Coexistence Of Calcitonin Gene-related Peptide Research Articles

Distribution and origin of secretoneurin-immunoreactive nerves in the female rat uterus

Secretoneurin is a 33-amino acid peptide derived from secretogranin II. Secretoneurin immunoreactivity has been localized in the peripheral nervous system where it exerts potent chemotactic activity for monocytes and may play a role in inflammation. Secretoneurin could play a role in this process, although the presence and distribution of secretoneurin-immunoreactive neurons in the female reproductive system has not been documented. Thus, this study was undertaken to examine secretoneurin immunoreactivity in nerves of the rat uterus and uterine cervix. A moderate plexus of secretoneurin-immunoreactive nerve fibers was present in the myometrium and endometrium of the uterus as well as in the smooth muscle and endocervix of the cervix. Many of these fibers were associated with the vasculature as well as the myometrium. Secretoneurin immunoreactivity was present in small- to medium-sized neurons of dorsal root and nodose ganglia. Retrograde tracing with FluoroGold indicated that some of these sensory neurons project axons to the cervix and uterine horns. Secretoneurin-immunoreactive terminal-like structures were associated with neurons in the sacral parasympathetic nucleus of the lumbosacral spinal cord. In addition, some secretoneurin terminals were apposed to pelvic parasympathetic neurons in the paracervical ganglia that projected axons to the uterus and cervix. Double-immunostaining indicated co-existence of calcitonin gene-related peptide or substance P with secretoneurin in some sensory neurons, in some terminals of the pelvic ganglia, as well as nerve fibers in the uterine horn and cervix. Finally, fibers in the uterus and cervix were depleted of secretoneurin by capsaicin treatment. This study indicates that secretoneurin is present in the uterus in C-afferent nerve fibers whose cell bodies are located in sensory ganglia. Some of these fibers contain both secretoneurin and calcitonin gene-related peptide or substance P. These substances have functions in inflammatory reactions. Further, secretoneurin could influence postganglionic parasympathetic “uterine-related” neurons in the pelvic ganglia and preganglionic parasympathetic neurons in the lumbosacral spinal cord.

The coexistence of calcitonin gene-related peptide and substance P in pericellular arborization and satellite cell of goat trigeminal and nodose ganglia.

Pericellular arborization is reported to be the self-regulating structure in sensory ganglia. Although the calcitonin gene-related peptide (CGRP) or substance P (SP) immunoreactive pericellular arborization appeared in the sensory ganglia, there was no available information that CGRP and SP colocalize in this structure. As the attempts to resolve the question described above, the present study was undertaken to identify the coexistence of CGRP and SP in pericellular arborizations of the goat nodose and trigeminal ganglia by double immunohistochemistry. As the results show, CGRP immunoreactivity was present in every pericellular arborization containing SP immunoreactivity in trigeminal ganglia, however, pericellular network containing CGRP or SP immunoreactivity was not present in nodose ganglia. Unexpectedly, a few small satellite elements were observed to contain intense CGRP and SP immunoreactivity at the periphery of CGRP and SP immunoreactive neurones in nodose ganglia. Therefore, these results suggest that CGRP and SP coexist in pericellular arborizations, and that satellite cell as well as pericellular arborization may be involved in intraganglionic regulation of goat sensory ganglia.

Coexistence of calcitonin gene-related peptide and NADPH-diaphorase in the canine superior cervical ganglion

By means of double staining technique of NADPH-diaphorase (NADPH-d) histochemistry and calcitonin gene-related peptide (CGRP) immunohistochemistry, we investigated the coexistence of NADPH-d reactivity and CGRP immunoreactivity in the canine superior cervical ganglion (SCG). Most of NADPH-d reactivity and CGRP immunoreactivity were coexisted in the principal postganglionic neurons. These neurons were distributed throughout the ganglion without specific localization. The present findings suggest the intimate role of CGRP and nitric oxide in postganglionic neurons of the canine SCG.

The cellular localization of the neuropeptides substance P, neurokinin A, calcitonin gene-related peotide and neuropeptide Y in guinea-pig vestibular sensory organs: a high-resolution confocal microscopy study

Four neuropeptides, substance P, neurokinin A, calcitonin gene-related peptide and neuropeptide Y, were detected by radioimmunoassay in guinea-pig vestibular end-organs. High-resolution confocal microscopy visualization of immunofluorescence staining was used to determine the cellular localization of these peptides. Substance P- and neurokinin A-like immunoreactivities were found to co-exist in afferent fibers innervating the peripheral regions of both the utricular and ampullar sensory organs. The immunoreactivity was more concentrated in the distal ends of the calyceal-shaped nerve endings that innervate type I sensory cells. While in the guinea-pig, nerve calyces and type I cells are distributed in both the central and peripheral regions of the sensory epithelia, immunoreactive calyces were found only in the peripheral regions. Calcitonin gene-related peptide-like immunoreactivity was localized in small bouton endings situated at the level of the base of the hair cells. These boutons were in a position to make axosomatic contacts with type II sensory cells and axodendritic contacts with afferent nerve endings. Calcitonin gene-related peptide immunoreactivity co-existed with choline acetyltransferase immunoreactivity. The localization and shape of these boutons identified them as the axonal endings of efferent vestibular fibers. Neuropeptide Y-like immunoreactivity was not observed in the actual sensory epithelium but in the underlying connective tissue, where it was located in varicose fibers along blood vessels.The synaptic position of the tachykinins is clearly distinct from that of calcitonin gene-related peptide. This segregation distinguishes the vestibular end-organs from most peripheral tissues where these peptides are co-localized. The tachykinin-immunoreactive afferent fibers are postsynaptic to the hair cells. If, as in somatic sensory endings, these fibers can be triggered to release the neuropeptides by an axon reflex type of activation, then the tachykinins could interfere directly with the function of type I and type II vestibular hair cells. Calcitonin gene-related peptide co-exists with acetylcholine in the efferent axonal endings that are presynaptic to type II hair cells and to afferent fibers. Calcitonin gene-related peptide can thus interfere by direct synaptic action with type II hair cells only. It may also regulate the activity of the tachykinin-containing afferents.

Coexistence of calcitonin gene-related peptide- and substance P-like immunoreactivity in retrogradely labeled superior spermatic neurons in the dog

Coexistence of calcitonin gene-related peptide- and substance P-like immunoreactivity in retrogradely labeled superior spermatic neurons in the dog

Coexistence of calcitonin gene-related peptide- and substance P-like immunoreactivity in retrogradely labeled superior spermatic neurons in the dog

The coexistence of calcitonin gene-related peptide (CGRP) and substance P (SP) was determined in primary afferent neurons of the superior spermatic nerve of the dog. Testicular afferent neurons were visualized by retrograde labeling with the fluorescent dye fast blue. CGRP-like immunoreactivity (LI) was found in about 80% of testicular L1 and L2 dorsal root ganglion cells, and 81% of CGRP-positive neurons also contained SP. Conversely, SP-LI was found in 66% of testicular afferents, and 96% of SP-positive neurons simultaneously contained CGRP. Both CGRP- and SP-LI were observed in the whole size range of the testicular afferent neurons. No significant difference in the diameter was detected between CGRP- and SP-positive testicular afferent neurons. In contrast, the diameter of SP-positive cells was significantly smaller than that of CGRP-positive cells in the whole population of dorsal root ganglion (DRG) neurons. Compared to skin and muscle afferents, a larger population of the testicular afferents contain these peptides. This is considered to be one of the characteristic features of visceral afferents. Coexistence of CGRP and SP in testicular afferent neurons suggests a close functional relationship between these two neuropeptides in the sensory nervous system.

Calcitonin gene-related peptide and substance P in the pharynx and lung of the bullfrog, Rana catesbeiana.

Indirect double immunofluorescence labelling in the pharynx and lung of the bullfrog, Rana catesbeiana, demonstrated the occurrence, distribution, and coexistence of two neuropeptides. In the pharynx, immunoreactive calcitonin gene-related peptide (CGRP) and substance P (SP) were localized in nerve fibers distributed within and just beneath the ciliated epithelium. In the lung, CGRP and SP were localized in nerve fibers in five principal locations: 1) within the smooth muscle layer in the interfaveolar septa; 2) in the luminal thickened edges of the septa; 3) around the pulmonary vasculature; 4) within, and 5) under the ciliated epithelium. Within the smooth muscle layer in the septa, luminal thickened septa, and around blood vessels, almost all fibers showed coexistence of CGRP and SP. Within and just beneath the ciliated epithelium in the thickened septa, all fibers showed coexistence of CGRP and SP. No immunoreactivity for vasoactive intestinal polypeptide, neuropeptide Y, galanin, somatostatin, FMRFamide, and leucine- and methionine-enkephalins was detected in the nerve fibers within the larynx and the lung. Together with our previous data, the present findings suggest that peptidergic mechanisms are involved in the regulation of amphibian respiratory systems throughout their life.

Coexistence of calcitonin gene-related peptide and choline acetyltransferase in EEL efferent neurons

We applied choline acetyltransferase, (ChAT) and calcitonin gene-related peptide (CGRP) immunocytochemistry to the efferent neurons that innervate the lateral line and the ear of the eel. Strong immunoreactivity to the ChAT antiserum was observed in neurons located within the octavolateralis efferent nucleus that could be distinguished, on the basis of their form, location and dendritic organization, from the ChAT-immunopositive motoneurons of the adjacent facial motor nucleus. Both facial motoneurons and efferent neurons were found to be immunopositive for CGRP, although the reaction was always stronger in the motoneurons. Double labelling experiments established the presence of both ChAT and CGRP in many efferent neurons. The results are evidence that cholinergic efferent neurons supplying end organs of different modalities may also produce calcitonin gene-related peptide.

Immunohistochemical coexistence of calcitonin gene-related peptide and substance P in the nerve fibers of the internal gills of bullfrog ( Rana catesbeiana) larvae

A double-labelling immunofluorescence study of the internal gills of larval bullfrogs revealed coexistence of calcitonin gene-related peptide (CGRP) and substance P (SP) in the same nerve fibers in the gill tufts. More than about 95% of CGRP fibers showed coexistence of SP, although some fibers contained CGRP without SP. In the branchial muscle, all CGRP fibers demonstrated coexistence of SP. These findings suggest that both CGRP- and SP-containing fibers may use both peptides as neuromodulators of active ion transport and gill movements.

Coexistence of calcitonin gene-related peptide and galanin immunoreactivity in female rat pelvic and lumbosacral dorsal root ganglia

Coexistence of immunoreactivity for calcitonin gene-related peptide (CGRP) and galanin (GAL) was examined in varicose nerve endings in female rat pelvic paracervical ganglia (PG) and in perikarya of lumbosacral dorsal root ganglia (DRG). Varicose peptide-containing nerves were closely adjacent to somata of neurons in the PG, certain somata being virtually surrounded by immunoreactive varicosities. Some nerve endings were immunoreactive for either CGRP or GAL; in others, immunoreactivity for CGRP and GAL coexisted. Likewise, many perikarya in DRG were CGRP immunoreactive, fewer were GAL immunoreactive, and in some immunoreactivity for CGRP and GAL coexisted. The results suggest there are subpopulations of neuropeptide-containing sensory nerve endings in the PG; some contain CGRP, some contain GAL, and in some CGRP and GAL coexist. These substances contained in sensory nerve endings could have important roles in pelvic ganglionic functions.

Different types of calcitonin gene-related peptide-immunoreactive neurons in the guinea-pig stellate ganglion as revealed by triple-labelling immunofluorescence

A triple-labelling immunofluorescence technique was used to study the patterns of coexistence of calcitonin gene-related peptide (CGRP), neuropeptide Y (NPY), and tyrosine hydroxylase (TH) in the guinea-pig stellate ganglion. CGRP-immunoreactive postganglionic neurons, which all were of the non-catecholaminergic type, could be divided into NPY-positive and NPY-negative populations. Sweat glands, which are a known target of CGRP-immunoreactive sympathetic neurons, exhibited exclusively fibers of the CGRP+/NPY- phenotype. Lack of coexistence of CGRP- and NPY-immunoreactivity was also observed in nerves within skeletal muscle, brown adipose tissue and hairy skin. The findings document a previously unknown diversity of CGRP-immunoreactive postganglionic sympathetic neurons, and indicate the existence of an addition, presently unidentified target of non-catecholaminergic, CGRP-immunoreactive sympathetic neurons.

Ultrastructural and Histochemical Study of the Neuromuscular Junctions in the Denervated Intrinsic Laryngeal Muscle of the Cat

Morphological changes and acetylcholinesterase (AchE) activity in the neuromuscular junctions (NMJ) of the normal and denervated posterior cricoarytenoid muscle (PCA muscle) of the cat were studied. Two days after denervation, the nerve terminals at the NMJ had almost disappeared. Six weeks after denervation, intensity of AchE activity at the former junctional site (FJS) was unchanged histochemically. At this stage, primary synaptic clefts were distorted and the Schwann's cells covered the FJS. Fourteen weeks after denervation, AchE activity at the FJS had decreased in contrast to that of the non-affected side. Calcitonin gene-related peptide (CGRP) was also investigated at the NMJ of the normal PCA muscles immunocytochemically. The present study shows that CGRP coexists with Ach in the nerve terminals of the PCA muscles and may be involved in the regulation of the contractile function of the intrinsic laryngeal muscle.

Coexistence of calcitonin gene-related peptide and enkephalin in single neurons of the lateral superior olivary nucleus of the guinea pig that project to the cochlea as lateral olivocochlear system

Immunocytochemical double staining analysis revealed that some of the calcitonin gene-related peptide-containing neurons in the lateral olivary nucleus contained enkephalin in the guinea pig.

Coexistence of Calcitonin gene-related peptide and Enkephalin in Single Neurons of the Lateral Superior Olivary Nucleus of the guinea pig that project to the Cochlea as Lateral Olivocochlear System

Coexistence of Calcitonin gene-related peptide and Enkephalin in Single Neurons of the Lateral Superior Olivary Nucleus of the guinea pig that project to the Cochlea as Lateral Olivocochlear System

Tachykinins and calcitonin gene-related peptide: co-existence in sensory nerves of the nasal mucosa and effects on blood flow.

The presence and co-existence of calcitonin gene-related peptide (CGRP)- and substance P (SP)-like immunoreactivity (-LI) in sensory neurons of the nasal mucosa and trigeminal ganglion in several vertebrate species, including man, were established using immunohistochemistry. In the nasal mucosa the CGRP- and SP-immunoreactive (IR) nerve fibers were localized within the epithelium, around arteries, arterioles, venules, venous sinusoids and close to exocrine elements, mainly ducts. Double-staining experiments revealed that the CGRP-LI-containing nerve profiles and cell bodies also contained SP-LI. In the pig, CGRP- and SP-IR fibers were also detected in the maxillary portion of the trigeminal nerve and around the sphenopalatine artery and vein, as well as around the nasal dorsal vein. The nasal mucosal content of CGRP-LI, as determined by radioimmunoassay, was almost 5-fold higher in the pig and guinea pig compared to man. The nasal CGRP-IR nerves disappeared after capsaicin pretreatment in the guinea pig. In the cat, local intra-arterial infusions of capsaicin, SP, neurokinin A (NKA), neuropeptide K (NPK) and CGRP caused a concentration-dependent increase in nasal blood flow. CGRP caused a longer-lasting vasodilatation than the tachykinins. In conclusion, the morphological findings of co-localization of CGRP-LI and SP-LI in capsaicin-sensitive nerve fibers of the nasal mucosa and trigeminal ganglia of different species including man, coupled with the in vivo description of the high vasodilator potency of CGRP and tachykinins, imply co-release of several vasoactive agents upon activation of the nasal sensory nerves. Furthermore, the similarity of the morphological findings among the different species indicates that experimental data from animals may reflect the existence of similar mechanisms in humans.

Inhibition of gastric and pancreatic secretion in dogs by CGRP: role of somatostatin

The coexistence of calcitonin gene-related peptide (CGRP) and somatostatin (SS) within the stomach and pancreas and the potent inhibitory effects of both peptides on exocrine secretions from these organs suggest that they are functionally related. To assess the potential role of SS in the mediation of CGRP action, the effects of intravenous human CGRP (64, 132, and 264 pmol.kg-1.h-1) and somatostatin-14 (SS-14; 100, 400, and 800 pmol.kg-1.h-1) on plasma levels of SS immunoreactivity (SS-IR) and on pentagastrin-stimulated gastric and pancreatic secretion were compared in conscious dogs. CGRP caused significant inhibition of gastric acid (85-102%), pancreatic protein (63-86%), and pancreatic bicarbonate (74-89%) outputs and a simultaneous dose-related rise (40-102 fmol/ml) in plasma SS-IR. Cessation of CGRP infusion resulted in prompt return of plasma SS-IR to basal levels and an increase in gastric and pancreatic secretion. Although CGRP is a potent releasor of SS into the circulation, its inhibitory action on gastric acid secretion cannot be explained solely by a rise in plasma SS-IR. In the pancreas, in contrast to the stomach, inhibition appears to be more closely related to a rise in circulating level of SS-IR.

The coexistence of neuropeptides in feline sensory neurons

The coexistence of immunoreactivity to the peptides substance P, bombesin, calcitonin gene-related peptide and somatostatin has been determined in single, lumbar and sacral dorsal root ganglion cells in the cat. Colchicine pretreated L7 and S1 dorsal root ganglia were embedded in wax and cut into 5 μm sections. Groups of four, serially adjacent sections were reacted with antisera to one of four peptides using avidin-biotin immunocytochemistry. It was thus possible to determine the coincidence of the four peptides in single cell bodies by examining the immunoreactivity in a ganglion cell in one section and then locating the same cell in three adjacent sections. As a comparison, this procedure was repeated on a different population of ganglion cells using antiserum to substance P, bombesin and calcitonin gene-related peptide only. The results indicate that different combinations of three or four peptides may occur in single, small diameter sensory neurons in the cat. It would appear that immunoreactivity to bombesin and/or calcitonin gene-related peptide coexists with immunoreactivity to substance P in some dorsal root ganglion cells. However, immunoreactivity to each of these peptides was also found to occur alone in single cells. Immunoreactivity to calcitonin gene-related peptide but not to the other three peptides was found to occur in some medium-sized cell bodies (up to 70 μm). Somatostatin-like immunoreactivity was found to have a high level of coexistence with substance P-like immunoreactivity in cells which contained immunoreactivity to these two peptides only. Immunoreactivity to all the four peptides tested was found to occur in 18–26% of ganglion cells which contained at least one peptide.

Existence and coexistence of calcitonin gene-related peptide, vasoactive intestinal polypeptide- and somatostatin-like immunoreactivities in spinal cord motoneurons of developing embryos and post-hatch chicks

By use of immunocytochemical methods, it is shown that immunoreactive calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and somatostatin (SOM) are present in motoneurons in the chicken spinal cord. While CGRP-like immunoreactivity (LI) is present in numerous motoneurons both before and after hatching, SOM- and VIP-LI markedly decline at the end of the embryonic period. Evidence is also provided for coexistence of some of these peptides in certain spinal motoneurons.

Immunoreactivity for calcitonin gene-related peptide in neuroepithelial bodies of the newborn cat.

Immunoreactivity for calcitonin gene-related peptide is demonstrated for the first time in neuroepithelial bodies in the lung of newborn cats after Bouin fixation and embedding in paraffin. The intense staining clearly identifies these bodies at the level of bronchioli and alveoli. Occasionally, single neuroepithelial endocrine cells, displaying immunoreactivity for calcitonin gene-related peptide are observed. In the kitten lung, identification and localization of neuroepithelial bodies after immunocytochemical staining for calcitonin gene-related peptide are superior to the analysis based on other techniques, i.e., the argyrophilic reaction, periodic acid Schiff-lead hematoxylin method, and immunocytochemical staining for serotonin. The serial-section technique revealed that in neuroepithelial bodies of the newborn kitten lung, immunoreactivity for calcitonin gene-related peptide coexists with immunoreactivity for serotonin in individual cells. The functional significance of the calcitonin gene-related peptide in neuroepithelial bodies remains to be elucidated.

Immunohistochemical evidence for the coexistence of calcitonin gene-related peptide and glutamate decar☐ylase-like immunoreactivities in the Purkinje cells of the rat cerebellum

Immunohistochemical staining of alternate consecutive sections revealed numerous glutamate decar☐ylase-like immunoreactive Purkinje cells that also contained calcitonin gene-related peptide in the rat cerebellum.