involved both in central and peripheral neurotransmission.