The need of protein synthesis for the effect of LH--RH on the cAMP level in the anterior pituitary in vitro

The effect of LH--RH on cAMP accumulation in the anterior pituitary of male rats was investigated. The effect consisted of two phases. In the first phase of incubation (about 2 hours), there was no change in the cAMP level, while in the second phase a significant increase was observed. Cycloheximide (10(-5)M) given simultaneously with the LH--RH totally blocked the observed effect of LH--RH. If cycloheximide was given in the second phase of incubation, when the cAMP level was already elevated, its further rise was prevented and the elevated cAMP level remained unchanged for 2 or more hours. Actinomycin D (10(-5)M) added together with LH--RH totally abolished the action of the latter, but if actinomycin D was given 1 hour after LH--RH a significant increase of the cAMP level was found at the end of the 4-hour incubation.     

The increase of cerebellar cAMP level after decapitation: the effect of propranolol

The increase of cAMP level of the rat cerebellum induced by decapitation was studied. Administration of 5 mg/100 g propranolol 1 hour before decapitation completely prevented this increase. Neither the depletion of catecholamine pools, inhibition of their synthesis, nor barbiturate treatment influenced the increase of cAMP level evoked by decapitation. It has been concluded that noradrenergic neurotransmission is involved in the cerebellar cAMP level increase after decapitation.     

Lipopolysaccharide receptor on rabbit peritoneal macrophages. I. Binding characteristics

The Bordetella pertussis endotoxin, labeled with tritium ((3H)-LPS), bound irreversibly and nonspecifically to rabbit lung macrophages, but bound reversibly and specifically to both resident and elicited rabbit peritoneal macrophages. The specific binding capacity of the macrophages was saturated with about 3 X 10(4) LPS molecules per cell. The binding was inhibited with the homologous unlabeled endotoxin, but not at all with endotoxin from Proteus mirabilis, thus assessing ligand specificity. Endotoxins from other bacteria gave intermediate inhibition value. Binding of tritium-labeled pertussis endotoxin was significantly inhibited by one of the two polysaccharides (PS-1) present in this endotoxin, but neither the other polysaccharide (PS-2) nor the Lipid A fragment exhibited such activity. These results strongly suggest the presence of a lectin-like receptor for LPS on the membrane of rabbit peritoneal macrophages.     

A novel type of endotoxin structure present in Bordetella pertussis. Isolation of two different polysaccharides bound to lipid A.

The endotoxin of Bordetella pertussis was cleaved by mild acidic hydrolysis to yield a polysaccharide (polysaccharide I, 15%), a glycolipid (63%) and lipid X (2%). Further treatment of the glycolipid with stronger acid released a second polysaccharide (polysaccharide II, 9%) and material similar to lipid A present in enterobacterial endotoxins. Both polysaccharides possess a single molecule of 3-deoxy-2-octulosonic acid as the reducing, terminal sugar. In polysaccharide II the octulosonic acid is phosphorylated in position 5 and presumably substituted in position 4; in polysaccharide I the octulosonic acid is not phosphorylated, but is substituted in position 5. Following treatment of the endotoxin with strong base, a fragment was isolated that contained bound, non-phosphorylated 3-deoxy-2-octulosonic acid, glucosamine phosphate and fatty acids. This indicated that polysaccharide I, like polysaccharide II, was bound to the lipid region of the endotoxin. The endotoxin structure thus defined is different from that proposed for the lipopolysaccharides of enterobacteria.