A 57-kDa phosphatidylinositol-specific phospholipase C from bovine brain.

A phosphatidylinositol-specific phospholipase C (PI-PLC) has been isolated from bovine brain (purification factor of 5.6 x 10(4)). By sodium dodecyl sulfate-polyacrylamide gel electrophoresis, it had a Mr of 57,000. Neither amino nor neutral sugars were detected in the purified enzyme. The pH optimum was 7.0-7.5, and the activity decreased only slightly at pH 8.0. When phosphatidylinositol was used as a substrate, the optimum Ca2+ requirement was 4 mM, and Km was 260 microM. When phosphatidylinositol 4,5-bisphosphate was used, the optimum Ca2+ requirement was 10(-7) M, and the Km was reduced to 90 microM. Lipid specificity studies showed that equal amounts of inositol phosphate and diacylglycerol were released from phosphatidylinositol but 4 times as much inositol 1,4,5-trisphosphate was released from phosphatidylinositol 4,5-bisphosphate. Other lipids, phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin, were not substrates. Failure to detect phosphatidic acid confirmed the absence of a phospholipase D activity in the purified enzyme. Myelin basic protein (MBP) stimulated the PI-PLC activity between 2- and 3-fold. Histone had a small effect only, whereas bovine serum albumin and cytochrome C had no effect. Phosphorylation of MBP reduced the stimulatory effect. Protein-protein interactions between MBP and PI-PLC have been demonstrated both immunologically and by sucrose density gradients. A stoichiometry of 1:1 has been suggested by the latter method. A number of peptides have been prepared by chemical, enzymatic, and synthetic methods. Peptides containing the MBP sequences consisting of residues 24-33 and 114-122 stimulated the PI-PLC but were less effective than the intact protein.     

Spring feeding by pink-footed geese reduces carbon stocks and sink strength in tundra ecosystems

Tundra ecosystems are widely recognized as precious areas and globally important carbon (C) sinks, yet our understanding of potential threats to these habitats and their large soil C store is limited. Land‐use changes and conservation measures in temperate regions have led to a dramatic expansion of arctic‐breeding geese, making them important herbivores of high‐latitude systems. In field experiments conducted in high‐Arctic Spitsbergen, Svalbard, we demonstrate that a brief period of early season belowground foraging by pink‐footed geese is sufficient to strongly reduce C sink strength and soil C stocks of arctic tundra. Mechanisms are suggested whereby vegetation disruption due to repeated use of grubbed areas opens the soil organic layer to erosion and will thus lead to progressive C loss. Our study shows, for the first time, that increases in goose abundance through land‐use change and conservation measures in temperate climes can dramatically affect the C balance of arctic tundra.     

The use of the immobility reflex (animal hypnosis) as a possible procedure for detecting sedative activity

The immobility reflex was induced in rabbits and the amount of electrical current necessary to interrupt this state was determined before and after drug administration. Morphine and the major tranquilizers, chlorpromazine, haloperidol, and reserpine, elevated the arousal threshold over a wide dose range while the minor tranquilizers, chlordiazepoxide and meprobamate, were active over a considerably narrower range. The sedative-hypnotics, phenobarbital, pentobarbital, ethanol, chloral hydrate, and ectylurea demonstrated approximately the same degree of activity as the minor tranquilizers but resulted in a loss of righting reflex at the higher doses. D-Amphetamine significantly lowered the arousal threshold. Imipramine, desmethylimipramine, nialamide, aspirin, diphenylhydantoin, chlorpheniramine, and diphenhydramine possessed little, if any, activity. Thus, the drug-induced changes in arousal threshold in rabbits exhibiting the immobility reflex can be used to classify compounds possessing major or minor tranquilizing or sedative-hypnotic activity.     

Substance P evokes bradycardia by stimulation of postganglionic cholinergic neurons.

Substance P (SP) evokes bradycardia that is mediated by cholinergic neurons in experiments with isolated guinea pig hearts. This project investigates the negative chronotropic action of SP in vivo. Guinea pigs were anesthetized with urethane, vagotomized and artificially respired. Using this model, IV injection of SP (32 nmol/kg/50 microl saline) caused a brief decrease in heart rate (-30+/-3 beats/min from a baseline of 256+/-4 beats/min, n = 27) and a long-lasting decrease in blood pressure (-28+/-2 mmHg from baseline of 51+/-5 mmHg, n = 27). The negative chronotropic response to SP was attenuated by muscarinic receptor blockade with atropine (-29 +/- 9 beats/min before vs -8 +/- 2 beats/min after treatment, P = 0.0204, n = 5) and augmented by inhibition of cholinesterases with physostigmine (-23 +/- 6 beats/min before versus -74 +/- 20 beats/min after treatment, P = 0.0250, n = 5). Ganglion blockade with chlorisondamine did not diminish the negative chronotropic response to SP. In another series of experiments, animals were anesthetized with sodium pentobarbital or urethane and studied with or without vagotomy. Neither anesthetic nor vagotomy had a significant effect on the negative chronotropic response to SP (F3,24 = 1.97, P = 0.2198). Comparison of responses to 640 nmol/kg nitroprusside and 32 nmol/kg SP demonstrated that the bradycardic effect of SP occurs independent of vasodilation. These results suggest that SP can evoke bradycardia in vivo through stimulation of postganglionic cholinergic neurons.