Development of substance P-immunoreactive neurons in cranial sensory ganglia of the rat

Substance P-like immunoreactivity has been observed in fetal and adult cranial sensory ganglia. It first appears at day 16 of gestation in sensory neurons of trigeminal, superior-jugular, petrous and nodose ganglia, as well as in the autonomic myenteric plexus, and at day 17 in cervical dorsal root ganglion cells. Substance P immunoreactivity can be visualized much earlier (day 12) in the central nervous system. The ganglionic immunoreactivity subsequently increases during fetal life but drops at birth. The reactive material is first diffuse, then slowly becomes granular, and is mostly concentrated in coarse perinuclear inclusions in adult sensory neurons. Most substance P-positive neurons in trigeminal and superior-jugular ganglia are small, but medium-sized and large positive neurons are also observed in the trigeminal, petrous and nodose ganglia.Our observations give a precise picture of the development of substance P immunoreactivity in sensory neurons and are in general agreement with previous reports on some fetal and adult rat sensory ganglia. They indicate that in the rat, maturation of peripheral substance P-containing sensory neurons is slower than that of central substance P neurons or equivalent sensory neurons in other species. The examination of fetal material allows the observation of numerous immunoreactive sensory neurons which cannot be visualized after birth. We hypothesize a possible different embryonic origin (neural crest or placodal) for small nociceptive and larger substance P-containing neurons in rat cranial sensory ganglia.     

Substance P-like immunoreactivity in developing cranial parasympathetic neurons of the rat

Substance P-like immunoreactivity (SPLI) has been observed in cell bodies of fetal cranial parasympathetic ganglia of rat. It first appears at day 16 of gestation at the same time as in cranial sensory ganglia. From day 17 to 21, SPLI neurons constitute most, if not all, submandibular-sublingual and intralingual ganglia, they form 30–40% of otic and pterygopalatine ganglia and numerous such neurons are found in the myenteric plexus of the esophagus as well as in pharyngeal and buccal walls. The immunoreactive material is thinly granular, and its appearance does not change with prenatal development. The immunoreactivity in cell bodies of parasympathetic ganglia decreases at the end of the gestational period, and cannot be evidenced any more in most cells of normal adult ganglia. However, the corresponding SPLI fibers remain intensely immunoreactive. When grafted to rat irides, which have been chemically depleted of intrinsic SPLI fibers, submandibular, otic and pterygopalatine ganglia from pre- or postnatal rats rapidly produce a large amount of SPLI fibers on the iris mimicking the pattern of sensory innervation. This proves the presence of SPLI neurons in adult parasympathetic ganglia, at least in experimental conditions.This study of fetuses and grafts demonstrates the existence of neurons in SPLI parasympathetic cranial ganglia which has been underestimated or ignored previously as a result of observations on adult ganglia. The very large proportion of SPLI neurons in the ganglia of the salivary gland might be of importance for the interpretation of experimental studies on the control of salivation. The presence of SPLI in all three types of peripheral ganglia, sensory, sympathetic and parasympathetic, raises the question of its functional significance in the different compartments of the peripheral nervous system.     

Retrograde axonal transport of horseradish peroxidase in peripheral autonomic nerves.

An exogeneous marker protein, horseradish peroxidase (HRP) was used to race peripheral autonomic pathways in adult guinea pigs and cats. Small doses of HRP were injected into various organs and after a brief survival period, HRP activity appeared in the perikarya of autonomic neurons that supplied each injection site. After injection of HRP into the anterior chamber of the eye, reaction product was detected in the postganglionic sympathetic neurons of the superior cervical sympathetic ganglion. In another experiment, HRP reaction product was found in the cell bodies of the preganglionic sympathetic neurons that supply the adrenal medulla. These were located in the lateral gray column of the spinal cord at T6 and T7 segmental levels. Reaction product appeared in intramural postganglionic parasympathetic neurons close to an injection site in the wall of the urinary bladder and in a similiar situation in Meissner's ganglia of the ileum. Following injection into the walls of the stomach and ileum, HRP labelled cells were detected in the nodose ganglion of the vagus and in preganglionic parasympathetic neurons in the dorsal motor nucleus of this nerve. After injection into the subepicardial tissue of the heart, reaction product appeared in the stellate ganglion and also in an upper thoracic dorsal root ganglion. These data suggest that HRP is taken up by peripheral autonomic nerves of all types, and then undergoes rapid retrograde axonal transport to the perikaryon. It appears, therefore, that HRP may be useful in tracing both motor and sensory peripheral autonomic pathways.     

VRL-1 immunoreactivity in the rat cranial autonomic ganglia.

Immunohistochemistry for VRL-1, a newly cloned capsaicin-receptor homologue, was performed on the rat cranial autonomic ganglia. The immunoreactivity (ir) was detected in the majority of neurones in the pterygopalatine (66%) and submandibular ganglia (68%). In the tongue and carotid body, parasympathetic neurones contained VRL-I ir. In the superior cervical ganglion, only 2% of postganglionic sympathetic neurones showed the immunoreactivity. VRL-1-ir nerve endings could not be detected in their peripheral tissues. These findings may suggest that VRL-1 has functions within neuronal cell bodies of the cranial autonomic ganglia.     

Parkinson's Disease: Distribution of Lewy Bodies in the Peripheral Autonomic Nervous System

This report concerns the presence and distribution of Lewy bodies in the peripheral nervous system of three patients with Parkinson's disease and ten age-matched nonparkinsonian individuals. We examined the paravertebral and celiac ganglia as representatives of the sympathetic system; the submandibular ganglion as representative of the parasympathetic system; the esophagus, stomach, duodenum, jejunum, ileum and colon as representatives of the enteric system, and the trigeminal and dorsal spinal ganglia as representatives of the somatic sensory system. Lewy bodies were observed in the peripheral sympathetic and enteric systems of two of the three patients with Parkinson's disease. These two patients had numerous Lewy bodies in the submandibular ganglia. The third patient did not have Lewy bodies in the peripheral autonomic nervous system, nor did the ten control individuals studied. Lewy bodies were not detected in the somatic sensory system of any of the three parkinsonian patients. Our findings indicate that the peripheral autonomic system, including the parasympathetic system is affected in Parkinson's disease.     

Somatostatin-like immunoreactivity in primary afferents of the medial articular nerve and colocalization with substance P in the cat

The proportion of somatostatin-containing dorsal root ganglion cells innervating the knee joint of the cat via the medial articular nerve was determined by using retrograde labeling with fast blue and immunohistochemistry. Immunoreactivity was found in 8.6% of labeled cell bodies. In colchicine-treated ganglia, the proportion increased to 16.8%. Only small and intermediate-sized perikarya showed somatostatin-like immunoreactivity, indicating that this neuropeptide is synthesized predominantly in primary afferent units with unmyelinated sensory axons but may also be present in primary afferents with thinly myelinated sensory fibers. Colchicine treatment had no influence on the cell size distribution. Colocalization of somatostatin with substance P was determined by comparing the proportions of immunopositive dorsal root ganglion cells after incubation with antibodies against substance P or somatostatin or with a mixture of both. Substance P-like immunoreactivity was found in 18.1% (untreated ganglia) and 19.6% (colchicine treated ganglia) of the labeled neurons. After incubation with a mixed antibody solution, 18.2% of joint afferents in untreated and 19.9% of the cells in colchicine-treated ganglia were immunopositive. Comparing this result with the results obtained using somatostatin and substance P antibodies alone, one can calculate that both neuropeptides are colocalized in about 17% of the cat's knee joint afferents. About 3% of the neurons contain only substance P, whereas almost none of the neurons contain only somatostatin. Based on this fact, one can assume that both neuropeptides are coreleased in peripheral tissue as well as in the central nervous system. © 1995 Wiley-Liss, Inc.     

Calbindin-immunoreactive sensory neurons of dorsal root ganglion project to skeletal muscle in the chick

In the chicken dorsal root ganglia, two neuronal subpopulations referred to as A1 and B1 share in common an immunoreactivity to antisera raised to calbindin D-28k but are distinguished by their cytological and ultrastructural characteristics. To determine the peripheral targets innervated by calbindin-immunoreactive neurons in lumbosacral dorsal root ganglia, cryostat sections of various hindlimb tissues were treated with anticalbindin antisera. Calbindin-immunostained axons were clearly detected in skeletal muscle. Large myelinated nerve fibres and afferent axon terminals in neuromuscular spindles were calbindin-immunoreactive; thin unmyelinated nerve fibres were also immunostained in nerve bundles of the perimysium. Since motoneurons and neurons of the autonomic nervous system were devoid of calbindin immunostaining, it was suggested that the immunoreactive axons found in skeletal muscle originate from sensory neurons expressing a calbindin immunoreaction in the dorsal root ganglia. This hypothesis was corroborated after introduction of wheat germ agglutinin coupled with horseradish peroxidase or colloidal gold particles into the sartorius muscle. The retrogradely transported tracer was collected only in ganglion cell bodies which displayed the ultrastructural characteristics of A1 and B1 sensory neurons. On the basis of calbindin immunoreaction and of tracer retrograde transport, it is concluded that ganglion cells of subclasses A1 and B1 contribute to the sensory innervation of skeletal muscle in the chicken.     

The role of neuropeptides in the sacral autonomic reflex pathways of the cat

Immunohistochemical and pharmacological studies were conducted to examine the origin and function of peptidergic nerves in the sacral autonomic system of the cat. Leucine-enkephalin (L-Enk) immunoreactivity was identified in nerve terminals in peripheral ganglia on the surface of the urinary bladder and in the parasympathetic nucleus in the sacral spinal cord. In colchicine-treated animals L-Enk was also detected in sacral preganglionic neurons (sPGN) identified by retrograde transport of a fluorescent dye. L-Enk terminals in bladder ganglia are believed to arise from sPGN since the terminals were eliminated by transection of the sacral ventral roots. Pharmacological studies indicated that exogenous as well as endogenously released enkephalins have an inhibitory action at both ganglionic and spinal sites in the sacral outflow to the urinary bladder. Peptides were also associated with afferents nerves in the sacral autonomic system. The distribution of substance P, VIP and cholecystokinin in the sacral dorsal horn paralleled the distribution of visceral afferent projections as demonstrated with HRP techniques. Dye labeling combined with immunohistochemistry revealed that some dorsal root ganglion cells projecting to the pelvic viscera contain substance P or VIP.     

Composition and central projections of the pudendal nerve in the rat investigated by combined peptide immunocytochemistry and retrograde fluorescent labelling

The central distribution of the afferent and efferent projections of pelvic striated muscles, the pudendal and sciatic nerves, were systemically investigated in rats by retrograde tracing techniques combined with immunocytochemistry using antibodies to 9 neuropeptides. True Blue was injected into either the pelvic muscles, pudendal or sciatic nerves. Seven days later the spinal cord and dorsal root ganglia (L3-S2 levels) were processed for immunocytochemistry. Injection of tracer into the pelvic muscles labelled dorsomedial, ventral and dorsolateral motoneuron groups of the L6 segment and a few sensory neurons in the respective dorsal root ganglia. Pudendal nerve injection also labelled the same motoneuron groups, 50% of neurons of the retrodorsolateral column and numerous sensory neurons of the dorsal root ganglion. Concomitant labelling of pudendal and sciatic nerves with different fluorescent tracers revealed a small number of double-labelled cells in the dorsal root ganglia but only single-labelled cells in the retrodorsolateral nucleus. Enkephalin-, somatostatin- and neuropeptide Y-containing fibres were particularly abundant in and around dorsomedial and dorsolateral groups as well as the intermediolateral cell column. We conclude that in the rat (a) the pudendal nerve has motor, sensory and autonomic (parasympathetic) components in contrast to the sciatic which is primarily motor and somatosensory, (b) some afferents from these nerves exhibit pre-spinal convergence and (c) dorsomedial and dorsolateral motoneuron groups are homologous to Onuf's nucleus in man.     

Differential vulnerability of neurochemically identified subpopulations of retinal neurons in a monkey model of glaucoma

The vulnerability of subpopulations of retinal neurons delineated by their content of cytoskeletal or calcium-binding proteins was evaluated in the retinas of cynomolgus monkeys in which glaucoma was produced with an argon laser. We quantitatively compared the number of neurons containing either neurofilament (NF) protein, parvalbumin, calbindin or calretinin immunoreactivity in central and peripheral portions of the nasal and temporal quadrants of the retina from glaucomatous and fellow non-glaucomatous eyes. There was no significant difference between the proportion of amacrine, horizontal and bipolar cells labeled with antibodies to the calcium-binding proteins comparing the two eyes. NF triplet immunoreactivity was present in a subpopulation of retinal ganglion cells, many of which, but not all, likely correspond to large ganglion cells that subserve the magnocellular visual pathway. Loss of NF protein-containing retinal ganglion cells was widespread throughout the central (59–77% loss) and peripheral (96–97%) nasal and temporal quadrants and was associated with the loss of NF-immunoreactive optic nerve fibers in the glaucomatous eyes. Comparison of counts of NF-immunoreactive neurons with total cell loss evaluated by Nissl staining indicated that NF protein-immunoreactive cells represent a large proportion of the cells that degenerate in the glaucomatous eyes, particularly in the peripheral regions of the retina. Such data may be useful in determining the cellular basis for sensitivity to this pathologic process and may also be helpful in the design of diagnostic tests that may be sensitive to the loss of the subset of NF-immunoreactive ganglion cells.     

Parkinson's disease with involvement of the parasympathetic ganglia

We report the distribution of Lewy bodies in an 83-year-old man who was diagnosed histopathologically as having Parkinson's disease. Many sections were taken from the brain, spinal cord, spinal dorsal root ganglia and peripheral autonomic systems. In the central nervous system, Lewy bodies were present in many of the areas already reported by previous authors. In the peripheral nervous system, Lewy bodies were present in the sympathetic ganglia, enteric nervous system of the alimentary tract and the submandibular ganglion, which is a peripheral parasympathetic ganglion. The present case indicates that the peripheral parasympathetic ganglia are involved in the disease process of Parkinson's disease.     

3,3′-Iminodipropionitrile induces neurofilament accumulations in the perikarya of rat vestibular ganglion neurons

Exposure of rats to 3,3′-iminodipropionitrile (IDPN) results in neurofilament (NF)-filled swellings in the proximal axons of a number of large neurons, including sensory neurons in the dorsal root ganglia (DRG) and motor neurons in the spinal cord. The present report describes the effects of acute and chronic IDPN exposure on the vestibular ganglion (VG) neurons as compared to those on the DRG neurons. In the VG, IDPN induced intra-perikaryal accumulation of morphologically and immunocytochemically identified NFs. In the DRG of the same treated animals, IDPN induced proximal axonal swelling but no perikaryal NF accumulations. We concluded that the VG neurons preferentially express the IDPN-induced NF pathology in their myelinated cell bodies. It is hypothesized that the NF pathology occurring after IDPN is preferentially expressed in myelinated structures.     

Parasympathetic innervation of cephalic arteries in rabbits: Comparison with sympathetic and sensory innervation

We investigated the distribution of parasympathetic, sympathetic, and sensory perivascular nerve fibers in rabbit cephalic arteries supplying the brain, exocrine glands, nasal mucosa, masseter muscles, tongue, and skin in the face and also examined cranial autonomic and sensory ganglia. NADPH diaphorase (NADPHd)-positive and vasoactive intestinal peptide–like immunoreactive (VIP-LI) neurons were located in the cranial parasympathetic ganglia. Neuropeptide Y (NPY)-LI neurons occurred mainly, and dopamine β-hydroxylase (DBH)-LI neurons occurred exclusively, in the superior cervical (sympathetic) ganglion. Substance P (SP)-LI and calcitonin gene-related peptide (CGRP)-LI neurons occurred only in the trigeminal (sensory) ganglion. Therefore, it was assumed that NADPHd-positive and VIP-LI perivascular nerve fibers in cephalic arteries were parasympathetic, all DBH-LI and most NPY-LI fibers were sympathetic, and SP-LI and CGRP-LI fibers were sensory in nature. In the cerebral arteries, NADPHd-positive and VIP-LI varicose fibers were more numerous in the rostral than in the caudal half of the Circle of Willis. In the extracranial arteries, NADPHd-positive and VIP-LI fibers were most abundant in the lingual, lacrimal, and supraorbital arteries; sparse in the parotid and submandibular arteries; and absent in the ear artery. There was an obvious proximal-to-distal density gradient along individual cephalic arterial trees. In contrast, DBH-LI, NPY-LI, SP-LI, and CGRP-LI varicose nerve fibers were similar in density in all cephalic arteries and their branches. These neuroanatomical findings suggest that differential parasympathetic innervation in cephalic arteries may play a role in the partitioning of blood flow between different cephalic tissues. J. Comp. Neurol. 389:484–495, 1997. © 1997 Wiley-Liss, Inc.     

Calretinin-containing neurons which co-express parvalbumin and calbindin D-28k in the rat spinal and cranial sensory ganglia; triple immunofluorescence study

The co-expression of calretinin with parvalbumin and calbindin D-28k was examined in the rat cranial and spinal sensory ganglia by triple immunofluorescence method. In the trigeminal and nodose ganglia, 9% and 5% of calretinin-immunoreactive neurons, respectively, also contained both parvalbumin- and calbindin D-28k immunoreactivity. These neurons had large cell bodies. In the trigeminal ganglion, they were restricted to the caudal portion. Such neurons were evenly distributed throughout the nodose ganglion. The co-expression could not be detected in the dorsal root, jugular or petrosal ganglia. Nerve fibers which co-expressed all the three calcium-binding proteins were observed in the inferior alveolar nerve but not the infraorbital nerve or palate. In the periodontal ligament, these nerve fibers formed Ruffini-like endings. These findings suggest that (1) the co-expression in trigeminal neurons is intimately related to their peripheral receptive fields; (2) the three calcium-binding proteins (calretinin, parvalbumin, calbindin D-28k) co-expressed in the trigeminal neurons may have mechanoreceptive function in the periodontal ligament.     

Neuropeptide proctolin (H-Arg-Try-Leu-Pro-Thr-OH): immunocytochemical mapping of neurons in the central nervous system of the cockroach

Proctolinlike immunoreactivity was mapped in the central nervous system of the cockroach, Periplaneta americana, by using whole-ganglion immunoreacted preparations. The procedure for this immunohistochemical staining of whole-mounts is described. Immunoreactivity was confined to neuronal cell bodies and and processes. These were found in all ganglia of the CNS. The cells varied in the consistency and intensity of their staining. The occurrence and variability of staining is described in detail. Cell bodies were found in the dorsal, ventral, and lateral regions of the ganglia. The highest number of cell bodies was found in the terminal ganglion and the lowest number in the cerebral ganglion. Those in the cerebral ganglion occurred mainly in the tritocerebral lobes. The distribution of immunoreactive cell bodies correlated with results previously obtained by radioimmunoassay. Immunoreactive processes were detected in all interganglionic connectives and many ganglionic nerve roots. Dense ramifications of immunoreactive processes and variocosities were detected in many of the ganglia. The widespread presence of immunoreactivity suggests that proctolin has diverse central and peripheral functions. The mapping immunoreactive neuronal somata provides a valuable step in the identification of putative proctolin-containing neurons suitable for further biochemical, anatomical, and physiological analysis.     

Galanin immunoreactivity in the mudpuppy cardiac ganglion.

The source of galanin-immunoreactive fibers in the cardiac ganglion and on cardiac muscle in mudpuppy (Necturus maculosus) has been determined utilizing immunohistochemical techniques. The galanin-immunoreactive fibers are not processes of afferent fibers originating in either the rostral four dorsal root ganglia or vagal sensory ganglia. Following colchicine treatment, all of the postganglionic parasympathetic neurons and a subpopulation of the small intrinsic neurons in the cardiac ganglion exhibit galanin immunoreactivity. The majority of the galanin-immunoreactive fibers that form complexes on the parasympathetic postganglionic neurons are derived from galanin-immunoreactive small intrinsic neurons, although some of these connections may represent collateral processes from other parasympathetic postganglionic neurons. All of the galanin-immunoreactive processes that innervate cardiac muscle are derived from postganglionic parasympathetic neurons in the cardiac ganglion.     

Galanin-like immunoreactivity in capsaicin sensitive sensory neurons and ganglia

In rats treated with capsaicin (CAP) as neonates, galanin-like (GA) immunoreactivity is markedly decreased in the trigeminal ganglion and the dorsal root ganglia as well as in the superficial layers of the dorsal spinal cord (laminae I and II), the substantia gelatinosa, the nucleus and tractus of the spinal trigeminal nerve and the nucleus commissuralis. Since CAP causes selective degeneration of primary sensory neurons of the C-fiber type and type B-cells of sensory ganglia, it is concluded that GA in CAP-sensitive primary sensory neurons represents a novel peptidergic system possibly involved in the transformation or modulation of peripheral nociceptive impulses. This system differs from the CAP-resistant GA-like neurons in other brain areas.     

Immunocytological localization of pedal peptide in the central nervous system and periphery of Aplysia

Immunocytology using antisera raised to conjugated pedal peptide (Pep) was used to localize the peptide in the CNS and periphery of Aplysia. A total of over 200 neurons in the CNS exhibited Pep-like immunoreactivity. As expected from results presented in the previous paper, immunoreactive neurons were heavily concentrated in the pedal ganglia, primarily in a broad ribbon comprised of about 60 large contiguous neurons on the dorsal side of each ganglion. Smaller and less numerous immunoreactive neurons were found in the other ganglia. A number of neurons primarily located in the abdominal ganglia had dense networks of immunoreactive varicose fibers surrounding their cell bodies. Many immunoreactive axons were observed in peripheral nerves, particularly those nerves leaving the pedal ganglia. Analyses of sections of body wall indicated that Pep-like immunoreactivity was localized to a series of varicose axons that appeared to be associated with vascular spaces, muscle fibers, and other large cells. These axons likely arise from pedal ganglion nerves that were shown to transport large amounts of 35S-labeled Pep to the periphery. These results suggest that Pep is a transmitter-like neuropeptide that is likely to have a number of important physiological actions in Aplysia.     

The origin of sensory nerve fibers that innervate the submandibular salivary gland in the rat

The origin of sensory nerves that innervate the submandibular salivary gland was investigated in the rat. After application of wheat germ agglutinin-horseradish peroxidase to the cut endings of the sympathetic and parasympathetic nerve branches at the hilus of the gland, labeled cells were mainly found in the dorsal root ganglia and the trigeminal ganglion, respectively. The labeled neurons in these ganglia were of various sizes compared to unlabeled neurons, suggesting that the sensory nerves of the gland conduct various modalities of sensory information.