Localization Of Calcitonin Gene-related Peptide Research Articles

Biodegradable Bisvinyl Sulfonemethyl-crosslinked Gelatin Conduit Promotes Regeneration after Peripheral Nerve Injury in Adult Rats

In our previous study, we found that gelatin-based materials exhibit good conductivity and are non-cytotoxic. In this study, gelatin was cross-linked with bisvinyl sulfonemethyl (BVSM) to fabricate a biodegradable conduit for peripheral nerve repair. First, BVSM on the prepared conduit was characterized to determine its mechanical properties and contact angle. The maximum tensile strength and water contact angle of the gelatin-BVSM conduits were 23 ± 4.8 MPa and 74.7 ± 9°, which provided sufficient mechanical strength to resist muscular contraction; additionally, the surface was hydrophilic. Cytotoxicity and apoptosis assays using Schwann cells demonstrated that the gelatin-BVSM conduits are non-cytotoxic. Next, we examined the neuronal electrophysiology, animal behavior, neuronal connectivity, macrophage infiltration, calcitonin gene-related peptide localization and expression, as well as the expression levels of nerve regeneration-related proteins. The number of fluorogold-labelled cells and histological analysis of the gelatin-BVSM nerve conduits was similar to that observed with the clinical use of silicone rubber conduits after 8 weeks of repair. Therefore, our results demonstrate that gelatin-BVSM conduits are promising substrates for application as bioengineered grafts for nerve tissue regeneration.

The localization of calcitonin gene-related peptide in the human trigeminal ganglion and masseter muscle.

The localization of calcitonin gene-related peptide (CGRP) is similar to that of a neurotransmitter which indicates masticatory muscle pain in the area of the masseter fascia. CGRP is released from the trigeminal ganglion (TG). The aim of this study was to analyze the distribution of CGRP in the fascia of the masseter muscle (FMM) and TG in a morphometric manner, with respect to the location and density of CGRP-immunopositive reaction fiber (CGRP-IRF). A higher number of the CGRP-IRF were mainly found located around elongated blood vessels and small nerves on the origin side of the middle zone FMM in the O group (presented with occlusion). In the sectional histochemical analysis of the O group, the CGRP-IRF were clearly detected in oval vessels, large elongated vessels and large nerves in contrast with that of the Non-O group (presented with no occlusion) samples. The number of CGRP-immunopositive ganglion cells (CGRP-IPGCs) in the O group mandibular nerve division was higher than that of other divisions. A reduction of the CGRP-IRF numbers were found in the no-loading groups. The characterization of these locations of CGRP-IPGCs can also provide useful data for the understanding of myofascial pain syndrome of the masseter muscle (MM).

Developmental localization of calcitonin gene-related peptide in dorsal sensory axons and ventral motor neurons of mouse cervical spinal cord

Calcitonin gene-related peptide (CGRP) is a 37-amino-acid neuropeptide, synthesized by alternative splicing of calcitonin gene mRNA. CGRP is characteristically distributed in the nervous system, and its function varies depending on where it is expressed. To reveal developmental formation of the CGRP network and its function in neuronal maturation, we examined the immunohistochemical localization of CGRP in the developing mouse cervical spinal cord and dorsal root ganglion. CGRP immunolabeling (IL) was first detected in motor neurons on E13, and in ascending axons of the posterior funiculus and DRG neurons on E14. CGRP-positive sensory axon fibers entered Laminae I and II on E16, and Laminae I through IV on E18. The intensity of the CGRP-IL gradually increased in both ventral and dorsal horns during embryonic development, but markedly decreased in the ventral horn after birth. These results suggest that CGRP is expressed several days after neuronal settling and entry of sensory fibers, and that the CGRP network is formed in chronological and sequential order. Furthermore, because CGRP is markedly expressed in motor neurons when axons are vastly extending and innervating targets, CGRP may also be involved in axonal elongation and synapse formation during normal development.

Localization of Calcitonin Gene-Related Peptide in the Inner Ear of the Rat

Localization of Calcitonin Gene-Related Peptide in the Inner Ear of the Rat

Early potential impairment of renal sensory nerves in streptozotocin-induced diabetic rats: role of neurokinin receptors

Electrophysiological studies in the mammalian kidney have identified two major classes of sensory receptors of the afferent renal nerves; chemoreceptors (CR) and mechanoreceptors (MR). The localization of calcitonin gene-related peptide (CGRP) and substance P (SP) in these renal pelvic sensory neurons provides an anatomical basis for a possible functional interaction between the two neuropeptides and SP receptor. The present study was performed to examine the possible changes in the responsiveness of renal sensory SP and CGRP receptors in rats with streptozotocin (STZ)-induced diabetes mellitus. Due to the crucial role of renal pelvic SP and CGRP receptors in the activation of renal sensory neurons by various stimuli, we examined whether the responsiveness of MR or CR activation and the dorsal root ganglia content of neuropeptides and neurokinin 1 receptors (NK(1)R) were altered in diabetic rats compared with non-diabetic rats. Afferent renal nerve activity (ARNA) was recorded from the peripheral portion of the cut end of one renal nerve branch placed on a bipolar silver wire electrode. T(13) dorsal root ganglia (DRG) immunoreactivity was performed to NK(1)R, SP and CGRP. The results of the current study confirmed that the stimulation of renal MR and CR elicited a renorenal reflex response, and that the renal pelvic administration of SP and CGRP increased ipsilateral ARNA and contralateral urinary sodium excretion with no changes in arterial pressure. We also found a decrease in NK(1)R expression followed by an increase in SP and CGRP levels in the DRG of diabetic rats. The ARNA response, produced by renal pelvic MR and CR stimulation, was found to be significantly attenuated in the STZ-induced diabetic model. Conclusions. These data may indicate a compensatory synthesis and/or abnormal axonal delivery of neurokinins from the cell body to synaptic portions of the neuron as the underlying reason for attenuated ARNA in renal sensory neurons of diabetic rats.

Unique Matrix Structure in the Rough Endoplasmic Reticulum Cisternae of Pseudoachondroplasia Chondrocytes

Mutations in cartilage oligomeric matrix protein (COMP) cause two skeletal dysplasias, pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (MED/EDM1). Because COMP exists as a homopentamer, only one mutant COMP subunit may result in an abnormal complex that is accumulated in expanded rough endoplasmic reticulum (rER) cisternae, a hallmark of PSACH. Type IX collagen and matrilin-3 (MATN3), also accumulate in the rER cisternae of PSACH chondrocytes, but it is unknown how mutant COMP interacts with these proteins. The studies herein focus on defining the organization of these intracellularly retained proteins using fluorescence deconvolution microscopy. A unique matrix organization was identified in which type II procollagen formed a central core surrounded by a protein network of mutant COMP, type IX collagen, and MATN3. This pattern of matrix organization was found in multiple cisternae from single chondrocytes and in chondrocytes with different COMP mutations, indicating a common pattern of interaction. This suggests that stalling of mutant COMP and an interaction between mutant COMP and type II procollagen are initiating events in the assembly of matrix in the rER, possibly explaining why the material is not readily cleared from the rER. Altogether, these data suggest that mutant COMP initiates and perhaps catalyzes premature intracellular matrix assembly.

Immunohistochemical localization of calcitonin gene-related peptide and substance P in the rat knee cartilage at birth.

Substance P (SP) and calcitonin gene-related peptide (CGRP) have been found in the perichondrium and within the cartilage canals. It is still unknown whether they exert a direct effect on chondrocytes during joint development. We processed 28 knees of newborn Wistar rats in 7 different fashions to perform histology and immunohistochemistry studies. Positive immunoreactivity against CGRP and SP was found in the inner aspect of the perichondrium in a close contact with chondrocytes. The presence of CGRP and SP indicates the presence of nerves fibers, and precedes the development of cartilage canals. Nerve fibers may play a role in the development of synovial joints before and during the presence of cartilage canals. The presence of CGRP and SP in the cartilage at birth may be involved in the early postnatal maturation of synovial joints. It remains to be determined whether autonomic innervation is later involved in age-related degenerative joint disease.

Localization of calcitonin gene-related peptide in cardiomyocytes: comparison of neonatal and dedifferentiating cells to adult myocytes

The purpose of this study was to localize sites of calcitonin gene-related peptide binding in neonatal, freshly isolated and dedifferentiated adult cardiac myocytes in order to help us elucidate the mechanisms of action of this neuropeptides. Previous work has shown that treatment with calcitonin gene-related peptide results in dramatic changes in calcium transients, so we carried out multi-channel acquisitions of fluorescently labeled images to reveal where calcitonin gene-related protein and the l-type calcium channel were localized. Calcitonin gene-related protein was sparse and randomly distributed in rod-like adult cardiomyocytes, found in abundance in areas of the cell where striations were apparent and not where adhesion proteins predominated in dedifferentiating adult myocytes, and in a large perinuclear concentration, with some spreading into the cytoplasm in neonatal cells. Subsequent modeling demonstrated that calcitonin gene-related peptide and the l-type calcium channel protein were closely associated in each of the three myocyte types, suggesting that while the peptide has dramatic and different effects on intracellular calcium levels of the various cardiomyocytes, the action is probably via diverse mechanisms as a result of effects on different channels or pump proteins due to alterations in intracellular calcium concentrations.

Role of Capsaicin-Sensitive Nerve Fibers in Uterine Contractility in the Rat1

The possible participation of capsaicin-sensitive sensory nerves in the modulation of neurogenic contractions was studied in nonpregnant and term pregnant rat uteri. Neurogenic contractions were elicited by electric field stimulation (40 V, 1-70 Hz, 0.6 msec) in intact uteri and uteri that were previously exposed to capsaicin in vitro. In capsaicin pretreated preparations obtained both from nonpregnant and term pregnant rats, a dose-dependent increase in the amplitude of uterine contractions was detected. Prior systemic treatment of the rats with capsaicin (130 mg/kg, s.c.) abolished the effect of in vitro capsaicin administration on the amplitude of neurogenic contractions. Use of a specific antagonist of calcitonin gene-related peptide revealed that depletion of this peptide, which normally elicits uterine smooth muscle relaxation, may be responsible for the increased responsiveness of the uterus to low-frequency stimulation. Experiments on the localization of calcitonin gene-related peptide in uterine tissue specimens exposed to capsaicin revealed dose-dependent depletion of calcitonin-gene related peptide-immunoreactive nerves innervating blood vessels and the myometrium. The findings indicate that capsaicin-sensitive afferent nerves, by the release of sensory neuropeptides, significantly contribute to the modulation of uterine contractility both in nonpregnant and term pregnant rats. It is suggested that uterine sensory nerve activation may be part of a trigger mechanism leading to preterm contractions evoked by, for example, inflammation.

Subcellular distributions of calcitonin gene-related peptide (CGRP)-like immunoreactivity in the subcommissural organ of the golden hamster ( Mesocricetus auratus)

Immuno-electron microscopy specifically enhanced with silver staining has been used to demonstrate the localization of calcitonin gene-related peptide (CGRP) in the ependymocytes of the hamster subcommissural organ (SCO). Hamster SCO consists of the ependymal and hypendymal cell layers, the latter being arranged as rosette-like structure across the posterior commissure (PC) and often arranged with longitudinal axis parallel to the ventricle. All cytoplasmic regions of the ependymal and hypendymal cells were strongly stained with CGRP. In the hypendymal layer, the CGRP positive hypendymal cells were frequently in contact with local blood vessels and arranged in-groups traversing the thick portion of the PC. Ultrastructurally, the CGRP-immunoreaction products were distributed at the dilated cisternae of the rough endoplasmic reticulum (rER) and secretory granules of the ependymal and hypendymal cells. The dilated cisterna of the rER was usually concentrated in the basal region of the ependymal cells and irregular in shape. These dilated cisternae were filled with a flocculent material or finely granular substance, but hardly studded with ribosomes. Labelled secretory granules were abundant in the apical pole of the ependymal cells and discharged their contents into the third ventricle in the form of a thin layer of secretion. This CGRP positive material appeared to constitute the pre-Reissner's fiber (RF). On the basis of the present ultrastructural evidences, we proposed that ependymocytic CGRP in SCO may be synthesized and stored in the cisternae of rER, then released and incorporated into the RF in the third ventricle through the secretory granules. The abundant amount of CGRP in ependymocytes of SCO and RF in the third ventricle suggests a significant endocrine function of CGRP in hamster SCO.

Immunohistochemical Localization of Calcitonin Gene-Related Peptide in the Human Gastric Mucosa

Background/Aim: There have been only a few studies on the distribution of calcitonin gene-related peptide (CGRP) in the human stomach, in which it was stated that CGRP fibers are rare in that organ. The aim of the present study was to investigate the immunohistochemical localization of CGRP in the human gastric mucosa obtained by endoscopic biopsy from patients with gastric ulcers. Methods: Immunohistochemistry was carried out according to the indirect immunoperoxidase method using an anti-human CGRP antibody. Biopsies were taken from the ulcer margin in 18 patients (age 37–78, average 57.4 years) and from two endoscopically normal portions (antrum and body) in 7 other patients (age 36–65, average 51.0 years). One biopsy specimen was obtained from each portion. Results: Twelve of the eighteen biopsy specimens from the ulcer margin, 6 of the 7 biopsy specimens from normal portions of the antrum and 3 of the 7 biopsy specimens from normal portions of the body showed CGRP-immunoreactive staining. Intense staining was more marked in the specimens from the ulcer margin compared to those of the normal portions. Conclusions: CGRP immunoreactivity was observed in the human gastric mucosa in considerable abundance, and it is presumed that CGRP might participate in a restoration mechanism of the ulcer.

Localization of calcitonin gene-related peptide within the genitofemoral nerve in immature rats

Background/Purpose : Calcitonin gene-related peptide (CGRP) has been proposed to influence migration and testicular descent by release from the genitofemoral nerve. The site of CGRP within the nerve has been controversial, with conflicting views on whether CGRP is synthesised and released from the motor nerves. Methods : The genitofemoral nerve (GFN) was retrogradely labelled by fluorescent dye (DAPI) in 25 Sprague-Dawley rats (days 5, 16, and 31, n = 8 in each group; day 35, n = 1). Spinal cords and dorsal root ganglia (DRG) were removed two to three days later and sectioned for immunofluorescence. Substance P and CGRP-containing cells were labelled with fluorescein-linked antibodies. Specimens were examined by double fluorescence to identify cells with both markers. Results : The motor nucleus of the GFN contained 119 cells on day 7 and 284 cells by days 19 through 34. A prominent band of CGRP-containing fibers, arising from the dorsal horn, synapsed with the GFN motor nucleus itself. CGRP-labelled GFN cells were found in the DRG by double labelling. Conclusions : CGRP from the GFN may affect gubernacular migration by release from the sensory nerves, rather than motor nerves as previously thought. The GFN motor nucleus receives CGRP-containing innervation from the dorsal horn, which may form part of the cremasteric reflex.

Light microscopic localization of calcitonin gene-related peptide in the normal feline trigeminal system and following retrogasserian rhizotomy.

Calcitonin gene-related peptide (CGRP) is a neuropeptide that has been implicated in the transmission and modulation of primary afferent nociceptive stimuli. In this study, we describe the light microscopic distribution of CGRP immunoreactivity (IR) within the feline trigeminal ganglion and trigeminal nucleus of normal adult subjects and in subjects 10 and 30 days following complete retrogasserian rhizotomy. Within the trigeminal ganglion of normal subjects, cell bodies and fibers showed CGRP-IR, whereas immunoreactive fibers were rare in the central root region. Within the normal spinal trigeminal and main sensory nuclei, CGRP-IR was seen to form a reproducible pattern that varied between the different nuclei. Following rhizotomy, most, but not all, of the CGRP-IR was lost from the spinal trigeminal and main sensory nuclei, except in regions where the upper cervical roots and cranial nerves VII, IX and X project into the trigeminal nucleus. The pattern seen at 10 days contained more CGRP-IR than that seen at 30 days and suggests that degenerating fibers still show CGRP-IR. In contrast to the decrease seen in the nuclei after rhizotomy, examination of the central root that was still attached to the trigeminal ganglion showed an increase in CGRP-IR within fibers, some of which ended in growth conelike enlargements. Rhizotomy induced a dramatic increase in CGRP-IR within trigeminal motoneurons and their fibers, which was strongest 10 days after rhizotomy and weaker at 30 days, which was still stronger than normal. These results indicate that the majority of CGRP-IR found in the trigeminal nucleus originates from trigeminal primary afferents and that an upregulation of CGRP-IR occurs in trigeminal motoneurons and in regenerating fibers in the part of the central root that was still attached to the ganglion. In addition, the persistence of CGRP-IR fibers in the trigeminal nucleus provides one possible explanation for the preservation of pain in humans following trigeminal rhizotomy.

Localization of calcitonin gene-related peptide binding sites in the eye of different species.

The localization of calcitonin gene-related peptide (CGRP) binding sites in the eye of monkey, pig, cat and guinea pig was studied by autoradiography. Specific binding of CGRP was found in ciliary muscle, ciliary processes and limbal conjunctiva in all tested species. Furthermore, specific binding sites of CGRP was found in the choroidea of monkey, pig and guinea pig, in the iris of pig, cat and guinea pig, in the retina of pig and in the anterior chamber angle of cat. The number of specific binding sites varied depending on the tissue and species. The present study shows that there are specific binding sites of CGRP in the eye of monkey, pig, cat and guinea pig. CGRP binding sites found in vascular system of ciliary body, choroidea and iris further demonstrates the role of CGRP as a vasoregulatory peptide. Binding sites in the ciliary muscle, in the limbal conjunctiva and in the chamber angle area may indicate a role in the regulation of ciliary muscle tone, epithelial cell regeneration and aqueous humour outflow.

Localization, identification, and action of calcitonin gene-related peptide in the frog adrenal gland.

The localization of calcitonin gene-related peptide (CGRP)-like immunoreactivity in the adrenal gland of the frog, Rana ridibunda, was examined by the indirect immunofluorescence technique. Using an antiserum directed against rat alpha-CGRP, the presence of a network of positive fibers was observed in the adrenal parenchyma. The immunoreactive material has been characterized by HPLC analysis combined with RIA quantification. The elution profile revealed the existence of a single form of CGRP exhibiting the same retention time as synthetic frog CGRP. The possible involvement of CGRP in the regulation of corticosteroid secretion was studied in vitro using a perifusion system for frog adrenal slices. Graded doses of frog CGRP (from 3 x 10(-9) to 3 x 10(-6) M) increased corticosterone and aldosterone secretion in a dose-dependent manner (ED50, 4.1 x 10(-8) M). Several other forms of CGRP, i.e. rat alpha-CGRP and beta-CGRP, and human alpha-CGRP and beta-CGRP, were also capable of enhancing steroid output, but frog CGRP was the most effective stimulator of steroidogenesis. Repeated administration of rat alpha CGRP induced a reproducible stimulation of corticosteroid secretion without any tachyphylaxis. Prolonged infusion of the peptide (3 h) caused a rapid increase in corticosteroid release, followed by a gradual decline of steroid secretion, suggesting the occurrence of a desensitization phenomenon. Rat alpha-CGRP also gave rise to a significant increase in corticosteroid release from acutely dispersed adrenal cells. These results show the presence of CGRP in fibers innervating the frog adrenal gland. The data also demonstrate that synthetic CGRP exerts a direct stimulatory effect on corticosteroid secretion. Taken together, these findings suggest that CGRP, released by nerve fibers in the adrenal tissue, can locally regulate corticosteroid secretion.

Localization of calcitonin gene-related peptide and islet amyloid polypeptide in the rat and mouse pancreas

It was previously demonstrated that the two chemically related peptides calcitonin gene-related peptide (CGRP) and islet amyloid polypeptide (IAPP) both occur in the pancreas. We have now examined the cellular localization of CGRP and IAPP in the rat and the mouse pancreas. We found, in both the rat and the mouse pancreas, CGRP-immunoreactive nerve fibers throughout the parenchyma, including the islets, with particular association with blood vessels. CGRP-immunoreactive nerve fibers were regularly seen within the islets. In contrast, no IAPP-immunoreactive nerve fibers were demonstrated in this location. Furthermore, in rat islets, CGRP immunoreactivity was demonstrated in peripherally located cells, constituting a major subpopulation of the somatostatin cells. Such cells were lacking in the mouse islets. IAPP-like immunoreactivity was demonstrated in rat and mouse islet insulin cells, and, in the rat, also in a few non-insulin cells in the islet periphery. These cells seemed to be identical with somatostatin/CGRP-immunoreactive elements. In summary, the study shows (1) that CGRP, but not IAPP, is a pancreatic neuropeptide both in the mouse and the rat; (2) that a subpopulation of rat somatostatin cells contain CGRP; (3) that mouse islet endocrine cells do not contain CGRP; (4) that insulin cells in both the rat and the mouse contain IAPP; and (5) that in the rat, a non-insulin cell population apparently composed of somatostatin cells stores immunoreactive IAPP. We conclude that CGRP is a pancreatic neuropeptide and IAPP is an islet endocrine peptide in both the rat and the mouse, whereas CGRP is an islet endocrine peptide in the rat.

Calcitonin gene-related peptide in nucleus ambiguus motoneurons in rat: viscerotopic organization.

Calcitonin gene-related peptide has been reported in the rat nucleus ambiguus. This nucleus comprises a dorsal division that is the source of special visceral efferents innervating the striated muscle of the upper alimentary tract and a ventral division supplying general visceral efferents primarily to the heart. The distribution of calcitonin gene-related peptide immunoreactive neurons in the two divisions was determined by using a combination of immunocytochemical techniques and fluorescent retrograde tracing. In 22 rats, injections of Fluoro-Gold were made into either the supranodosal vagus nerve, palatopharynx, larynx, esophagus, or heart. Following colchicine injection, medullary sections were processed immunocytochemically for calcitonin gene-related peptide. Injection of Fluoro-Gold into the supranodosal vagus resulted in prominent labeling of neurons in the dorsal and ventral divisions of the nucleus ambiguus. The majority of fluorescent labeled neurons in the dorsal division were found to be immunoreactive for calcitonin gene-related peptide, while those labeled neurons in the ventral division were unreactive for the peptide. With esophageal, and palatopharyngeal and cricothyroid injections, many fluorescent labeled neurons that were immunoreactive for calcitonin gene-related peptide were found respectively in the compact and semicompact formations of the dorsal division. In contrast, injections of the heart resulted in fluorescent labeled neurons, which were unreactive for calcitonin gene-related peptide, localized to the external formation. The results demonstrate that calcitonin gene-related peptide immunoreactive neurons are localized entirely to the dorsal division of the nucleus ambiguus and that all striated muscular areas of the alimentary tract are innervated by calcitonin gene-related peptide containing motoneurons. The localization of calcitonin gene-related peptide to vagal motoneurons also known to contain acetylcholine and the increase in acetylcholine receptor synthesis caused by this peptide suggest that calcitonin gene-related peptide acts as a cotransmitter with acetylcholine in special visceral efferent vagal motoneurons.

Immunohistochemical localization of calcitonin gene-related peptide and cotransmitters in a subpopulation of post-ganglionic neurons in the porcine inferior mesenteric ganglion.

Applying double-fluorescence immunohistochemistry, adrenergic and non-adrenergic postganglionic sympathetic neurons, in the porcine inferior mesenteric ganglion (IMG) are subdivided according to size and cotransmitter content. Calcitonin gene related peptide (CGRP)-immunoreactive (IR) neurons are demonstrated to belong to the non-adrenergic, i.e. tyrosine hydroxylase- and DOPAmine-beta-hydroxylase-(D beta H)-negative subpopulation of postganglionic perikarya. Virtually all of the CGRP-IR postganglionic neurons exhibit colocalization with somatostatin (SOM), and, some of them with neuropeptide tyrosine (NPY). Additionally, NPY-, SOM-, and NPY/SOM-IR subpopulations of adrenergic and non-adrenergic neurons are observed. CGRP-immunoreactivity is seen in dense networks of intraganglionic varicose nerve fibres, adjacent to the TH- and SOM-IR neurons. NPY-IR perikarya are sparsely supplied by CGRP-IR fibres. SOM- and NPY-IR nerve fibres also exist in the inferior mesenteric ganglion. The functional relevance of CGRP-IR postganglionic neurons, as well as target organs of these neurons remain to be elucidated.

Anatomic distribution of autonomic neural tissue in the developing dog heart: II. Nonadrenergic noncholinergic innervation by calcitonin gene‐related peptide‐immunoreactive tissue

We used immunocytochemical localization of calcitonin gene-related peptide (CGRP) to trace the ontogenesis and anatomic distribution of this component of nonadrenergic noncholinergic (NANC) innervation in fetal, neonatal, and mature canine hearts and autonomic ganglia which control cardiac function. Rare varicose CGRP-immunoreactive nerve processes were present in the heart during late gestation. Abundant CGRP-immunoreactive neural tissue in the neonate suggested a burst of NANC innervation around birth. Neonatal, 1-, and 2-month-old animals all had many varicose individual nerve processes in addition to processes within bundles; however, the density of all CGRP-immunoreactive tissue appeared to decrease during this stage of development. Similarly, there were relatively more varicose stained nerve processes in the epicardial ganglia and numerous CGRP-immunoreactive cells and smooth nerve processes in the stellate ganglia of the neonate, as compared with older animals. In the mature animal CGRP-immunoreactive neural tissue in the heart was more sparse and largely confined to heterogeneous nerve bundles in the epicardium. The extramural coronary arteries were virtually the only site of innervation by individual nerve processes; CGRP-immunoreactive neural tissue was not found adjacent to working cardiac muscle fibers. At all developmental stages, the area of the sinoatrial node was the primary focus of CGRP innervation, although the atrioventricular nodal region was also preferentially innervated. In general, the atria contained more CGRP-immunoreactive tissue than the ventricles, which were only sparsely innervated. The perinatal peak in density of CGRP-immunoreactive neural tissue with subsequent decline to reach the adult pattern suggests a developmental role for NANC innervation in the dog heart.

Immunohistochemical localization of calcitonin gene-related peptide in the rat eye: Response to capsaicin pretreatment

Immunohistochemical localization of calcitonin gene-related peptide in the rat eye: Response to capsaicin pretreatment