Histochemical Demonstration of Dehydrogenase Activity in the Cells of Normal Human Blood and Bone Marrow

Abstract

Endogenous and succinic dehydrogenase activity was demonstrated in the living cells of normal human blood and bone marrow using a buffered nitro BT-succinate incubating solution. With this technique dchydrogenase activity was localized primarily in the granular leukocytes and the sites of cnzymatic activity appcared to be non-mitochondrial. The addition of a non-ionic surface active agent to thc incubating solution resulted in marked differences in the cellular and intracellular localization of dehydrogenase activity. With this method it was possible to demonstrate dehydrogenase activity in the mitochondria of most of the formed elements of the blood and bone marrow, including developing granulocytes and erythroid cells, agranulocytes, and blood platelets. Mature erythrocytes also exhibited a minimal dehydrogenase reaction with this procedure. This investigation indicated that in order adequately to demonstrate and evaluate dehydrogenase activity in the cells of the blood and bone marrow it was necessary to have increased cellular and mitochondrial permeability, as well as partially viable cells with an intact dehydrogenase system.

The Leukocytes and Platelets of a Marsupial, <i>Trichosurus vulpecula</i>

“…My finding that the specific granules are positive and resistant to amylase in Trichosurus parallels ACKERMAN's (1963) results with rabbit and guinea pig, and those of FEY (1966b) for some lower vertebrates.…”

“…By biochemical analysis, however, it has been found that only a small part of the PAS-positive content of leukocytes is glycogen (OLSSON and DAHLQVIST, 1967). It seems generally accepted that the glycogen of neutrophils occupies an intergranular position, though ACKERMAN (1964) says that much of it may be associated with the specific granules. Working with fractionated cells, VERCAUTEREN (1965) concluded that most was associated with the smooth endoplasmic reticulum though the specific granules may contain some.…”

“…ACKERMAN (1963) has also found the basophil granules to be negative to sudan black in rabbit and guinea pig. There have, on the other hand, been reports of positive sudan black staining of human basophlls (BLOOM and WISLOKI, 1950;WACHSTEIN, Lymphocytes.…”

“…As far as one can compare the basophil granules with those of other animals (some accounts in the literature are very sketchy and photographs are rarely presented) the granules in Trichosurus are of moderate size, relatively uniform size and shape, and relatively numerous and densely packed in the cytoplasm. In pig (SCARBOROUGH, 1931/2) the granules are plentiful and probably about the same size as in Trichosurus; in camels and llamas (PONDER, YEAGER and CHARIPPER, 1929a) and in cattle (DIMOCK and THOMPSON, 1906/71) they are large and numerous; in a number of Primates (PONDER, YEAGER and CHARIPPER, 1929b) they are coarse but the number varies with the species; in man himself (ACKERMAN, 1963;BLOOM and FAWCETT, 1968) they vary much in size and also in shape and, from my own observations, are less densely packed than in the cells of Trichosurus; in cat (SCARBOROUGH, 1931/2;GILMORE, GILMORE and JONES, 1964) the granules are large and sparse; in guinea pig (LUCIA and BLOOM and FAWCETT, 1968) (PONDER, YEAGER and CHARIPPER, 1929c), exhibits some basophilia in camels (PONDER, YEAGER and CHARIPPER, 1929a) and stains pink with blood stains in Macropus Lymphocytes. The nuclei of mammalian lymphocytes generally present a fairly standard appearance of a round or slightly indented outline with coarse and densely packed chromatin clumps, especially in the smaller cells.…”

“…In the neutrophils of cattle WINQVIST (1954) found some diffuse staining after amylase digestion, and SUGIYAMA (1955) found that the positive material was not amylase-labile in three non-mammalian species. After detailed investigations GAHRTON (1964) and MAHR (1964 have concluded that the PAS-positive material in human neutrophils is mainly glycogen; but ACKERMAN (1964) andSHUBICH (1965) mention some amylase-resistant material in the granules which the former considers to be a glycoprotein, the latter lipid. Similarly KAWABATA (1960), using isolated granules from fractionated horse leukocytes, found some stable PAS-positive material present in the granules.…”