The histamine and serotonin content of the platelets and polymorphonuclear leucocytes of various species

Humphrey, John; Jaques, R.

doi:10.1113/jphysiol.1954.sp005109

Cited by 200 publications

(63 citation statements)

References 12 publications

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“…The plasma was centrifuged to remove 5-HT and histamine contained in the platelets. The concentration of platelets remaining in the centrifuged plasma was apparently not high enough to influence the result through the release of 5-HT or histamine or an unidentified plain-muscle stimulating substance (Humphrey and Jaques, 1954).…”

Section: Discussionmentioning

confidence: 99%

A Method of Estimating Histamine in Plasma

Adam

Hardwick

Spencer

1957

British Journal of Pharmacology and Chemotherapy

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The method is intended for the estimation of minute quantities of histamine in blood plasma. A neutralized trichloracetic acid extract of plasma is adsorbed at pH 8 on a column prepared by mixing a quantity of the cationic exchange resin Amberlite XE-64 with powdered cellulose as a supporting medium. Histamine is adsorbed but N-acetylhistamine is not: the amines can therefore be estimated independently. Elution of the histamine is by displacement with HCL. The eluate is converted into the solution for bioassay on the superfused guinea-pig ileum.In recovery experiments histamine was added in the range 25 to 100 ng. to solutions of known composition and to plasma (5 ml.) obtained from man and the cat. The mean recovery in this range for histamine added to plasma was 82.5% A2 S.E. (17 estimations).The histamine equivalent of human plasma obtained from the antecubital vein was found to be less than 1 ng./ml.It was possible by this method to follow the concentration of histamine in the arterial plasma of the cat when histamine was infused intravenously at a rate of 330 ng./kg./min. There was no perceptible change in the arterial blood pressure during the infusion, but the plasma histamine rose from <0.3 ng./ml. to 3.4 ng./ml. (mean of 2 estimations).When the quantity of histamine extractable from plasma is less than can be estimated by conventional methods of bioassay, the sensitivity of the test can be increased by applying the undiluted extract directly to the strip of guinea-pig ileum; this may be suspended in a minute bath (Mongar and Schild, 1950) or in air, as in the technique of superfusion (Gaddum, 1953). The method is applicable if the histamine has been sufficiently purified and if the test solution is similar in composition to the solution bathing the intestine between the doses.If plasma is extracted by the method of Barsoum and Gaddum (1935) as modified by Code (1937a) and tested in this way, the activity of the extract is only partially abolished by the histamine antagonist mepyramine maleate (Mongar and Whelan, 1953). Moreover, the method does not distinguish between free histamine and histamine which may be released from pharmacologically inactive forms at the stage of refluxing the extract in strong acid (unpublished results).Various chromatographic methods of purifying histamine have been proposed and are reviewed in the article by Code and McIntire (1956). Some employ paper chromatography, others weak cationic exchangers which are used to remove histamine from solutions containing organic solvents.These methods are designed mainly for the chemical determination of histamine, but some have been combined with bioassay.In the method to be described the purification is conducted entirely in aqueous solution with the aid of a carboxylic ion exchange resin, at room temperature and at pH values not far from neutral. Advantage is taken of the fact that carboxylic resins have a high exchange capacity and can be buffered over a wide range of pH. They can therefore be used to separate weak organic bases in a...

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Section: Discussionmentioning

confidence: 99%

A Method of Estimating Histamine in Plasma

Adam

Hardwick

Spencer

1957

British Journal of Pharmacology and Chemotherapy

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“…Humphrey & Jaques (1954) have estimated the amounts of serotonin present in platelets of different species of animals, and have found that this substance is present in larger amounts than histamine. It is not known why platelets should contain such large amounts of serotonin.…”

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“…Red cells, free from platelets, were washed thrice with suspending fluid by centrifugation at 40 C. It had been observed in other experiments (Humphrey & Jaques, 1954) that rabbit platelets which were alternately frozen and thawed released into the surrounding fluid any serotonin or histamine which they contained. The amounts of histamine released in this way were, within experimental error, the same as could be extracted by brief * Singapore, Colonial Development and Welfare Scholar.…”

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Absorption of serotonin (5‐hydroxytryptamine) and histamine by dog platelets

Humphrey¹,

Toh²

1954

The Journal of Physiology

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115

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It is now well known that the vasoconstrictor substance in platelets is 5-hydroxytryptamine (serotonin) (Rand & Reid, 1951). Humphrey & Jaques (1954) have estimated the amounts of serotonin present in platelets of different species of animals, and have found that this substance is present in larger amounts than histamine. It is not known why platelets should contain such large amounts of serotonin. In this paper it will be shown that platelets from dogs are capable of taking up serotonin from solution. This finding raises the possibility that the serotonin found in platelets may be accumulated rather than formed in situ. METHODS Dogs were anaesthetized with phenobarbitone. A short glass cannula, previously treated with silicone, was inserted into the femoral artery and joined to a short length of polythene tubing for collection of blood.Preparation of platelets. Blood was collected into silicone-coated vessels and mixed with 7&th vol. of 1 % 'Versene' (disodium ethylenediaminetetra-acetic acid) in 0 7 % NaCl solution. After centrifugation for i hr at 1600 g in a refrigerated centrifuge, the buffy layer was collected and transferred to narrow silicone-coated centrifuge tubes, in which it was again centrifuged for J hr at 1600-2000 g. The top portion of the buffy layer could now quite easily be collected by means of a pipette coated inside and out with paraffin wax, and consisted almost entirely of platelets. There were few or no red cells, and the proportion of white cells to platelets was about 1:500-1:1000. The platelets were suspended in 10 ml. of the fluid described below, and centrifuged at 40 C for J hr at 2000 g. The supernatant (which contained negligible amounts of serotonin or histamine) was discarded and the washing repeated. The platelets were then suspended in the suspending fluid, and their number counted, using a haemocytometer. When stored at 40 C they were quite stable, showing no tendency to agglutinate or to lose their contained serotonin into the surrounding fluid over periods of 5 hr.Preparation of red cells: Lys8i ofplatelets. Red cells, free from platelets, were washed thrice with suspending fluid by centrifugation at 40 C. It had been observed in other experiments (Humphrey & Jaques, 1954) that rabbit platelets which were alternately frozen and thawed released into the surrounding fluid any serotonin or histamine which they contained. The amounts of histamine released in this way were, within experimental error, the same as could be extracted by brief * Singapore, Colonial Development and Welfare Scholar. Present address: Physiology Department, University of Malaya, Singapore.

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“…(b) The only other tissue elements known to contain 5HT in any significant concentration are platelets and mast cells. Available data (19) show that the quantity of platelets in any reasonable amount of residual blood is too small to contribute a significant amount of 5HT. Mast cells are not only rare in the guinea pig duodenum but also fail to release 5HT on treatment with reserpine (at least in the rat).…”

Section: Discussionmentioning

confidence: 99%

On the Concentration of 5-Hydroxytryptamine in Mammalian Enterochromaffin Cells and Its Release by Reserpine

Benditt

Wong

1957

The Journal of Experimental Medicine

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The site and local concentration of a chemical substance in the tissues are important determinants of its spatial interactions and hence its functional effects. 5-Hydroxytryptamine (5HT) has been identified in tissues rich in enterochroma/fin (E.C.) cells (1, 2) including carcinoid tumors (3). The histochemical reactions of these cells and of 5-hydroxytryptamine in models have been found to be similar (4). However, the negative reactions for indoles with p-dimethylaminobenzaldehyde in tissues containing enterochromaffin cells has recently been cited as evidence against the identity of enterochromarlin cell substance and 5-hydroxytryptamine (5).The following experiments were designed to test qualitatively and quantitatively the hypothesis that 5-hydroxytryptamine is the chromogenic material of enterochromaffin cells of the guinea pig duodenum. We shall produce evidence that 5-hydroxytryptamine is the chromogenic material of the enterochromaflfin cells, that the extractable 5-hydroxytryptamine is largely if not entirely present in these cells, and that the concentration in these cells is of the order of 1 per cent. It will be shown that reserpine causes the release of the 5-hydroxytryptamine from this cellular site. Methods and MaterialsAnimals.--For most of the experiments female guinea pigs weighing between 200 and 370 gin. were employed. The animals were housed two to four per cage. Market rabbits weighing 2.5 to 3.5 kg. were also used in one experiment. These were housed in individual cages. Animals were fed water and Rockland rabbit ration ad libitum.

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The histamine and serotonin content of the platelets and polymorphonuclear leucocytes of various species

Cited by 200 publications

References 12 publications

A Method of Estimating Histamine in Plasma

A Method of Estimating Histamine in Plasma

Absorption of serotonin (5‐hydroxytryptamine) and histamine by dog platelets

On the Concentration of 5-Hydroxytryptamine in Mammalian Enterochromaffin Cells and Its Release by Reserpine

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