In vitro effects of histamine liberator compound 48/80 on rat superior cervical ganglia with special regard to the small granule-containing cells

Summary The effect of the histamine liberator compound 48/80 on the rat superior cervical ganglia in vitro has been investigated. After incubation of the ganglia with compound 48/80: (1) ganglionic mast cells degranulate in the same manner as in other tissues; (2) cell bodies of the postganglionic neurons are not affected by compound 48/80; (3) there is evidence that ganglionic interneurons, the monoamine-containing cells are activated. The ultrastructural aspects of this process are characterized by degranulation of perikarya and accumulation of dense core vesicles in cell processes and in terminals adjacent to presynaptic membranes. These vesicles vary in size between 200–800 Å in diameter. They may represent storage sites of the neurotransmitter complexes that have undergone exocytosis. The results are discussed with special reference to models of exocytotic processes involving the adrenergic transmitter. It is concluded that monoamine-containing cells represent interneurons in sympathetic ganglia which inhibit ganglionic transmission and which are stimulated by low concentrations of compound 48/80 in vitro.This work was supported by the Deutsche Forschungsgemeinschaft (SFB 70 Hirnforschung).The authors wish to thank Professor Dr. J. Staubesand for his encouragement in the course of this work.Dedicated to Prof. Gian Töndury, Zurich, on the occasion of his 70th birthday.     

Stimulation of brain mast cells by compound 48/80, a histamine liberator, evokes renin and vasopressin release in dogs

Because degranulation of brain mast cells activates adrenocortical secretion (41, 42), we examined whether activation of such cells increases renin and vasopressin (antidiuretic hormone: ADH) secretion. For this, we administered compound 48/80 (C48/80), which liberates histamine from mast cells, to pentobarbital-anesthetized dogs. An infusion of 37.5 microg/kg C48/80 into the cerebral third ventricle evoked increases in plasma renin activity (PRA), and in plasma epinephrine (Epi) and ADH concentrations. Ketotifen (mast cell-stabilizing drug; given orally for 1 wk before the experiment) significantly reduced the C48/80-induced increases in PRA, Epi, and ADH. Resection of the bilateral splanchnic nerves (SPX) below the diaphragm completely prevented the C48/80-induced increases in PRA and Epi, but potentiated the C48/80-induced increase in ADH and elevated the plasma Epi level before and after C48/80 challenge. No significant changes in mean arterial blood pressure, heart rate, concentrations of plasma electrolytes (Na+, K+, and Cl-), or plasma osmolality were observed after C48/80 challenge in dogs with or without SPX. Pyrilamine maleate (H1 histaminergic-receptor antagonist) significantly reduced the C48/80-induced increase in PRA when given intracerebroventricularly, but not when given intravenously. In contrast, metiamide (H2 histaminergic-receptor antagonist) given intracerebroventricularly significantly potentiated the C48/80-induced PRA increase. A small dose of histamine (5 microg/kg) administered intracerebroventricularly increased PRA twofold and ADH fourfold (vs. their basal level). These results suggest that in dogs, endogenous histamine liberated from brain mast cells may increase renin and Epi secretion (via the sympathetic outflow) and ADH secretion (via the central nervous system).     

The effects of 17β-oestradiol, testosterone and tamoxifen on the development of papillomata in Catostomus commersoni

Testosterone induced papillomata in 85% (11/13) of initially non-papillomatous white suckers Catostomus commersoni and increased papillomata growth in 100% (16/16) of initially papillomatous suckers. 17β-oestradiol induced papillomata in 83% (10/12) of initially nonpapillomatous suckers and increased papillomata growth in 100% (16/16) of initially papillomatous suckers. Less than 29% (4/14) of white suckers injected with tamoxifen developed papillomata, while complete papillomata regression was observed in 71% (10/14) of initially papillomatous suckers. In control groups 59% (27/46) of suckers either retained or developed papillomata and 27% (6/24) of suckers exhibited tumour regression. Protein kinase C (PKC) activity was significantly depressed and ornithine decarboxylase (ODC) activity was significantly elevated in steroid-treated papillomata v. normal lip epidermis. ODC activity was significantly depressed in tamoxifen-injected, regressing papillomata. There were no significant differences in plasma oestrogen and testosterone levels between papillomatous and nonpapillomatous female fish from a site contaminated with persistent organic chemicals and an uncontaminated reference site. Similarly, no significant differences in testosterone and 11-ketotestosterone levels were observed between papillomatous and non-papillomatous male fish.     

Effects of compound 48/80 and exogenous histamine on wound healing in mice

Compound 48/80 has previously been shown to improve wound healing in rats, presumably through stimulation of histidine decarboxylase activity and mobilization of histamine from mast cells. In the present study, C57Bl/6 mice were wounded by dorsal skin incision followed by treatment with compound 48/80, exogenous histamine, or the combination of 48/80 plus histamine. Skin-breaking strength was significantly increased over saline-injected controls by the combined treatment with 48/80 and histamine. Neither 48/80 or histamine alone had any influence on wound healing. Histamine content of skin at the wound site was significantly reduced by 48/80 treatment, but was unaffected by 48/80 plus histamine or histamine given alone. In contrast, stomach and leg muscle histamine levels were significantly increased beyond those of unwounded, wounded saline- or 48/80-injected mice. These results were also confirmed in CD mice, and are in contrast to findings in rats in which treatment with 48/80 alone significantly improved wound healing of similarly injured animals.     

Compound 48/80 induces endothelium-dependent and histamine release-independent relaxation in rabbit aorta.

Compound 48/80 (C48/80) is a synthetic condensation product of N-methyl-p-methoxyphenethylamine with formaldehyde and is an experimental drug used since the 1950s to induce anaphylactic shock through histamine release. This study was carried out to further elucidate the mechanism by which this drug induces nitric oxide (NO) release. Our specific goals were: (a) to verify if C48/80's relaxation occurs through the stimulation of histamine receptors; (b) to evaluate the endothelium-dependent relaxation induced by C48/80; (c) to identify NO as the endothelium-relaxing factor released by C48/80; (d) to identify the NO synthase (NOS) responsible for NO release; and (e) to verify if the relaxation induced by C48/80 is calcium and cyclic guanidine monophosphate (cGMP) dependent. Rabbit aorta segments, with and without endothelium, were suspended in organ chambers (25ml) filled with Krebs solution maintained at 37 degrees C, bubbled with 95% O(2)/5% CO(2) (pH 7.4). Phenylephrine was used to contract the segments. Other protocol drugs included H(1)- and H(2)-receptor antagonists, cyclooxygenase, NOS, guanylyl cyclase and phospholipase C (PLC) inhibitors. Endothelium-dependent relaxation induced by C48/80 was also studied in calcium-free Krebs solution associated with a calcium chelator. In summary, our investigation demonstrated that the C48/80 vasodilating action: (a) does not depend on H(1) and H(2) histamine receptors; (b) is NO endothelium-dependent; (c) is dependent on the endothelial constitutive NOS (NOS-3) isoform activation; (d) is cGMP-dependent; and that NOS-3 activation by C48/80: (a) is independent of PLC up to 25mug/ml and (b) is partially dependent of this lipase in higher doses.     

Regression,proliferation and development of lip papillomas in wild white suckers,Catostomus commersoni,held in the laboratory

Synopsis Epizootics of skin neoplasms in teleost species have been documented within the Great Lakes region over the last decade. The white sucker,Catostomus commersoni, has been proposed as a sentinel species to monitor environmental health in these systems. The prevalence of skin neoplasia is elevated in white suckers and other fish taken from chemically polluted sites or from lake regions adjacent to heavy industry. Lip papillomas regress and proliferate spontaneously in captive wild suckers. It is important to investigate the relevance of these observations to papilloma etiology to determine whether the prevalence of this disease is a suitable biomarker for environmental health. White suckers were captured in the spring of 1992 and 1993 during annual spawning runs (mid to late April) at the Ganaraska River, which discharges into Lake Ontario at Port Hope, Ontario. Under crowded laboratory conditions, there was either proliferation of existing papillomas or development of new papillomas. However, in uncrowded conditions, existing papillomas either regressed completely or there was no development of new papillomas. Protein Kinase C (PKC), a proposed marker enzyme for hyperplasia and neoplasia, was used to determine if regressing and proliferating papillomas could be differentiated on the basis of biochemical activity. PKC activity was lower in proliferating papillomas, but not significantly different from papillomas sampled initially from Ganaraska River suckers. Regressing papillomatous tissue displayed a significantly higher level of enzyme activity than either proliferating or unchanged papillomas, but the PKC activity of regressing papillomas was not significantly different from that of normal lip epidermis.     

Induction of histamine release and calmodulin antagonism are two distinct properties of compound 48/80

Compound 48/80, a mixture of oligomers, was fractionated by passing it in the presence of Ca2+ over a calmodulin-Sepharose column. The fraction not retained by the gel was shown by mass spectrometry to consist mainly of trimers, tetramers and pentamers. A second fraction consisting of hexamers and heptamers was eluted from the column at high ionic strength in the presence of Ca2+. Finally, in the presence of EGTA at high ionic strength, a third fraction eluted mainly consisting of higher oligomers (hexamers to dodecamers). The different fractions were characterized by testing their influence on calmodulin-sensitive Ca2+-transporting ATPase and their ability to elicit histamine release from mast cells. The third fraction showed the highest potency as calmodulin antagonist, however, the second fraction was the most potent in inducing histamine secretion. This would imply that the ability of compound 48/80 to evoke histamine release and to inhibit the function of calmodulin are distinct properties of the agent which are unrelated.