[Biogenic amines in the ganglia of the autonomic nervous system in emotional stress]. / Biogennye aminy v gangliiakh avtonomnoi nervnoi sistemy pri émotsional'nom stresse.

Rabbits with different stability of cardiovascular functions were shown to have different activity of sympathetic nervous system under emotional stress. A small enhancement of the system activity occurred in animals with practically unchanged arterial pressure, whereas its hyperactivity was found in the animals with a progressive decrease of arterial pressure. Different changes of catecholamines in the nodose ganglia of these two groups of animals seem to reflect the participation of those ganglia in the maintenance of stable cardiovascular functions in extreme conditions.

[Immunohistochemical studies of putative neurotransmitters in the lower urinary tract].

Putative neurotransmitters of the lower urinary tract were investigated in apes, rabbits and cats using immunohistochemical techniques of PAP (Peroxidase antiperoxidase) staining and IGSS (Immunogold silver staining) methods for Neuron specific enolase (NSE), Acetylcholine (Ach), Noradrenaline (NA), Vasoactive intestinal polypeptide (VIP), Substance P (SP) and Calcitonin gene related peptide (CGRP). 1) The localization of pelvic ganglions exhibited more striking evidence of species difference. Huge pelvic ganglions were found particularly in the dorsolateral area of the prostate in apes. On the other hand, in cats and rabbits, many ganglion cells were found around the uretero-vesical junctions. 2) In the pelvic ganglions of the apes, Ach immunoreactives were detected in nearly 70 percent of the cell bodies. 10-15 percent were NA immunoreactive cells. In addition, 15-20 percent VIP and a smaller percentage of SP immunoreactive cells were detected in the same ganglions. Axons extending from the ganglion cells showed the intense neurotransmitters immunoreactivity. 3) In the apes, varicose fibers containing SP were widely distributed in the epithelium, submucosa, muscle layer, and around the vessels of the bladder. SP immunoreactive cell bodies were found in the dorsal root ganglion at levels of L7, S1 and at the same levels in the posterior horn. On the other hand, the bulbourethral gland and the seminal vesicle contained SP immunoreactive cell bodies. 4) CGRP containing fibers were distributed in similar locations as SP containing fibers in the bladder. 5) VIP immunoreactive fibers were also widely distributed, being most dense at the base of the bladder.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunohistochemical localization of parvalbumin in rat and monkey autonomic ganglia.

The cellular distribution of parvalbumin-like immunoreactivity in autonomic ganglia such as superior cervical sympathetic ganglia, paravertebral sympathetic chain ganglia (T6), ciliary ganglia and enteric ganglia was investigated by immunohistochemical peroxidase-antiperoxidase methods using an antiserum against rat skeletal muscle parvalbumin. We detected parvalbumin-like immunoreactivity in almost all neurons of rat superior cervical sympathetic ganglia and other paravertebral sympathetic chain ganglia, where the antigen was located in the cytoplasm but the nuclei were not labelled. No neurons positive for parvalbumin-like immunoreactivity were observed in rat ciliary ganglia or enteric ganglia. In monkey, almost all neurons of the superior cervical sympathetic ganglia contained parvalbumin-like immunoreactivity, but none of the neurons of the ciliary ganglia were labelled with the antiserum to parvalbumin. These results suggest that parvalbumin-like immunoreactivity exists in a specific subpopulation of the neurons of the autonomic nervous system.

Substance P-like immunoreactive innervation of the human colon and coeliac, superior mesenteric and inferior mesenteric ganglia.

The localization of substance P is studied in the human prevertebral sympathetic ganglia and descending colon by the indirect immunofluorescence technique. Strongly immunoreactive axons reach the coeliac, superior mesenteric and inferior mesenteric ganglia. The sympathetic postganglionic neurones are surrounded by a meshwork of positive beaded fibers and terminals. Varicose nerve fibers are variously distributed in all nonepithelial layers of the colon. A close network of fibers and terminal surrounds the ganglionic cells in the mienteric and submucous plexuses. Substance P-like immunoreactive perikarya are present in these structures.

Distribution of substance P immunoreactive cell bodies and fibers in cranial sensory and autonomic ganglia of the chick.

Substance P-immunoreactive neurons were demonstrated in chick embryonic and adult trigeminal ganglion and jugular-superior ganglionic complex using FITC-immunohistochemical methods. Both small-size and large ganglion cells exhibited SP immunoreactivity, without apparent changes during embryonic and post-hatching development. SP-positive fibers could be detected in a good number in the sympathetic cranial cervical ganglion, either during embryonic development or in adult chick. No immunoreactive perikarya were observed in this ganglion. In the ciliary ganglion, both choroidal and ciliary neurons were SP-negative, whereas SP immunoreactive fibers surrounded the perikarya of both cell populations.

Autonomic neurons supplying the rat eye and the intraorbital distribution of vasoactive intestinal polypeptide (VIP)-like immunoreactivity.

We traced the origin and path of autonomic nerves to the rat eye using, as an aid to dissection, a modified thiocholine method for the histochemical demonstration of cholinesterase. When applied to whole nerves and ganglia supplying the rat eye, this procedure is not specific for cholinergic neurons; instead it stains both sympathetic and parasympathetic nerves, many of which are otherwise too fine to identify in dissection. We found that nerves from the superior cervical and pterygopalatine ganglia form a plexus at the orbital apex corresponding to the retro-orbital plexus found in rabbit, monkey and man. In the rat, nerves from the retro-orbital plexus travel peripherally to the superior surface of the optic-nerve sheath. Here, they fuse with long ciliary nerves and the post-ganglionic nerves from the ciliary ganglion to form another dense nerve-fiber plexus that ultimately supplies the eye. Importantly, the plexus on the optic nerve contains many isolated or aggregated ganglion cells. These are comparable in number to those in the ciliary ganglion itself and are assumed to be accessory ciliary neurons. Using immunohistochemistry, we also sought evidence for vasoactive intestinal polypeptide in these ganglia and nerves. As previously known, many pterygopalatine ganglion cells are immunoreactive for this peptide. Vasoactive intestinal polypeptide (VIP)-like immunoreactive nerve fibers are present in nerves from the retro-orbital plexus to the optic-nerve sheath plexus, in most nerves of the latter plexus, and in most nerves entering the eye. Furthermore, a small proportion of nerve cells in the main and accessory ciliary ganglia also are immunoreactive for VIP. We conclude that in addition to the pterygopalatine ganglion, the ciliary ganglion and its accessory ganglia are sources of VIP-like immunoreactive nerves in the rat eye.

S-100 protein expression in satellite and Schwann cells in neuroblastoma. An immunohistochemical and ultrastructural study.

Immunohistochemical evidence has recently been provided that in the normal adrenal medulla as well as in autonomic ganglia, satellite cells and Schwann cells react with S-100 protein antiserum. In the light of these data, we investigated primary peripheral neuroblastoma and ganglioneuroblastoma to determine firstly whether both cell populations actually exist in the malignancies, using the definite criteria of electron microscopy for their identification, and secondly whether they express S-100 protein using on immunohistochemical technique and light microscopy. The results indicate that in both neuroblastoma variants, satellite and Schwann cells are present and specifically express the S-100 antigen.

ANF-like peptide(s) in the peripheral autonomic nervous system.

The recent demonstration of the atrial natriuretic factor (ANF) within the brain has been extended in the present study by the additional localization of ANF-like activity in the peripheral nervous structures. Using a sensitive radioimmunoassay, it was possible to detect ANF-like immunoreactive peptide(s) in crude and chromatographically separated extracts of parasympathetic rat ganglia. The partially purified ANF-like peptide exhibited a biological action similar to cardiac ANF. This finding supports a possible involvement of ANF in the regulation of both, central and peripheral neuronal activities.

Distribution of calcitonin gene-related peptide in the rat peripheral nervous system with reference to its coexistence with substance P.

This immunocytochemical study, using a double-staining method, showed that calcitonin gene-related peptide-like immunoreactive structures are widely distributed in the peripheral nervous system and that many of them coexist with substance P-like immunoreactive structures in single sensory ganglion cells. Neurons positive for calcitonin gene-related peptide but negative for substance P were detected in sensory ganglia. These cells were large (about 30-45 micron in diameter); these primary sensory neurons containing calcitonin gene-related peptide can probably act independently of substance P. There were neurons containing calcitonin gene-related peptide without substance P in the pterygopalatine ganglion, although these cells were less numerous than in the sensory ganglia. In consecutive sections, calcitonin gene-related peptide-like structures occurred in thyroid parafollicular cells, which also contain calcitonin. This suggested that messenger RNA for producing calcitonin gene-related peptide is also present in the thyroid, and like calcitonin, calcitonin gene-related peptide may have a peripheral physiological role.

Pancreatic ganglioneuronal amyloid. Occurrence in diabetic cats with islet amyloidosis.

Amyloid in pancreatic ganglia and nerves (ganglioneuronal amyloid) was demonstrated in 4 of 8 diabetic cats with islet amyloid deposits. Eighteen nondiabetic cats (including 4 with islet amyloid) did not have detectable amyloid in pancreatic nerves or ganglia. Ganglioneuronal amyloid had staining characteristics identical to those previously reported for islet amyloid, including 1) congophilia, 2) resistance to oxidation by KMnO4, 3) immunoreactivity (PAP technique) with antiserum to a B-chain-rich insulin fraction, and 4) no reactivity with antisera to insulin, glucagon, or somatostatin. Nonneuronal cells with insulin, glucagon, and somatostatin immunoreactivity were seen in many pancreatic ganglia and nerves; and in a few instances, B cells were found near ganglioneuronal amyloid deposits. The premise that these ganglioneuronal amyloid deposits (like islet amyloid) are insulin-related is supported by their immunoreactivity with antiserum to B-chain-rich insulin and the demonstration of B cells in pancreatic ganglia and nerves.

Intramural ganglia of the human urinary bladder.

The arrangement and distribution of intramural autonomic ganglion cells have been examined in samples of the dome and lateral walls of the human urinary bladder. Ganglia were frequently observed in samples removed from either site and possessed histochemical characteristics to support their classification as presumptive cholinergic neurons. Unlike pelvic autonomic neurons, intramural bladder ganglion cells are not associated with noradrenergic (possibly inhibitory) preganglionic nerve terminals. The widespread distribution of ganglion cells within the bladder wall serves to frustrate surgical attempts to denervate detrusor smooth muscle. The outcome of such operative procedures is likely to result in decentralisation rather than denervation of the urinary bladder.

A quantitative study of acetylcholine in Hirschsprung's disease.

This is the first report on acetylcholine (Ach) quanititaion of the aganglionic colon by a chemical assay. Ach content in the colon of Hirschsprung's disease was measured by pyrolysis gas chromatography, which was found to be a simple, accurate, and reliable method. Microwave irradiation on the surgical resected specimens was performed before extraction procedure to abolish acetylcholinesterase (Ach-E) and cholineacetyltransferase (CAT) activity in the tissue. Ach content in the ganglionic colon was 8.51 +/- 3.15 nmole/g (n 5 in contrast to 23.79 +/- 14.17 nmole/g (n = 19) in the aganglionic segment of Hirschsprung's disease. The meaning of elevated Ach level in the aganglionic segment was discussed from the pathophysiologic standpoint.

Vasoactive intestinal peptide and its relationship to ganglion cell differentiation in neuroblastic tumors.

Immunohistochemical studies have demonstrated that immunoreactive vasoactive intestinal peptide is present in, and restricted to, the differentiating and mature ganglion cells in a variety of normal and neoplastic neural tissues. In a composite pheochromocytoma-ganglioneuroma (associated with the syndrome of watery diarrhea, hypokalemia, and hypochlorhydria), five ganglioneuroblastomas, five ganglioneuromas (two of which were associated with diarrheal syndromes), an unusual mixed neuroblastoma-ganglioneuroma, and four normal sympathetic ganglia, vasoactive intestinal peptide was present in differentiating and mature ganglion cells. The peptide was also demonstrated in isolated ganglion cells in two pheochromocytomas but was not present in pheochromocytes, Schwann cells, or undifferentiated neuroblastic cells in the neuroblastomas and ganglioneuroblastomas. These studies indicate that the presence and presumably the production of vasoactive intestinal peptide thus reflect a particular line of neuroblastic differentiation and are not merely a reflection of common derivation of these tissues. Our identification of vasoactive intestinal peptide in neurogenic tumors associated with diarrhea supports the contention that the peptide might be an important diarrheogenic factor in these tumors.

Vacuolated neurons in the hypogastric ganglion of the rat.

The vacuolated neurons (VN) of the main hypogastric ganglion of the male rat were studied using the formaldehyde-induced fluorescence (FIF) method for the histochemical demonstration of catecholamines. Microspectrofluorimetry was performed to identify the fluorophores and to quantify the FIF. The thiocholine method (Koelle-Gomori) was used to demonstrate acetylcholinesterase activity. The fine structure of the VN was studied using glutaraldehyde/OsO4 fixation. (1) In the untreated adult male rat VN represent only a small population of the total number of hypogastric neurons (0.8--1.2%). The vacuoles are similar to those of the VN from the corresponding female ganglion. (2) The VN are considered to be adrenergic due to the nature of their fluorophore, indicating a primary catecholamine. (3) The first VN appear in the hypogastric ganglia at the age of 7 weeks. After testosterone administration to young rats, VN are found at the age of 4 weeks. (4) The basic fine structure of the VN is similar to that of other ordinary neurons of the hypogastric ganglia. (5) The content of the vacuoles could not be identified. (6) Indications of degeneration were not observed in the VN. (7) The VN are interpreted as being a functional stage of the "short" adrenergic neurons, which are under the control of steroid hormones. (8) Fifteen months after castration, no VN could be found in the hypogastric ganglia, while their number was normal in the corresponding control animals.

[Effect of histological processing on the nucleic acid, free nucleotide and protein content in the cranial cervical sympathetic ganglion]. / Vliianie gistologicheskoi obrabotki na soderzhanie nukleinvykh kislot, svobodnykh nukleotidov i belka v kranial'nom sheinom simpaticheskom ganglii.

By means of biochemical techniques, the stability of free nucleotide contents has been demonstrated in the rabbit sympathetic node fixed in the Carnoy solution for 1--2 hours at 4 degrees C. A 2 hour fixation at room temperature and at 37 degrees, however, results in a considerable loss of free nucleotides--of the total amount in the fresh ganglion tissue, resp., 66.6 and 74.5%. According to these data, the Carnoy solution can be successfully used for a quantitative precipitation of free nucleotides. A histological treatment of this Carnoy fixed sympathetic ganglion (for 2 hours, at 4 degrees) causes no losses of nucleic acids, free nucleotides or protein, does not affect RNA, DNA or protein contents, but leads, however, to a remarkable loss of free nucleotides--up to 45%.

[Bioassay for acetylcholine from the rat cervical sympathetic ganglion (author's transl)]. / Une méthode de dosage de l'acétylcholine appliquée à l'étude du ganglion sympathique cervical du rat.

1. A bioassay for acetylcholine (Ach), with a sensitivity of 0.1 picomoles in 30 microliters (microliter) (Fig. 3), was developed using the powerful musculature of the common leech (Haemopis sanguisuga). 2. Ach was extracted from a pool of ganglia with sodium tetraphenylborate in 3-heptanone and analysed using gas chromatography. The chromatograph confirmed that the "Ach like effect" measured with bioassay was due to acetylcholine and not to one of its derivatives (Fig. 5 and 6). 3. An incubation chamber for perfusing the isolated superior cervical ganglion of the rat is described, with which it is possible to do electrophysiological experiments and to sample the Ach lost or released by the ganglion (Fig. 2). 4. Using this system, the results obtained from the resting or stimulated rat ganglion incubation in-vitro, were similar to those obtained with the cat ganglion perifused in situ. 5. Experiments were also performed to investigate the effect of removing the two precursors of Ach, glucose (Table I) and choline (Fig. 9), from the incubation medium.