Jack Peter Green

Heparin, 5-hydroxytryptamine, and histamine in neoplastic mast cells.

Abstract During culture in vitro of an ascitic mast-cell tumor of the mouse, P-815, three strains have been developed, two of which were obtained by a cloning-technique. One of these pure strains is near diploid, the other tetraploid. All strains maintain their intracellular levels of heparin, histamine, and 5-hydroxytryptamine (5-HT) in culture. The levels of these three substances and the incorporation of S35-sulfate, glucosamine-1-C14, and glucose-U-C14 into heparin have been compared in the cells both in the mouse and in culture. Strains vary in their contents of these compounds and in their capacity to incorporate S35-sulfate and glucosamine-1-C14 into heparin, but in all cases the amounts of heparin, 5-HT, and histamine are higher in culture than when the same strain of cells are grown in the mouse; similarly, the incorporation of S35-sulfate into heparin is higher in culture. The amounts of the amines, especially of 5-HT, are proportional to the amount of heparin. The three substances are found in the same intracellular fraction. By paper chromatography mast-cell heparin has been resolved into three fractions. The concentration of one of these is proportional both to the amount of heparin, as determined by bioassay, and to the levels of amines, especially 5-HT. This component of heparin is not present in bovine heparin. It was also noted that sulfate and glucosamine in heparin turn over at the same rate. The half-life of heparin in the mast-cells is approximately two and one half days.

Binding of Some Biogenic Amines in Tissues

Publisher Summary This chapter discusses the binding of some biogenic amines in tissues. Almost all aspects of the physiology, pharmacology, and biochemistry of biogenic amines can be discussed from the point of view of their binding to tissues or tissue components. A binding mechanism has been implicated or demonstrated in (1) the biosynthesis, storage, transport, and release of amines, (2) their interaction with specific receptors or enzymes to elicit a response, and (3) their inactivation by enzymes and perhaps in their inactivation by binding to tissues. Clearly, each one of these processes merits separate review, but as information on the binding of biogenic amines is diffuse and the implications are broad, an eclectic approach seemed worthwhile. Therefore, this chapter encompasses binding in its widest sense, although no attempt is made to cover the biosynthesis of the amines or the catabolic enzymes. The same cells that contain biogenic amines almost invariably contain enzymes that inactivate the amines. An indication of the presence in tissues of a mechanism for storing endogenous amines is the finding that the histamine levels of tissues correlate with the capacities of the tissues to bind endogenously formed histamine and not with their histidine decarboxylase activities.

The uptake of amino acids and the synthesis of amines by neoplastic mast cells in culture.

For 4 years, two lines of neoplastic murine mast cells, both descendents of single cells, one a near-diploid, the other a near-tetraploid have been growing in culture in this laboratory. During this period they have retained their capacity to synthesize 5-hydroxytryptamine and heparin (Green & Day, 1960). It has been inferred that they also continue to synthesize histamine, for the immeasurably low histamine content of the culture medium (Schindler, Day & Fischer, 1959) cannot account for the persistent levels of this amine in the cells (Green & Day, 1960). In this paper some observations are presented on the uptake and metabolism by neoplastic mast cells of tryptophan, 5-hydroxytryptophan, histidine, and 3,4-dihydroxyphenylalanine. Preliminary reports of some of this work have been published (Day & Green, 1960; Green & Day, 1961).

Some acidic substances in neoplastic mast cells and in the pineal body

Abstract In neoplastic mast cells in culture the levels of histamine and 5-hydroxytryptamine reflected the levels of taurine, cysteic acid, cerebroside sulfate, and adenosine triphosphate and the capacity of the cells to synthesize heparin. No neuraminic acid could be detected in these cells. In the pineal body, high levels of neuraminic acid were found along with significant concentrations of taurine and cysteic acid, but no cerebroside sulfate or sulfomucopolysaccharides were detectable. Reserpine did not affect the levels of taurine in brain, spleen, or heart of the rat.

The uptake of biogenic amines by neoplastic mast cells in culture

Neoplastic mast cells ofthe mouse that have been continuously in culture in this laboratory for the past 4 years have maintained their ability to synthesize heparin (Green & Day, 1960), histamine and 5-hydroxytryptamine (Day & Green, 1962). The levels of amines in the cells fluctuate with time, whether the cells are grown in culture or as ascitic tumours in the mouse, and the levels appeared to correlate with the capacity of the cells to take up and decarboxylate the precursor amino acids, histidine and 5-hydroxytryptophan (5-HTP). This correlation was especially clear in a comparison of two lines of these cells, each a descendant of a single cell. The X-1 cells almost invariably showed higher levels of amines than did the X-2 cells, a difference that was reflected both in a more rapid uptake of histidine, 5-HTP, and tryptophan and in a more rapid catabolism of these amino acids to the amines. In this paper evidence is presented that these cells also take up pre-formed amines, and that X-2 cells exceed X-1 cells in this capacity. Preliminary reports of this work have appeared (Day & Green, 1960; Green & Day, 1961).

BIOSYNTHETIC PATHWAYS IN MASTOCYTOMA CELLS IN CULTURE AND IN VIVO

Mast cells, which have been detected in every species studied from sponges to primates, are in man found in connective tissue (Riley, 1959). Of the many roles that have been attributed to mast cells, least equivocal are their proposed functions in the allergic response and in the reaction to infections (Sheldon and Bauer, 1960; Wells, 1962) and to noxious chemicals ( Higginbotham, 1959). Provocative suggestions have been made that these cells play a role in diseases such as periarteritis nodosa, rheumatoid arthritis ( Smyth and Gum, 1961), nephrosclerosis ( Pavone-Macaluso, 1960), and atherosclerosis ( Sundberg, 1955; Pomerance, 1958; Pollack, 1957; Watson, 1961). Most of the inferences about the function of mast cells relate to their content of three physiologically active substances: heparin, histamine (Riley, 1959; West, 1959), and in the mouse and rat, 5-hydroxytryptamine ( Benditt, Wong, Arase, and Roeper, 1955; Parratt and West, 1957).

Effects of Liver Fractions on Myocardial Contractility

Tyramine was crystallized and identified as one of the cardioactive materials in liver extract. Of many other compounds tested on the isolated papillary muscle of the cat, only methionine, ethionine, serotonin, heparin, testosterone, Menadione, and Dicoumarol showed activity.

Blood 5-hydroxytryptamine (serotonin) levels after reserpine and electroshock therapy.

Summary Blood 5-HT fell to non-detectable levels after administration of reserpine to patients during a 3 week period. Three weeks after withdrawal of the drug, 5-HT levels had risen but had not yet reached normal values. Electro-shock therapy did not alter the levels of 5-HT in the blood.

ACETYLCHOLINE ACTIVITY IN THE SCIATIC NERVE.

Abstract As measured on both the guinea pig ileum and the frog rectus abdominis muscle, acetylcholine (Ach) is uniformly distributed along the sciatic nerve. Measured on the ileum, about 75 per cent of the Ach activity is found in the soluble portion of the cell, while on the rectus abdominis, the corresponding figure is about 35 per cent. Total Ach equivalents in sciatic nerve are higher when measured on the rectus abdominis muscle than on the ileum. Evidence is presented that the activity measured on the ileum is attributable to either Ach or a very similar substance. It is suggested that sciatic nerve contains a substance other than Ach to which the frog rectus abdominis muscle is especially sensitive.

The sulfomucopolysaccharides from a mast cell tumor of the mouse

Abstract Sulfomucopolysaccharides prepared from a murine mast cell tumor were fractionated by chromatography on Ecteola, by precipitation with cetyltrimethylammonium bromide, and by paper chromatography. The crude extract of murine sulfomucopolysaccharides, when extracted by the same procedure that was used to extract bovine Sulfomucopolysaccharides, was associated with nitrogen-rich material, unlike the comparable bovine preparation. The nitrogen-rich heparin-containing material, which was precipitable with 0.25 M NaCl and strongly adsorbed to Ecteola, was immobile upon paper chromatography. Examination of material prepared in the same way from bovine lung, guinea pig lung, a canine mastocytoma, and the lung and kidney of the rat, showed that only the tissues of the rat, like the murine mast cells, contained an immobile component. Purified murine sulfomucopolysaccharide showed a greater affinity for histamine and 5-hydroxytryptamine than did bovine sulfomucopolysaccharide. It is suggested that the relatively greater affinity of the murine sulfomucopolysaccharide for amino-containing compounds explains the high concentration of nitrogen in crude extracts of the murine sulfomucopolysaccharide.