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During rabies virus infections, the minor salivary glands are one of the important organs for virus replication and excretion into the oral cavity. However, details of pathological findings and viral antigen distribution in the minor salivary glands remain poorly understood. In this study, we conducted pathological tests on the tongues of 71 rabid dogs in the Philippines; the minor salivary glands (von Ebner's glands, lingual glands), circumvallate papilla, autonomic ganglia, and skeletal muscles were evaluated. Inflammatory changes were observed in the von Ebner's glands of 20/71 dogs, in the circumvallate papilla of 10/71, and in the tongue muscle of 1/71. Conversely, no morphological changes were observed in the lingual glands and autonomic ganglia. Viral antigens were detected via immunohistochemistry-based methods in the cytoplasm of the acinar epithelium in the von Ebner's glands of all 71 dogs. Virus particles were confirmed in the intercellular canaliculi and acinar lumen via electron microscopy. In the autonomic ganglia, viral antigens were detected in 67/71 rabid dogs. Viral antigens were detected in the taste buds of all 71 dogs, and were distributed mainly in type II and III taste bud cells. In tongue muscle fibers, viral antigens were detected in 11/71 dogs. No virus antigens were detected in lingual glands. These findings suggest that rabies virus descends in the tongue along the glossopharyngeal nerve after proliferation in the brain, and von Ebner's glands and taste buds are one of the portals of virus excretion into the saliva in rabid dogs.

Aging can promote significant morphofunctional changes in the gastrointestinal tract (GIT). Regulation of GIT motility is mainly controlled by the myenteric neurons of the enteric nervous system. Actions that aim at decreasing the aging effects in the GIT include those related to diet, with caloric restriction (CR). The CR is achieved by controlling the amount of food or by manipulating the components of the diet. Therefore, the objective of this study was to evaluate different levels of CR on the plasticity of nicotinamide adenine dinucleotide phosphate- (NADPH-) reactive myenteric neurons in the colon of Wistar rats during the aging process using ultrastructural (transmission electron microscopy) and morphoquantitative analysis. Wistar male rats (Rattus norvegicus) were distributed into 4 groups (n = 10/group): C, 6-month-old animals; SR, 18-month-old animals fed a normal diet; CRI, 18-month-old animals fed a 12% CR diet; CRII, 18-month-old animals fed a 31% CR diet. At 6 months of age, animals were transferred to the laboratory animal facility, where they remained until 18 months of age. Animals of the CRI and CRII groups were submitted to CR for 6 months. In the ultrastructural analysis, a disorganization of the periganglionar matrix with the aging was observed, and this characteristic was not observed in the animals that received hypocaloric diet. It was observed that the restriction of 12.5% and 31% of calories in the diet minimized the increase in density and cell profile of the reactive NADPH neurons, increased with age. This type of diet may be adapted against gastrointestinal disturbances that commonly affect aging individuals.

Backgound: The role of enteric nerves has previously been demonstrated in the formation of several gastric diseases. In the present review, the significance of the cholinergic nerves in stress-induced ulcer formation as well as the importance of substance P in the formation of gastric MALT lymphoma is discussed., Methods: The stress-induced ulcer was induced by the plaster bandage methods in rats. The gastric MALT lymphoma was formed by the peroral infection of gastric mucosal homogenate of the infected mouse in C57BL/6 mice. For the stress-induced ulcer, the distribution of the cholinergic nerves and muscarinic acetylcholine receptors was investigated by acetylcholinesterase histochemistry and autoradiography of water soluble compounds using 3H-quinuclidinyl benzilate was performed. To the MALT lymphoma study, the distribution of the substance P and effect of substance P antagonist, spantide II, was investigated by immunohistochemical studies., Results: The stress induced ulcer formation was shown to be related to the hyperactivity of the cholinergic nerves. The gastric MALT lymphoma was shown to be related to the increased localization of substance P., Conclusion: Stress-induced ulceration as a model of hyperactivity of the cholinergic nerves was proved to be a useful approach, while substance P and its role in MALT lymphoma formation may serve as a tool to clarify the neuroimmune modulation of chronic infectious diseases., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.)

From the perspective of neuropathies with an acute onset mimicking that of Guillain-Barré syndrome (GBS), cases with profound sensory and/or autonomic impairment without any significant weakness have been reported. Although the possibility of infectious or toxic etiologies should be carefully excluded, immune mechanisms similar to those in GBS are suggested to be involved in these so-called acute sensory neuropathies and acute autonomic neuropathies. The types of neuropathy include those with predominant sensory manifestations, predominant autonomic manifestations such as autoimmune autonomic ganglionopathy, and both sensory and autonomic manifestations such as acute autonomic and sensory neuropathy. Neuronopathy in the sensory and/or autonomic ganglia (i.e., ganglionopathy) has been commonly suggested in patients with these types of neuropathies. The presence of Anti-GD1b antibodies has been reported in some of the patients with acute sensory neuropathy with deep sensory impairment, whereas anti-ganglionic acetylcholine receptor antibodies are reported to be present in half of the patients with autoimmune autonomic ganglionopathy. The discovery of anti-ganglionic acetylcholine receptor antibodies significantly expanded the spectrum of autoimmune autonomic ganglionopathy. This is because some of the patients with chronic progression mimicking neurodegenerative diseases such as pure autonomic failure were positive for these antibodies. In contrast, pathologically significant autoantibodies have not been identified in acute autonomic and sensory neuropathy. Further studies are needed to clarify the pathogenesis and the spectrum of these types of neuropathies.

The neural apparatus and the endocrine part of the pancreas was studied in Wistar rats aged 3-4 and 19 months (n = 24) using the immunohistochemical reactions for synaptophysin (Syn), tyrosine hydroxylase (TH) and protein gene product 9.5 (PGP 9.5). Since Syn and PGP 9.5 are highly selective in detection of pancreatic islet (PI) endocrinocytes, it was possible to examine their topography and density in all parts of the pancreas. It was found that in rats aged 19 months, the total number of PI was decreased as compared to that in young animals. The study of PI size distribution has shown that the number of large islets decreased with age. Young animals showed rich innervation of the pancreas which was represented by three nerve plexuses: the first was a broadly-looped one, formed by small nerve trunks and bundles of unmyelinated and myelinated nerve fibers, the second consisted of thin bundles of postganglionic axons and microganglia, and the third (main terminal plexus) was formed by axons with varicosities and synapses of "en passant" type. In aged rats, marked degenerative changes in the neurons of intramural ganglia, nerve trunks and bundles were noted together with the reduction or complete absence of Syn- and TH-positive efferent parasympathetic and sympathetic terminals around blood vessels, excretory ducts, denervation of the exocrine and endocrine parts of the pancreas. Innervation disturbances in some lobules were accompanied by small inflammatory perivascular infiltrates.

This study shows for the first time the presence of intra and extrapancreatic ganglionar neurons and telocytes in Octodon degus such as those described in human and guinea pig pancreas. Pancreatic ganglionar neurons were identified by their histological characteristics as well as their positive immunostaining with mouse anti-human neuron specific enolase (NSE) antibody. Somatostatin secreting delta cells (D cells) in the islets of Langerhans were identified by positive immunostaining with rabbit antihuman polyclonal somatostatin antibody. Electron microscopy evidenced the presence of some unmyelinated axons in the interlobular spaces or septa, usually located adjacent to blood vessels and the exocrine epithelial ducts. The presence of telocytes with at least 2 telopodes was observed in the interlobular space, frequently in close spatial relationship with blood vessels and nerve endings. Telocytes were often observed in the vicinity or even in close proximity with both secretory acini and exocrine epithelial ducts and regulatory nerves and blood vessel apparatuses. A possible framework has been put forward within which such structures might contribute to elicit physiological responses in the pancreas. Further studies of synaptic interactions within and between pancreatic neuron cells are needed to help clarify the morphological results reported here. A broad overview of the field of neurogastroenterology with focus on the pancreas of O. degus related to the enteric nervous system (ENS) is provided in order to help design future studies on the connections of specific neurons forming pancreatic pathways, their neurotransmission processes and how disruption of these pathways may contribute to pancreatic disease., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Ganglionic synapses have the capability to express long-term potentiation (gLTP) after application of a brief high-frequency stimulus. It has been suggested a possible role of gLTP in some cardiovascular diseases. Although a number of characteristics of gLTP have been described, the precise locations and mechanisms underlying gLTP are not completely known. Current findings support two major conflicting presynaptic and postsynaptic hypotheses. The presynaptic hypothesis posits a presynaptic increase in acetylcholine (ACh) release, whereas the postsynaptic hypothesis proposes a long-lasting enhancement of the nicotinic response on the postsynaptic membrane. An alternative trans-synaptic hypothesis proposes the presynaptic release of a cotransmitter from large dense core vesicles, which postsynaptically enhances synaptic efficacy and accounts for gLTP. Here, we review the studies of LTP, with emphasis on gLTP in mammals, and we examine the findings that support the presynaptic, the postsynaptic and the trans-synaptic hypotheses. We then review our data on the contribution of calcium to gLTP as an approach to elucidate the mechanisms of gLTP. Data on the contribution of calcium to gLTP and on prolonged high-frequency stimulus-dependent fading of LTP have led us to support the trans-synaptic process as responsible for gLTP. Finally, we present a formal working model for the mechanisms of gLTP., (Copyright © 2013 Elsevier Inc. All rights reserved.)

A 14-year-old boy with mitochondrial neurogastrointestinal encephalopathy (MNGIE) disease had a lifelong history of failure to thrive and gastrointestinal symptoms including vomiting, pain, and diarrhea, leading to progressive cachexia. At the age of 9 years, after an extensive workup, the diagnosis of Crohn disease was strongly suspected, and he underwent colonoscopy with multiple biopsies. At 11 years of age, vision change and poor balance lead to a diagnosis of leukodystrophy by magnetic resonance imaging. Investigations for metachromatic leukodystrophy, adrenal leukodystrophy, and globoid cell leukodystrophy were all negative. A diagnosis of MNGIE disease was suspected when he continued deteriorating with gastrointestinal symptoms, multiple neurologic deficits, and encephalopathy. Markedly diminished thymidine phosphorylase activity and increased thymidine plasma levels confirmed the diagnosis of MNGIE. At autopsy, megamitochondria were observed by light microscopy in submucosal and myenteric ganglion cells and in smooth muscle cells of muscularis mucosae and muscularis propria, along the entire gastrointestinal tract from the esophagus to the rectum. Megamitochondria in ganglion cells were also observed in a retrospective review of the endoscopic intestinal biopsies taken at age 9 and 13 years and in the appendectomy specimen obtained 1 month before his demise. This study corroborates the presence of megamitochondria in gastrointestinal ganglion cells in MNGIE disease, better illustrates their detailed morphology, and describes for the first time similar structures in the cytoplasm of gastrointestinal smooth muscle cells. Pathologists should be able to recognize these structures by light microscopy and be aware of their association with primary mitochondriopathies., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Background: Pterygopalatine ganglion (PPG) branches, seem to be involved in the pathophysiology of facial pain. The functions of these branches, including a recently discovered orbital branch, are not completely known but could be of clinical significance. This study was designed to characterize PPG branches through immunohistochemical stain and study their anatomy, specifically the orbital branches., Methods: In a cadaver study of four specimens, the pterygopalatine fossa (PPF) was dissected out of its bony surroundings as a tissue block. Subsequently, cryostat sectioning of these blocks was performed. In one specimen the PPF was microscopically dissected. Recently discovered neural structures were identified, dissected out of the tissue block, and cryosectioned. All cryostat sectionings were immunohistochemically stained for protein gene product (PGP) 9.5, nitric oxide synthase (NOS), and tyrosine hydroxylase (TH)., Results: A recently discovered neural connection between the PPG and the ophthalmic nerve could be confirmed and classified as an orbital PPG branch. The connection stained throughout for PGP 9.5 and partially stained for NOS. In other orbital branches, both NOS and TH(+) nerve fibers were found. The PPG contained NOS(+) cells. TH labeling was also found in nerve fibers running through the PPG and the vidian nerve., Conclusion: The recently discovered orbital PPG branch is of a mixed parasympathetic and sensory nature. In the other orbital branches, sympathetic fibers were shown as well. This knowledge may add to understanding the symptomatology and therapies of headache syndromes such as nerve block.

Objective: To study the aging of submucous plexus of the small intestine (jejunum-ileum) of the guinea pigs from the quantitative, structural and ultrastructural perspective., Method: Chemical preparations of membrane of the jejunum-ileum of old and young animals with the use of light and electronic microscope., Results: The ganglia of young animals presented between 1 and 56 neurons and the old animals presented from 1 to 30 neurons. The mean density of the ganglia by cm(2) in the young jejunum-ileum was of 551±36.89 and in the old one 413±11.86. The density of the neurons was 5011±291.11 neurons/cm(2) average in young animals and 2918±120.70 neurons/cm(2) in the old ones. The size of the neurons varied in both age groups. The collagen fibers in the ganglia of old animals they were condensed. Degenerated mitochondrias in the interior of the cell were frequent in the old animals., Conclusion: In submucous plexus of the jejunum-ileum there is a loss of 38% of the neurons with aging.

Using the immunohistochemical methods for detection of neurofilaments (NF), peripherin (PRF), synaptophysin (SF), tyrosine hydroxylase (TH) and Nissl-staining with toluidine blue, thick sections (10-20 microm) prepared through all parts of pancreas of adult Wistar rats (n=7) were studied. The topography and the density of distribution of pancreatic islets in the various parts of pancreas were defined. The greatest density of the islets was found in the body of pancreas. The dense innervation of the organ was detected which included several nervous plexuses: the big-looped one consisting of nervous bundles and small trunks of NF-positive myelinated and unmyelinated nervous fibers, the second one was formed by PRF-positive thin posganglionic bundles of axons and microganglia, and the third one, detected with SF-staining, was main terminal plexus consisting of varicose axons with en passant synapses. Interactions of synaptophysin-positive terminals (distant en passant synapses) with blood vessels, endocrine (islet) and exocrine cells, excretory ducts of the pancreatic lobules are described in details. Peculiarities of the structure of parasympathetic ganglia, their neurons, and pericellular synaptic apparatus are described; problems of the innervation and the nature of pancreatic insular endocrine cells are discussed. Attention is drawn to the fact that in no case neurons were found in the islets in the rat pancreas.

The fact that most published data on syncytial cytoplasmic anastomoses are based on the autonomic nervous system in the early postnatal period of development, when many nerve fibers are poorly ensheathed by glia or have no glial sheaths at all, has led to the assumption that these anastomoses do not exist in adults because of the significant development of the glia and glial insulation of individual neurites from each other. We tested this assumption using electron microscopic studies of the caudal mesenteric ganglion in adult cats. A high level of glial ensheathing of neurites was observed. However, syncytial pores were seen between contacting neurites lacking glial sheaths in almost every specimen. This is the first report describing axodendritic synapses with perforations in the presynaptic zone outside the synaptic specializations in the autonomic nervous system. It is suggested that although syncytial cytoplasmic connections are seen in adult animals, they do not contradict the neuron theory.

Fragile X tremor/ataxia syndrome (FXTAS) occurs in individuals with moderate CGG expansion of the fragile X mental retardation 1 (FMR1) gene and is associated with intranuclear inclusions in neurons and astrocytes. Although the neuropathologic findings in the brain and spinal cord were described, pathological features in the peripheral nervous system were not reported. Here, we report on novel neuropathological findings in the peripheral nervous system and especially in autonomic ganglia at autopsy in a man with FXTAS. In addition to the characteristic brain and spinal cord findings, typical intranuclear inclusions were identified in the ganglion cells of adrenal medulla, dorsal root ganglia, paraspinal sympathetic ganglia, myenteric ganglia of the stomach and subepicardial autonomic ganglion of the heart. Our findings indicate that FXTAS diffusely involves the central and peripheral nervous systems, which explains the protean neurological symptoms ranging from dementia to dysautonomia.

The membranes of neuron profiles adjacent to each other in the caudal mesenteric ganglion were examined electron microscopically during the first 3 days after crossing of the preganglionar or postganglionar branches. The contacting membranes were thinned and perforated, and neuron-to-neuron syncytial connections were forming. Multiple connections were formed between the nerve processes and terminals in the synapse. The pre- and even postsynapses were clear in pronounced perforations.

Synaptic activity is thought to be critical for synaptic stabilization. In this issue of Neuron, Krishnaswamy and Cooper show that nicotinic synapses on autonomic neurons remain intact without synaptic activity. Postsynaptic responses are required, however, for presynaptic terminals to acquire the high-affinity choline transporter necessary for high-frequency transmission.

Since the majority of the published data on syncytial cytoplasmic anastomoses relate to the autonomic nervous system in the early postnatal period of development, when many nerve fibers are still poorly covered by glia or have no glial sheaths at all, it was suggested that such anastomoses were not present in adults due to the significant development of glia separating individual neurites from each other. To check this assumption, we have performed an electron microscopic study of the adult cat dorsal caudal mesenteric ganglion. The cell neurites were found to be frequently covered by glial sheaths. However, almost in every sample, the syncytial pores were detected between the contacting neurites lacking glial covering layers. Sometimes serial syncytially connected neurites were seen. Axo-dendritic synapses with presynaptic perforations outside the synaptic specializations were described in the autonomic nervous system for the first time. These observations therefore provide evidence of syncytial cytoplasm connections in normal adult animals, however this does not reject the neuronal doctrine.

Final motor neurons in sympathetic and parasympathetic ganglia receive synaptic inputs from preganglionic neurons. Quantitative ultrastructural analyses have shown that the spatial distribution of these synapses is mostly sparse and random. Typically, only about 1%-2% of the neuronal surface is covered with synapses, with the rest of the neuronal surface being closely enclosed by Schwann cell processes. The number of synaptic inputs is correlated with the dendritic complexity of the target neuron, and the total number of synaptic contacts is related to the surface area of the post-synaptic neuron. Overall, most neurons receive fewer than 150 synaptic contacts, with individual preganglionic inputs providing between 10 and 50 synaptic contacts. This variation is probably one determinant of synaptic strength in autonomic ganglia. Many neurons in prevertebral sympathetic ganglia receive additional convergent synaptic inputs from intestinofugal neurons located in the enteric plexuses. The neurons support these additional inputs via larger dendritic arborisations together with a higher overall synaptic density. There is considerable neurochemical heterogeneity in presynaptic boutons. Some synapses apparently lack most of the proteins normally required for fast transmitter release and probably do not take part in conventional ganglionic transmission. Furthermore, most preganglionic boutons in the ganglionic neuropil do not form direct synaptic contacts with any neurons. Nevertheless, these boutons may well contribute to slow transmission processes that need not require conventional synaptic structures.

The accepted theory of vesicular release of neurotransmitter posits that only a single vesicle per synapse can fuse with the membrane following action potential invasion, and this exocytotic event is limited to the ultrastructurally defined presynaptic active zone. Neither of these dictums is universally true. At certain synapses, more than a single vesicle can be released per action potential, and there is growing evidence that neuronal exocytosis can occur from sites that are unremarkable in electron micrographs. The first discrepancy extends the dynamic range of synapses, whereas the second enables faster and more robust chemical transmission at sites distant from morphologically defined synapses. Taken together, these attributes expand the capabilities of cellular communication in the nervous system.

Intraganglionic laminar endings (IGLEs) represent the only vagal mechanosensory terminals in the tunica muscularis of the esophagus. Two specific markers for IGLEs were recently described in mouse: the purinergic P2 x 2 receptor and the vesicular glutamate transporter 2 (VGLUT2). This study aimed at comparing both markers with respect to their suitability for quantitative analysis. We counted IGLEs immunostained for VGLUT2 and P2 x 2, respectively, and mapped their distribution in esophageal wholemounts of C57Bl/6 mice. Numbers and distribution of IGLEs were compared with those of myenteric ganglia as demonstrated by cuprolinic blue histochemistry. Whereas the distribution of VGLUT2-immunopositive IGLEs closely matched that of myenteric ganglia, P2 x 2-immunopositive IGLEs were rarely found in upper and middle esophagus but increasingly in its lower parts. P2 x 2 stained only half the number of IGLEs found with VGLUT2 immunostaining. We also investigated the correlation between anterograde tracing and immunohistochemistry for identifying IGLEs. Confocal microscopy revealed colocalization of all three markers in approximately 50% of IGLEs. The remaining IGLEs showed only tracer and VGLUT2 labeling but no P2 x 2 immunoreactivity. Thus, VGLUT2 and P2 x 2 represent two specific markers for qualitative demonstration of esophageal IGLEs. However, VGLUT2 may be superior to P2 x 2 as a quantitative marker for IGLEs in the esophagus of C57Bl/6 mice.

Cardiac ganglia develop destructive ganglionitis in chronic Chagas' disease and rheumatic heart disease. This ganglionitis is associated with periganglionic infiltrations and is suspected of developing secondary to epicardial inflammation. If so, it would be expected that cardiac ganglia (1) are equipped with an inventory of immune competent cells allowing the initiation of inflammatory processes, and (2) are not effectively protected from the milieu of the surrounding tissue by metabolically active diffusion barriers. These problems were addressed in specified pathogen-free rats by electron microscopy and immunohistochemistry with markers for dendritic cells, monocytes/macrophages, and perineurial barriers. In contrast to nerve fascicles, cardiac ganglia are only partially enveloped by perineurial cells. Inside the ganglia, ramified cells with major histocompatibility complex class II antigen (reacting with monoclonal antibody OX6) on their surface and exhibiting an ultrastructure typical of dendritic cells are numerous, comprising nearly 5% of all cells within ganglia. The ratio of the number of these cells to that of neurons is 1:2. Cells reacting with monoclonal antibodies ED1 and ED2, markers for monocytes/macrophages, constitute 1.8% and 1.6% of the ganglionic cell population, respectively. Such cells are less frequent in the cervical trunk of the vagus nerve. Thus, the inventory of immune competent cells in rat cardiac ganglia is consistent with the view that the abundance of antigen-presenting cells correlates with the permeability of the barriers providing protection from blood-borne and tissue-borne factors.

The aim of the present study was to describe in detail the ultrastructure of intrinsic cardiac ganglionic cells in the healthy human as these cells appear to be directly involved in the development of tachycardia, atrioventricular block, ventricular fibrillation, and sudden cardiac death. Tissues examined in this study were obtained from hearts of 10 adult humans of either sex aged 22-80 years at autopsy performed no more than 8 h after death. The examined human intrinsic cardiac nerve cells were in most respects typical autonomic neurons surrounded by a sheath of satellite cells that was either uni- or multilayered. In addition to regular unmyelinated axons, prominent large axon terminals containing lamellated dense bodies, mitochondria and vesicles in the cytoplasm were observed in the ganglion neuropil. Synaptic profiles were more common in the ganglion neuropil than on neuronal somata. According to axon terminal contents, synaptic profiles were of three types. The most common Type 1 synaptic profiles contained a predominance of small clear, with a few larger dense-cored vesicles and mitochondria. Type 2 synaptic profiles, in addition to the same components as in Type 1, had glycogen-like particles. Type 3 vesicle-containing profiles clearly differed from both the previous ones as they were the largest in diameter and included plentifiul large clear pleomorphic or dense-cored vesicles together with small clear and larger dense-cored vesicles, mitochondria, dense and multivesicular bodies. Independently of age of the human, the most frequent neuronal abnormality was an abundant accumulation of inclusions inside of somata and dendrites that, in profile, appeared like circular membranous or fine granular bodies variable in electron density. In addition to inclusions, some neuronal somata and dendrites had strongly swollen mitochondria filled up with granular material in spite of their close association with normal looking ganglionic neurons. Structures resembling an axon growth cone in profile were revealed inside of cardiac ganglia derived from an 80 year old man. In conclusion, the present results provide baseline information on the normal ultrastructure of intracardiac ganglia in healthy humans which may be useful for assessing and interpreting the degree of damage of ganglionic cells both in autonomic and sensory neuropathies of the human heart.

Using a sensitive immunohistochemical technique, the localization of neuropeptide Y (NPY) Y1-receptor (Y1R)-like immunoreactivity (LI) was studied in various peripheral tissues of rat. Wild-type (WT) and Y1R-knockout (KO) mice were also analyzed. Y1R-LI was found in small arteries and arterioles in many tissues, with particularly high levels in the thyroid and parathyroid glands. In the thyroid gland, Y1R-LI was seen in blood vessel walls lacking alpha-smooth muscle actin, i.e., perhaps in endothelial cells of capillaries. Larger arteries lacked detectable Y1R-LI. A distinct Y1R-immunoreactive (IR) reticulum was seen in the WT mouse spleen, but not in Y1R-KO mouse or rat. In the gastrointestinal tract, Y1R-positive neurons were observed in the myenteric plexus, and a few enteroendocrine cells were Y1R-IR. Some cells in islets of Langerhans in the pancreas were Y1R-positive, and double immunostaining showed coexistence with somatostatin in D-cells. In the urogenital tract, Y1R-LI was observed in the collecting tubule cells of the renal papillae and in some epithelial cells of the seminal vesicle. Some chromaffin cells of adrenal medulla were positive for Y1R. The problem of the specificity of the Y1R-LI is evaluated using adsorption tests as well as comparisons among rat, WT mouse, and mouse with deleted Y1R. Our findings support many earlier studies based on other methodologies, showing that Y1Rs on smooth muscle cells of blood vessels mediate NPY-induced vasoconstriction in various organs. In addition, Y1Rs in other cells in parenchymal tissues of several organs suggest nonvascular effects of NPY via the Y1R., (Copyright 2002 Wiley-Liss, Inc.)

The neuroinsular complex type 1 is composed of pancreatic endocrine islet cells and nerve cell bodies intrinsic to the islet. The details of the relation between nerve cells and between endocrine cells and nerve cells in the complex are unknown. Pancreata from newborn and 18-day-old mice were analysed by electron microscopy to establish the ultrastructural morphology of the neuroinsular complex. Immunohistochemical staining for protein gene-product 9.5 was also performed. The study showed that nerve cell bodies were closely associated to each other in the periphery of the islets with no connective tissue separating the cells. The nerve cells were closely associated to both beta-cells and alpha-cells. Direct intercellular contacts were observed between nerve cells and endocrine cells and between Schwann cells and endocrine cells. Varicose nerve endings were frequently observed in the neuroinsular complex. In the peripheral parts the varicosities were mostly being associated to the nerve cell bodies. The varicosities contained small clear or small clear and larger dense cored vesicles, suggesting cholinergic and peptidergic contents. The varicosities made specialized synaptic connections with adjacently located nerve cells. The study shows that the neuronal part of the neuroinsular complex is closely associated to the endocrine islet cells and that it is richly innervated, indicating an important regulatory function of the nerve cell component in the neuroinsular complex.

Acetylcholine (ACh) stimulates contraction of the uterus and dilates the uterine arterial supply. Uterine cholinergic nerves arise from the paracervical ganglia and were, in the past, characterized based on acetylcholinesterase (AChE) histochemistry. However, the histochemical reaction for acetylcholinesterase provides only indirect evidence of acetylcholine location and is a nonspecific marker for cholinergic nerves. The present study: (1) reevaluated cholinergic neurons of the paracervical ganglia, (2) examined the cholinergic innervation of the uterus by using retrograde axonal tracing and antibodies against molecules specific to cholinergic neurons, choline acetyltransferase and the vesicular acetylcholine transporter, and (3) examined muscarinic receptors in the paracervical ganglia using autoradiography and a radiolabeled agonist. Most ganglionic neurons were choline acetyltransferase- and vesicular acetylcholine transporter-immunoreactive and were apposed by choline acetyltransferase/vesicular acetylcholine transporter-immunoreactive terminals. Retrograde tracing showed that some cholinergic neurons projected axons to the uterus. These nerves formed moderately dense plexuses in the myometrium, cervical smooth muscle and microarterial system of the uterine horns and cervix. Finally, the paracervical ganglia contain muscarinic receptors. These results clearly reveal the cholinergic innervation of the uterus and cervix, a source of these nerves, and demonstrate the muscarinic receptor content of the paracervical ganglia. Cholinergic nerves could play significant roles in the control of uterine myometrium and vasculature.

Although the pathogenesis of Hirschsprung's disease (HD) is not completely resolved, both the absence of nerve cells and the hypertrophy of nerve fascicles within the aganglionic colonic segment have been attributed to an abnormal intestinal microenvironment. Studies on animal models for HD revealed an altered ultrastructure of ingrowing nerve fascicles and abnormalities of basal laminae (BL). Therefore, the purpose of this study was to examine the ultrastructure of hypertrophied nerve fascicles in human HD with special reference to structural abnormalities of BL. Colonic specimens were obtained from patients with HD (n = 10) and controls (n = 5) and processed for electron-microscopical examination. Hypertrophied nerve fascicles were characterized by a prominent perineural sheath surrounded by large amounts of collagen bundles, a collagen-filled endoneurium, vasa nervorum and abundant glial cells of extraenteric ultrastructure, which were arranged in mono- or oligoaxonal units and frequently displayed different stages of myelination. As these ultrastructural characteristics resembled typical features of extrinsic nerves and were similar to those observed in subserosal nerves, the prominent intramural nerve fascicles were considered to be of extraenteric origin. Most likely their overabundance contributes to the functional obstruction of the terminal colon. Morphological abnormalities of BL encountered in the aganglionic colonic segment consisted of an extensive multilamination of BL surrounding glial processes and an irregular thickening of BL surrounding perineurocytes and smooth muscle cells of the muscularis mucosae. Similar alterations in BL have also been described in inherited peripheral and diabetic autonomic neuropathies and attributed to reactivated schwann cells. Thus, the overproduction of BL material within the hypertrophied nerve fascicles in HD may reflect an increased activity of proliferating glial cells. Since the smooth muscle cells of the muscularis mucosae showed abnormalities of BL similar to those observed in murine models for HD, it is suggestive that also in human HD the aganglionic colon is affected by a disturbed intestinal micro-environment impairing the neuronal colonisation and promoting the ingrowth of extrinsic nerves. The ultrastructurally observed alterations in BL of both neuronal and non-neuronal cells, as well as the increased amount of perineural and endoneural collagen provide further evidence that extracellular matrix components are abnormally distributed and overproduced within the bowel wall of patients affected by HD.

Neurocalcin is a newly identified neuronal calcium-binding protein. We tried here to investigate the immunohistochemical distribution of neurocalcin in the rat esophagus. Nerve cell bodies having neurocalcin immunoreactivity were found throughout the myenteric plexus. In the myenteric ganglia, two types of nerve terminals showed neurocalcin immunoreactivity. One was varicose terminals containing numerous small clear vesicles and forming a synapse with nerve cells. The other terminals were characterized by laminar or pleomorphic structure and many mitochondria. These laminar terminals were supposed to be sensory receptors of the esophageal wall. In the motor endplates of the striated muscles, nerve terminals containing many small clear vesicles and mitochondria also had neurocalcin immunoreactivity. After left vagus nerve cutting under the nodose ganglia, the number of immunopositive thick nerve fibers, laminar endings and nerve terminals on the striated muscles decreased markedly. Retrograde tracing experiments using Fast Blue showed extrinsic innervation of esophagus from ambiguus nucleus, dorsal motor nucleus of vagus, superior cervical ganglia, celiac ganglia, nodose ganglia and dorsal root ganglia. In the celiac ganglia, nodose ganglia and dorsal root ganglia, retrogradely labeled nerve cells were neurocalcin-immunoreactive. Neurons in the celiac ganglia may project varicose terminals, while nodose and dorsal root neurons project laminar terminals. Although cell bodies of motoneurons in the ambiguus nucleus lacked neurocalcin immunoreactivity, these neurons may contain neurocalcin only in the nerve terminals in the motor endplates. Neurocalcin immunoreactivity is distributed in many extrinsic and intrinsic neurons in the esophagus and this protein may play important roles in regulating calcium signaling in the neurons.

We examined by immunohistochemistry the expression of glial fibrillary acidic protein (GFAP) in enteric ganglia of the chick embryo, using a polyclonal antibody. The morphology of enteric ganglion cells was examined by electron microscopy. Faint GFAP immunoreactivity was detected in ganglion cells and cell processes from around day 7 in ovo. Later in development the intensity of the immunofluorescence increased and it became more evident that immunoreactive small ganglion cells (interpreted as primitive glial cells), and their processes, surrounded larger negative cell profiles (interpreted as primitive neuronal cells); GFAP immunofluorescence was also evident in intramuscular and mucosal nerve trunks. In colocalization experiments, GFAP immunoreactivity was detected in a proportion of HNK-1/N-CAM immunoreactive ganglion cells, in both the myenteric and submucosal plexus. In addition, we observed GFAP immunoreactive nerves in wholemount preparations of chick gut from as early as day 4.5 in ovo. In the ganglionated nerve of Remak, GFAP immunoreactive satellite and Schwann cells were in evidence from day 5 of incubation. Neuronal markers, such as neurofilament, have been detected very early in development in neural crest cell populations in chick enteric ganglia. In contrast, the expression of markers of the glial phenotype has previously been observed only in the late stages of embryonic development. From our experiments, we conclude that neuronal and glial phenotypes are immunohistochemically distinct from as early as day 4.5 of incubation, even if by ultrastructural criteria glial cells are clearly distinguishable from neurons only after day 16 in ovo., (Copyright 1998 Elsevier Science B.V.)

All nerve components of the sinus node (SN), adjacent working myocardium and nervous ganglions of 12 patients, from 9 to 50 years old, with an idiopathic syndrome of a long Q-T interval were studied electron-microscopically. Quantitative evaluation of the myocardium innervation density in this heart region was performed. Destructive changes of the myelinated and non-myelinated nervous fibers and gliocytes in the conductive myocardium of SN and working myocardium as well as in the neurocytes of the adjacent nervous ganglions were observed. Possible causes of the ultrastructural pathology in the heart sinoauricular region are discussed.

The innervation of the knee joint synovial membrane of the guinea pig, i.e., the synoviocyte layer, the subjacent connective tissue and the connective tissue region beneath, was analyzed with immunohistofluorescence and electron microscopy. A screening of the innervation with antibodies against the general axon marker -- protein gene product (PGP) 9,5 -- revealed the presence of nerve fibers distributed in various regions of the knee joint synovial membrane. Confirming previous studies, some of these nerve fibers stained with antibodies to tyrosine hydroxylase (TH), neuropeptide Y (NPY), substance P (SP), calcitonin gene-related peptide (CGRP), and vasoactive intestinal polypeptide (VIP). In addition, dynorphin (DYN)-containing fibers were detected, which have not been reported previously in normal joints. In general, the immunoreactive fibers were observed close to the synoviocytes and at blood vessels. Fibers with colocalization of NPY- and TH-like immunoreactivities (LIs), as well as of DYN- and TH-LIs were demonstrated. In the electron microscope, bundles of unmyelinated fibers as well as single fibers were found in the connective tissue region below the synoviocytes. Varicose parts of the nerve fibers contained mainly small, clear vesicles. Small and large dense-cored vesicles were also seen, but less frequently. Denser portions of the plasma membranes of some axons were observed in these regions, facing the extracellular space. Myelinated fibers were also observed in some nerve bundles. These findings emphasize the complex innervation of the synovial membrane, with nerve fibers containing a host of neuroactive substances. Altogether, these fibers are probably involved in many functions such as vasoregulation and control of synovial secretion in addition to being a source of mediators in joint inflammation.

In this 8-year-old boy, who had been exposed to alcohol and oxazepam during pregnancy, visual failure was the first symptom of a neuronal ceroid lipofuscinosis (NCL) disorder, noticed at the age of 5 years. Ophthalmological examinations revealed a cystic type of macular degeneration, which would be more likely to be found in variant late infantile NCL. However, vacuolated lymphocytes were found in peripheral blood films and a diagnosis of the juvenile form of NCL (JNCL) was made. Molecular genetic studies showed the patient to be homozygous for the major mutation of JNCL, a 1.02-kb deletion. In whole-night polysomnography, there was significantly more epileptiform activity than in other JNCL patients under 10 years of age. Using magnetic resonance imaging, the signal intensity of the white matter was increased, especially in the periventricular area. In addition, there were enlarged perivascular spaces in the watershead areas. The corpus callosum was thin. Finally, in the autonomic ganglion cells of the submucosal nerve plexus there were membrane-enclosed homogeneous and granular cytosomes resembling the granular osmiophilic deposits of infantile NCL. However, extraneural cells, including blood capillaries and smooth muscle, showed inclusions with fingerprint and curvilinear profiles. The features of the present case indicated a phenotypic variant of JNCL.

Spinal primary afferent fibres innervating the myenteric area in the oesophago-gastric junction of the cat were selectively labelled by anterogradely transported cholera toxin B subunit-horseradish peroxidase conjugate injected into thoracic dorsal root ganglia. The ultrastructure of these labelled primary afferent fibres was studied in order to determine whether they display close relationships with specific cell types in the myenteric plexus. Horseradish peroxidase was revealed with tetramethylbenzidine stabilized with ammonium heptamolybdate or with the tetramethylbenzidine/tungstate reaction in order to visualize the cytoplasmic organelles and the axolemma, respectively. The labelled primary afferent fibres were unmyelinated. Two kinds of profiles of labelled fibres containing vesicles and mitochondrial accumulations were found: (i) fibres running in myenteric connectives in isolated nerve bundles, and (ii) fibres within the myenteric ganglia. The first kind had small areas of axolemma with no glial cell covering, whereas the second kind had little or no glial cell covering (termed naked primary afferent fibres). In addition, labelled fibres containing few vesicles and mitochondria and running in nerve bundles surrounded by perineurium were numerous. Within the myenteric ganglia, naked primary afferent fibres contacted myenteric neurons. The contacts were mainly axosomatic. No synaptic specializations were distinguished. In the interganglionic area, some labelled fibres terminated close to blood vessels. The intraganglionic naked primary afferent fibres are suggested to be mechanoreceptors. Their exposed axolemma might allow both mechanotransduction and release of neurotransmitters which could act on myenteric neurons. Because they are protected by their glial cell sheath and by bundles of collagen fibrils, interganglionic primary afferent fibres are likely to be less exposed to deformation.

Peripheral innervation of the mouse pancreas was studied by scanning and transmission electron microscopy, as well as by light microscopy (cholinesterase technique). Major nerve bundles usually ran with arteries in the connective tissue septa. They gave off delicate branches that formed plexuses around arteries and arterioles. When reaching the capillaries, nerve fibers left the arterioles and formed very loose networks in the interacinar spaces. The nerves accompanying the arteries also sent off branches toward the islets of Langerhans and formed a dense plexus around the islets. A few delicate nerve fibers were also present around the pancreatic ducts. Thus, the intrapancreatic nerves formed four plexuses: perivascular, periductal, periacinar and peri-insular. The plexuses were networks of unmyelinated nerve fibers consisting of axons with varicosities and Schwann cells. Intrapancreatic ganglia were found in the interlobular connective tissue; ganglia were often closely associated to islets of Langerhans. Our findings indicate that the "interstitial cells" described by light microscopists correspond to Schwann cells. Axons in the nerve plexuses contain transmitter vesicles and therefore represent an autonomic terminal apparatus. The rich innervation of arterioles and islets suggests that neural regulation of secretory function is mediated by control of pancreatic blood flow.

Ultrastructural and immunocytochemical results of studies on interneuronal relationship in vegetative ganglia were represented. Age-related changes of synapses (appearance of synaptic type vesicles and smooth endoplasmic reticulum cisterns in postsynaptic terminals), peculiarities of interneuronal links in neurons containing neuropeptide Y and NO-synthase and structural manifestations of non-synaptic interneuronal links were shown. Modern views on interneuronal communications were discussed.

Tyrosine hydroxylase immunoreactive sensory neurons in the petrosal ganglion selectively innervate the carotid body via the carotid sinus nerve. Central projections of the carotid sinus nerve were traced with horseradish peroxidase. The commissural nucleus of the tractus solitarius was examined by dual labelling light and electron microscopy. Dense bilateral labelling with horseradish peroxidase was found in the tractus solitarius and commissural nucleus of the tractus solitarius. Horseradish peroxidase was found in unmyelinated axons, myelinated axons, and nerve terminals. About 88% of horseradish peroxidase-labelled carotid sinus nerve axons were unmyelinated. Tyrosine hydroxylase immunoreactivity was identified in unmyelinated axons, myelinated axons, dendrites, perikarya, and nerve terminals. Most tyrosine hydroxylase immunoreactive axons (93%) in the commissural nucleus of the tractus solitarius were unmyelinated. Tyrosine hydroxylase immunoreactivity was simultaneously identified in carotid sinus nerve unmyelinated axons, myelinated axons, and nerve terminals. These double-labelled terminals comprised 28% of the number of tyrosine hydroxylase immunoreactive terminals in the commissural nucleus of the tractus solitarius, and 55% of transganglionically-labelled terminals. Therefore, there are both central and peripheral sources of tyrosine hydroxylase immunoreactive nerve terminals in the commissural nucleus of the tractus solitarius. These data support the hypothesis that peripheral tyrosine hydroxylase immunoreactive neurons are involved in the origination of the chemoreceptor reflex. Axo-axonic synapses between peripheral carotid sinus nerve afferent terminals and central terminals containing tyrosine hydroxylase immunoreactivity were observed in 22% of the axo-axonic synapses observed. Thus, central tyrosine hydroxylase immunoreactivity neurons are involved in the modulation of the chemo-and/or baroreceptor reflexes. Synaptic contacts were not observed between carotid sinus nerve afferents and tyrosine hydroxylase immunoreactive perikarya of dendrites. Catecholaminergic neurons are thus unlikely to be the second order neurons of either the chemo-or baroreceptor reflex in the commissural nucleus of the tractus solitarius.

Findings of light optical and electron microscopy studies on formation of multinuclear neurons in gastrointestinal tract myenteric plexus in mammals affected by extreme factors (vagotomy, modelling of portal hypertension. Hyperbaric oxygenation and hypergravitation) were presented. A model for original transformation of multinuclear neuron as a mechanism of new nerve cells formation in the course of the organism compensatory-adaptive reaction to extreme action was obtained.

Exocrine secretion of the avian salt gland is controlled by the autonomic nervous system. NADPH-diaphorase histochemistry was employed at the light and electron microscopic level to provide the morphological basis for a putative nitrergic regulation of salt gland function. NADPH-diaphorase staining was localized in two cell populations of the parasympathetic secretory ganglion at high cell density and equal distribution throughout the ganglionic mass. In addition, salt gland-intrinsic neurons, arranged in small clusters and associated with major nitrergic fiber bundles, proved to be NADPH-diaphorase positive. These postganglionic nerve fibers innervated the secretory parenchyma in close proximity to the basal membrane of single secretory tubules as well as arterioles. The findings suggest participation of the nitrergic pathway in the autonomic control of avian salt gland function.

In the submucous plexus of the guinea-pig ileum, previous light-microscopic studies have revealed that vasoactive intestinal peptide (VIP)-immunoreactive and nitric oxide synthase (NOS)-immunoreactive terminals are found predominantly in association with VIP-immunoreactive nerve cell bodies. In this study, double-label immunohistochemistry at the light-microscopic level demonstrated co-localization of NOS-immunoreactivity and VIP-immunoreactivity in axon terminals in submucous ganglia. About 90% of nerve fibres with NOS-immunoreactivity or VIP-immunoreactivity were immunoreactive for both antigens; only about 10% of labelled varicosities contained only NOS-immunoreactivity or VIP-immunoreactivity. The VIP/NOS varicosities were more often seen in the central parts of the ganglia, close to the VIP-immunoreactive cell bodies. Ultrastructural immunocytochemistry with antibodies to VIP was used to determine if NOS/VIP terminals synapse exclusively with VIP-immunoreactive nerve cell bodies. We examined the targets of VIP-immunoreactive boutons in two submucous ganglia from different animals. Serial ultrathin sections were taken through the ganglia after they had been processed for VIP immunocytochemistry. For each cell body, the number of VIP inputs (synapses and close contacts) was determined. The number of VIP-immunoreactive synapses received by the cell bodies of submucous neurons varied from 0-4 and the number of VIP-immunoreactive close contacts varied from 3-10. There was no significant difference between VIP-immunoreactive nerve cell bodies and non-VIP nerve cell bodies in the number of VIP-immunoreactive synapses and close contacts they received. Thus, the implication from light microscopy that NOS/VIP terminals end predominantly on VIP nerve cells was not vindicated by electron microscopy.

Histochemical and immunohistological methods have been used to demonstrate the presence of nitric oxide synthase (NOS) in the intracardiac parasympathetic ganglion of bullfrog heart. Both NADPH-diaphorase staining and indirect immunofluorescence with polyclonal antibodies raised against the C-terminal regions of a cloned NOS from rat cerebellum show that the postganglionic, parasympathetic neurons in the intracardiac ganglion contain NOS. This suggests that nitric oxide may have a role in neuronal activity in frog heart.

Nitric oxide synthase (NOS)- and vasoactive intestinal peptide (VIP)-immunoreactive neurons projecting to the upper rectum or penis were examined using retrograde tracing combined with immunohistochemistry in the major pelvic ganglion of male rats. Five days after injection of Fluoro-Gold (FG) into the upper rectum or penis, the major pelvic ganglion was treated with colchicine. FG injected into the upper rectum labelled many ganglion neurons in the major pelvic ganglion. Immunohistochemistry showed that 37% of FG-labelled neurons were immunoreactive for NOS and 33% for VIP. After injection of FG into the penis, 41% of FG-labelled neurons were immunoreactive for NOS and 25% for VIP. Serial cryostat sections stained for NOS and VIP, respectively, showed the co-localization of NOS and VIP in the ganglion cells projecting to the rectum and penis. In the major pelvic ganglion of the colchicine-treated animals, about 17% of the ganglion cells were immunoreactive for NOS and 32% were immunoreactive for VIP. These neurons were small in diameter (less than 30 microns). A histogram showing cell sizes in cross-sectional areas of NOS-immunoreactive neurons coincided with that of VIP-immunoreactive neurons. Most of the NOS- and VIP-immunoreactive neurons were less than 600 microns. These results indicate that small neurons containing both NOS and VIP in the major pelvic ganglion project to the rectum and penis. In the penile erectile tissues and enteric ganglia, NO and VIP may be released from the same axons and may act concomitantly on the target tissue.

Intracellular recordings were made from 73 guinea-pig submucosal neurons using neurobiotin-filled microelectrodes; subsequently, neuropeptide immunoreactivity, morphology and nerve fibre projections were determined. Five distinct groups of cells could be distinguished: S cells with inhibitory input (61%), S cells without inhibitory input (19%), AH cells (8%), S-AH cells (5%), and glial networks. S cells with inhibitory input were immunoreactive for vasoactive intestinal polypeptide and showed Dogiel Type III morphology with the axon branching and coursing through two to 12 ganglia; varicosities and tufts of varicosities were observed surrounding other cell bodies as well as over blood vessels. S cells without inhibitory input primarily were immunoreactive for neuropeptide Y; they also showed Dogiel Type III morphology and similar, though shorter, axonal projections and varicose features surrounding other neurons. AH cells, which most likely contained substance P, lacked synaptic input and exhibited Dogiel Type II morphology; they branched more extensively than S cells and also formed varicose tufts within other ganglia. S-AH cells combined electrophysiological properties of S cells with inhibitory input and AH cells and did not show consistent morphological or histochemical characteristics. Typical glial networks were observed; in addition, on two occasions unusual networks of dye and electrical coupling between S cells without inhibitory input and a glial complex were observed. These results suggest that vasoactive intestinal polypeptide-containing S cells may act as interneurons which mediate a slow excitatory synaptic potential; that neuropeptide Y-containing S cells, which are known to be cholinergic, may play a role as cholinergic interneurons mediating the nicotinic fast excitatory synaptic potential; and that AH neurons also may provide cholinergic innervation to other submucosal neurons in addition to their previously described dual projections into mucosa and myenteric plexus.

The distribution, pathways and origins of peptide-containing nerve fibers in the anterior two thirds of the dog tongue were investigated using immunohistochemistry combined with retrograde axonal tracing and denervation experiments. Within the epithelium of the fungiform papillae, varicose nerve fibers immunoreactive to substance P (SP) and calcitonin gene-related peptide (CGRP) were present. These disappeared completely after severance of the lingual nerve (LN) alone. Dense CGRP-immunoreactive varicose fibers surrounded cell bodies in the intralingual ganglia (ILG), which consisted of neurons immunoreactive to vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY) and SP. These CGRP-immunoreactive fibers disappeared following severance of the chorda tympani (CT) alone. SP-, CGRP-, VIP-, NPY- and tyrosine hydroxylase (TH)-immunoreactive nerve fibers were distributed around the walls of blood vessels, especially arteriovenous anastomoses (AVAs). None of these immunoreactive fibers completely disappeared after severance of the LN or CT alone, but SP- and CGRP-immunoreactive fibers disappeared following severance of both the LN and CT. TH-immunoreactive fibers disappeared after ganglionectomy of the superior cervical ganglion (SCG) or severance of the hypoglossal nerve (HGN). VIP- and NPY-immunoreactive fibers invariably remained after various denervation experiments. In tracing experiments, CGRP-immunoreactive as well as SP and CGRP-immunoreactive cells in the trigeminal ganglion were labelled from the LN, and those in the geniculate ganglion and jugular ganglion were labelled from the CT. A large number of neurons in the SCG were labelled from the HGN, with some of these being SP and CGRP-immunoreactive. These results demonstrate that SP- and CGRP-immunoreactive fibers from the trigeminal ganglion are distributed to the lingual epithelium; vascular walls receive SP- and CGRP-immunoreactive sensory fibers from the LN as well as CT, some SP and CGRP-immunoreactive fibers from the SCG in addition to catecholaminergic sympathetic fibers, and VIP- and NPY-immunoreactive parasympathetic fibers from the ILG. The ILG is also considered to be innervated by CGRP-immunoreactive sensory fibers from the CT.

Neurofilaments (NF) are composed of three polypeptides of differing molecular size, termed NF-L, NF-M and NF-H. The NF-H and, to a lesser degree, NF-M components are phosphorylated. In the majority of normal neurones, the location of phosphorylated NF is confined to neuronal processes, particularly the axon, and excluded from the perikaryon. Cell bodies of autonomic neurones of the rat do not contain phosphorylated NF. In many disease states, phosphorylated NF accumulate in the neuronal cell body and therefore in most circumstances their presence indicates abnormality. This paper reports that in at least two autonomic ganglia of normal horses (stellate and coeliomesenteric) the vast majority of neuronal perikarya immunostain strongly for phosphorylated NF. Pretreatment with alkaline phosphatase abolishes staining.

Albino mice, infected with Trypanosoma cruzi (Tulahuen strain) were sacrificed on days 7, 9, 12, 14, 16, 18, 21, 32 and 39 following infection. Transmission electron microscopic examination of the cardiac ganglia revealed no ultrastructural change at day 7. At day 9 there was peri- and intraganglionic monocytic infiltration but parasites were absent. Between days 12 and 16 there was intense monocytic infiltration, with intra-ganglionic presence of parasites within fibroblasts, monocytes and macrophages. None were seen within capsular cells, endothelial cells, Schwann cells, satellite cells and ganglion cells. The Schwann cells and satellite cells, however, showed phagocytic activity. Satellite cells were also reactive with proliferative pseudopodia which encircled neuronal processes. By day 18, parasites were absent in the ganglia. But monocytes were still present up to day 39, some of them still engulfing satellite cell and neuronal processes. Satellite cells continued to be reactive and Schwann cells phagocytic. Ganglion cells remained normal throughout the experiment. The results suggest that infection of Schwann cells, satellite cells and ganglion cells may depend upon the tissue tropism of the strain of the parasite used and its concentration in the inoculum. The results are consistent with the view that any parasympathetic dysfunction in experimental Chaga's disease in the mouse may be of a transient nature.

Data, received in investigation of the small intestine intramural ganglia of 34 suckling rabbits show, that development of experimental cholera induce in the neurons reactive shifts, changed by dystrophic alterations. Prevaluation of cholinergic influences in autonomic ganglia in small intestine promotes enterocytes hypersecretion and accelerating of motor activity.

Ultrastructure of the neurons of the pterygopalatine ganglion in two groups of rats with body temperature reduced to 22 degrees C and 18 degrees C of rectal temperature for periods of 3 hours, was examined. In both experimental groups various degree of cytoplasmic dispersion and widening of endoplasmic reticulum and cisterns of Golgi complex were observed. Nuclear envelope was often folded. In some mitochondria reduction of their crests and density of matrix and swelling were visible. Enlargement of the endoplasmic reticulum in the glial cells were also observed. The degree of ultrastructural changes in the group of animals cooled to the rectal temperature of 18 degrees C were more pronounced than in the group cooled to 22 degrees C.

A series of five patients with a variant type of Jansky-Bielschowsky disease is presented. The disease initiated between four and a half and seven years with mental and slight motor symptoms. The additional neurological symptoms and signs, i.e. visual failure, retinal degeneration, ataxia, myclonia and epilepsy developed in all children before the age of ten years. The present series differs from our previous series of 16 cases especially in regard of neurophysiological findings (photic spikes, high visual evoked potential, VEP and high somatosensory evoked potential, SEP). VEP became abnormally high between 8.0 and 9.5 years instead of being an early finding as in the previous series. Photic spikes appeared also later in the present series. Electromicroscopic investigation revealed cytosomes with fingerprint profiles (FP) in the autonomic ganglion cells and cytosomes with both FP and curvilinear (CP) profiles in many extraneural cells including smooth muscle, Schwann cells, capillary endothelium and macrophages. In the light of our 21 Finnish patients and the literature, the spectrum of Jansky-Bielschowsky disease seems to be much wider than previously assumed. The diagnosis should be based on clinical, ophthalmological, neurophysiological and ultrastructural findings. Repeated neurophysiological studies may be necessary.

The external limiting membrane of the avian embryonic optic tectum is isolated by mechanically separating the neuronal mesencephalon from the overlying mesenchymal tissue. The preparation consists of a basal lamina which is covered on its neural side by endfeet of neuroepithelial cells and has attached to it on its meningeal side a collageneous stroma, containing blood vessels. The external limiting membrane can be flat-mounted on a piece of nitrocellulose filter as mechanical support. It covers an area between 0.3 and 1 cm2, depending on the age of the donor embryo. The endfeet can be removed together with all cellular components of the meninges by treatment with 2% Triton-X-100 or with distilled water. The basal lamina itself is approximately 80 nm thick and consists of two laminae rarae and a central lamina densa. Immunohistochemical staining reveals that the basal lamina in the embryo, after isolation and after detergent extraction of the isolated preparation, contains type IV collagen, nidogen, laminin, and low density heparan sulfate proteoglycan as do other basement membranes. Antibodies against the neural cell adhesion molecule (N-CAM), chondroitin sulfate proteoglycan, and fibronectin fail to stain the external limiting membrane, but these proteins were clearly identified in the blood vessel-containing meninges or in the optic tectum. The flat-mounted external limiting membrane preparation was used as substrate to culture several different neural tissues of central and peripheral origin. Explants of neural crest cells, dorsal root ganglia, and sympathetic ganglia can be cultured on the external limiting membrane. All explants grow well on the basal lamina preparations whether the endfeet are attached or detergent-extracted prior to explantation; however, neurite outgrowth from sympathetic ganglia is reduced in the presence of the endfeet. Although the endfoot-lined external limiting membrane represents at least part of the immediate environment encountered by retinal axons as they invade the optic tectum and despite its excellent properties as a substrate for retinal axons in vitro, cues guiding the orientation of axons were not detected in the flat-mounted preparation.