The blood leukocytes of Bufo marinus: a light, phase-contrast, and histochemical study

1987 ◽  
Vol 65 (6) ◽  
pp. 1445-1453 ◽  
Author(s):  
M. Samuel Cannon ◽  
H. W. Sampson ◽  
E. D. Kapes
Keyword(s):  
Acid Phosphatase ◽  
Phase Contrast ◽  
Periodic Acid ◽  
Hydrolytic Enzymes ◽  
Neutral Red ◽  
Bufo Marinus ◽  
Oxidative Enzymes ◽  
Blood Leukocytes ◽  
Periodic Acid Schiff ◽  
Aerobic And Anaerobic

Blood leukocytes of Bufo marinus were studied by light and phase-contrast microscopy and histochemical techniques for the localization of glycogen, lipids, several basic proteins, and a number of hydrolytic and oxidative enzymes. The hydrolytic enzymes occurred in varying amounts in neutrophils, eosinophils, lymphocytes, and monocytes; neutrophils were the only leukocytes to demonstrate alkaline phosphatase activity, while β-glucuronidase was only seen in lymphocytes, and aryl-sulfatase was not observed in any leukocytes. Periodic acid – Schiff (PAS) positive granules also occurred in varying amounts in leukocytes. Slight lipid activity was only seen in neutrophils, while arginine, and (or) lysine, and tyrosine reactivity was only observed in eosinophils. The appearance and histochemical reactivity of acid phosphatase granules in neutrophils corresponded closely with the appearance and number of specific neutrophilic granules seen in Wright–Giemsa preparations and with the PAS-positive granules. Small lymphocytes were myeloperoxidase (peroxidase) negative; β-glucuronidase, acid phosphatase, and PAS-positive granules corresponded to neutral red granules seen in supravital films. The oxidative enzymes also occurred in differing amounts in leukocytes, but strongly suggested that the leukocytes of Bufo marinus are capable of some degree of aerobic and anaerobic metabolism.

The blood leukocytes of Bufo alvarius: a light, phase-contrast, and histochemical study

1979 ◽  
Vol 57 (2) ◽  
pp. 314-322 ◽  
Author(s):  
M. Samuel Cannon ◽  
A. M. Cannon
Keyword(s):  
Acid Phosphatase ◽  
Phase Contrast ◽  
Periodic Acid ◽  
Hydrolytic Enzymes ◽  
Neutral Red ◽  
Nonspecific Esterase ◽  
Alkaline Phosphatases ◽  
Blood Leukocytes ◽  
Periodic Acid Schiff ◽  
Aryl Sulfatase

Blood leukocytes of Bufo alvarius were studied by light and phase-contrast microscopy and histochemical techniques for the localization of glycogen and several hydrolytic enzymes, i.e., acid and alkaline phosphatases, nonspecific esterase, beta-glucuronidase, aryl-sulfatase, and myeloperoxidase (peroxidase). Neutrophils were the only leukocytes to demonstrate alkaline phosphatase activity, while beta-glucuronidase and aryl-sulfatase were not observed in any leukocytes. Periodic acid – Schiff (PAS) positive granules and granules containing hydrolytic enzymes occurred in varying amounts in leukocytes. In eosinophils, most glycogen was associated with smaller granules, while the larger refractile granules were PAS negative. Small lymphocytes were myeloperoxidase (peroxidase) negative. The present study agrees with previous investigations in mammals which indicate that specific granules in granulocytes may be PAS positive as well as contain one or more hydrolytic enzymes. In small lymphocytes of B. alvarius, PAS positive and acid phosphatase positive granules correspond to neutral red granules seen in supravital films. Furthermore, the appearance and histochemical reactivity of acid phosphatase granules in mature neutrophils, metamyelocytes, and late myelocytes correspond closely with the appearance and number of specific neutrophilic granules seen in Wright–Giemsa preparations and with PAS positive granules.

The morphology and cytochemistry of the blood leukocytes of Kemp's ridley sea turtle (Lepidochelys kempi)

1992 ◽  
Vol 70 (7) ◽  
pp. 1336-1340 ◽  
Author(s):  
M. Samuel Cannon
Keyword(s):  
Esterase Activity ◽  
Neutral Lipids ◽  
Periodic Acid ◽  
Sea Turtle ◽  
Oxidative Enzymes ◽  
Nonspecific Esterase ◽  
Blood Leukocytes ◽  
Periodic Acid Schiff ◽  
Bright Field Microscopy ◽  
Nonspecific Esterase Activity

Blood leukocytes of Lepidochelys kempi were examined by bright-field microscopy and cytochemistry for the determination of glycogen, lipids, and several hydrolytic and oxidative enzymes. Mature large and small eosinophils and small lymphocytes were the principle leukocytes encountered; basophils were rarely seen, and neutrophils and monocytes were not demonstrated. The large eosinophils contained two types of granules and demonstrated periodic acid – Schiff (PAS) reactivity and some sudanophilia. The large eosinophils also possessed alkaline phosphatase, myeloperoxidase, adenosine triphosphatase, and some nonspecific esterase activity. The small eosinophils demonstrated acid phosphatase. No reactivity for β-glucuronidase, aryl-sulfatase, or for the oxidative enzymes, succinate, lactate, and glucose-6-phosphate dehydrogenases, or cytochrome oxidase, occurred in the large or small eosinophils. Small lymphocytes contained a few PAS-positive granules or intracellular particles; some nonspecific esterase activity but no reactivity for other hydrolytic or oxidative enzymes or for neutral lipids was observed.

Prefertilization ovule development in Capsella: the dyad, tetrad, developing megaspore, and two-nucleate gametophyte

1986 ◽  
Vol 64 (4) ◽  
pp. 875-884 ◽  
Author(s):  
Patricia Schulz ◽  
William A. Jensen
Keyword(s):  
Cell Wall ◽  
Electron Microscope ◽  
Acid Phosphatase ◽  
De Novo ◽  
Periodic Acid ◽  
Aniline Blue ◽  
Ovule Development ◽  
Periodic Acid Schiff ◽  
Fluorescent Material ◽  
Autophagic Vacuoles

Ovules of Capsella bursa-pastoris at the dyad and tetrad stages of meiosis and at the megaspore and two-nucleate stages of the gametophyte were studied with the electron microscope. The cells of the dyad and tetrad are separated by aniline blue fluorescent cross walls and receive all types of organelles and autophagic vacuoles that were present in the meiocyte. Autophagic vacuoles enclose ribosomes and organelles and show reaction product for acid phosphatase. Autophagic vacuoles and some plastids are absorbed into the enlarging vacuoles of the growing megaspore. Other plastids appear to survive meiosis and there is no evidence for their de novo origin. Some mitochondria appear to degenerate in the enlarging megaspore but others look healthy and there is no evidence for the de novo origin of mitochondria. The nucleolus of the developing megaspore becomes very large and the cytoplasm is extremely dense with ribosomes. The cell wall is thickened by an electron-translucent, periodic acid – Schiff negative, aniline blue fluorescent material and contains plasmodesmata that link the megaspore with the nucellus. The plasmalemma of the growing megaspore produces microvilluslike extensions into this wall that disappear with the formation of the two-nucleate gametophyte. Plasmodesmata disappear from the cell wall at the four-nucleate stage.

Cellular differentiation and pattern formation in the absence of morphogenesis in the cellular slime mould Polysphondylium pallidum: evidence for a biochemical tip (organizer) in submerged aggregates

1981 ◽  
Vol 27 (9) ◽  
pp. 924-936 ◽  
Author(s):  
Gary D. Paterno ◽  
Danton H. O'Day
Keyword(s):  
Cell Differentiation ◽  
Cellular Differentiation ◽  
Periodic Acid ◽  
Neutral Red ◽  
Stalk Cell ◽  
Periodic Acid Schiff ◽  
Slime Mould ◽  
Polysphondylium Pallidum ◽  
Cellular Slime ◽  
Roller Tube Culture

When amoebae of Polysphondylium pallidum WS320 are placed in nonnutrient buffer in roller tube culture they form spherical or ellipsoidal aggregates. At first the aggregates demonstrate a "loose" morphology but by 12 h, with the formation of a cellulose-containing, peripheral sheath, they become "tight" aggregates. At this time stalk differentiation begins. Using various methods for the resolution of prespore (ultrastructure, spore antigen immunofluorescence, periodic acid – Schiff staining) and prestalk (ultrastructure, alkaline phosphatase histochemistry, neutral red staining, Calcofluor fluorescence) cell localization, the pattern of cell differentiation in submerged aggregates was shown to be essentially identical to that of normal pseudoplasmodia. Furthermore, using a cAMP bioassay it was revealed that the submerged aggregates, while devoid of a morphological tip, do possess a biochemical tip which is correlated with sites of neutral red staining and stalk cell differentiation. As a result of these studies, an earlier argument that the tip of the pseudoplasmodium is not essential for the establishment of pattern or in the "organization" of cellular differentiation during slime mould development is contradicted.

scholarly journals THE PARTICULATE HYDROLASES OF MACROPHAGES

1963 ◽  
Vol 118 (6) ◽  
pp. 1009-1020 ◽  
Author(s):  
Zanvil A. Cohn ◽  
Edith Wiener
Keyword(s):  
Acid Phosphatase ◽  
Yeast Cell ◽  
Cell Walls ◽  
Hydrolytic Enzymes ◽  
Neutral Red ◽  
Biochemical Properties ◽  
Extracellular Medium ◽  
Cell Content ◽  
Prolonged Incubation

The influence of phagocytosis on the morphological and biochemical properties of macrophage hydrolase-containing granules has been studied in vitro. Following the uptake of large numbers of heat-killed bacteria, an intracellular rearrangement of hydrolytic enzymes occurred. This was associated with the solubilization of 50 to 60 per cent of the total cell content of acid phosphatase, cathepsin, lysozyme, beta glucuronidase, acid ribonuclease, and acid desoxyribonuclease and with a corresponding decrease in granule-bound enzyme. With more prolonged incubation the majority of the soluble intracellular pool of acid ribonuclease and lysozyme was lost to the extracellular medium. No change in the total content of any of the hydrolases was noted during 180 minutes of incubation in vitro. The morphological fate of the granules was studied by a histochemical method for acid phosphatase. After the phagocytosis of yeast cell walls there was a disappearance of acid phosphatase-positive granules and an accumulation of reaction product about the ingested particle. Experiments employing macrophages which were supravitally stained with neutral red also demonstrated the loss of neutral red-positive granules and the accumulation of the dye about the yeast cell walls. These results strongly suggest that lysis of macrophage granules occurs following phagocytosis and that a portion of the granule contents are then resegregated within the newly formed phagocytic vacuole.

Acid phosphatase localization in PAS-bodies of Gonyaulax

1981 ◽  
Vol 51 (1) ◽  
pp. 15-23
Author(s):  
R.E. Schmitter ◽  
A.J. Jurkiewicz
Keyword(s):  
Acid Phosphatase ◽  
Phosphatase Activity ◽  
Acid Phosphatase Activity ◽  
Periodic Acid ◽  
Periodic Acid Schiff ◽  
Gonyaulax Polyedra ◽  
Marine Dinoflagellate ◽  
Pas Staining ◽  
First Time

Periodic acid-Schiff staining, acid phosphatase localization, and yellow autofluorescence have been correlated with the PAS-body of Gonyaulax polyedra for the first time. PAS- staining and acid phosphatase activity are both correlated with the PAS-body of Gonyaulax tamarensis. These results that the PAS-body of these marine dinoflagellate algae functions in subcellular digestion.

The Cytochemistry of the Amoebocytes and Intestinal Epithelium of Venus Mercenaria (Lamellibranchiata), with Remarks on a Pigment Resembling Ceroid

1955 ◽  
Vol s3-96 (33) ◽  
pp. 57-71
Author(s):  
SUMNER I. ZACKS
Keyword(s):  
Intestinal Epithelium ◽  
Periodic Acid ◽  
Goblet Cells ◽  
Neutral Red ◽  
Serum Cholinesterase ◽  
Periodic Acid Schiff ◽  
Intestinal Mucus ◽  
Specific Granules ◽  
Sulphydryl Groups ◽  
Metachromatic Granules

The properties of the amoebocytes and intestinal epithelium of Venus mercenaria were studied by a variety of cytochemical procedures designed to demonstrate proteins, enzymes, carbohydrates, and lipids. The cytoplasm of the amoebocytes contains specific granules which are constantly present and which are interpreted as being atypical mitochondria. Identification of their mitochondrial nature rests on their staining with Janus green B, their positive reaction for phospholipid by Baker's test, and the presence of dehydrogenase activity. Unlike typical mitochondria, the specific granules are eosinophil. Protein-bound carbonyl groups and disulphide and sulphydryl groups are present in both the specific granules and the cytoplasm. The sulphydryl groups may in part be associated with the presence of dehydrogenase, lipase, and serum cholinesterase. Amoebocytes also ontain glycogen and a material that is resistant to diastase and positive to the periodic acid / Schiff test; this material may be a neutral polysaccharide, unsaturated lipid, or mucoprotein. Cytoplasmic structures which are inconstantly present in amoebocytes include sudanophil droplets, neutral red vacuoles, metachromatic granules, and granules of an excretory pigment resembling ceroid. The sudanophil droplets may be stored neutral fat or lipid associated with the Golgi apparatus. The neutral red vacuoles are not preformed inclusions, but form as the dye accumulates within the cells. Metachromatic granules, which are confined solely to the intestinal amoebocytes, consist of phagocytosed intestinal mucus liberated from goblet cells. The histochemical reactions of the columnar intestinal epithelium suggest that these cells may be active in the digestion and absorption of nutrients, since eosinophil granules, lipid droplets, alkaline phosphatase, lipase, and serum cholinesterase are present in them. Masses of a ceroid-like excretory pigment and goblet cells containing mucus are present between the columnar intestinal epithelial cells. The pigment contains phospholipid and apparently arises as an oxidized end-product of lipid metabolism.

scholarly journals THE PARTICULATE HYDROLASES OF MACROPHAGES

1963 ◽  
Vol 118 (6) ◽  
pp. 991-1008 ◽  
Author(s):  
Zanvil A. Cohn ◽  
Edith Wiener
Keyword(s):  
Acid Phosphatase ◽  
Alveolar Macrophages ◽  
Intact Cell ◽  
Gradient Centrifugation ◽  
Hydrolytic Enzymes ◽  
Neutral Red ◽  
Mechanical Trauma ◽  
Freezing And Thawing ◽  
Alveolar Cells ◽  
Cell Content

The contents of selected hydrolytic enzymes of oil-induced peritoneal, normal alveolar, and BCG-induced alveolar macrophages have been studied. On a per cell or nitrogen basis the normal alveolar cells contained considerably more acid phosphatase, cathepsin, acid ribonuclease, lysozyme, and lipase than peritoneal cells. The BCG-induced alveolar macrophage exhibited increased levels of acid phosphatase, lysozyme, and lipase as compared to alveolar macrophages from unstimulated rabbits. The morphological differences between these cells was discussed and electron micrographs of the BCG-induced macrophage presented. Fractionation of the BCG-induced macrophage by differential centrifugation showed that 60 to 80 per cent of the total cell content of acid phosphatase, cathepsin, beta glucuronidase, acid ribonuclease, acid deoxyribonuclease, aryl sulfatase, lysozyme, and lipase were localized in a postnuclear fraction which sedimented at 15,000 g. This fraction also contained the majority of the mitochondria as evidenced by its content of cytochrome oxidase. Non-specific esterase was not localized to this fraction. A separation of the hydrolase-containing particles and mitochondria was achieved by isopycnic sucrose gradient centrifugation. Under the conditions employed, the mitochondria distributed at densities of 1.19 to 1.20, whereas the hydrolase particles sedimented to a density of 1.26 to 1.27. Each of the hydrolases including acid phosphatase, beta glucuronidase, cathepsin, lysozyme, and acid ribonuclease exhibited maximum activities in the same gradient fraction. The isolated granules exhibited enzymatic latency, and activation could be achieved by cycles of freezing and thawing or surface active agents. The majority of each of the hydrolytic enzymes could be liberated in a non-particulate form by mechanical trauma. Macrophages which had been stained supravitally with neutral red were fractionated by differential and gradient centrifugation. More than 70 per cent of the dye could be recovered in the particulate hydrolase fraction. The isolated, stained granules resembled those seen in the intact cell.

scholarly journals Structural and kinetic properties of a novel purple acid phosphatase from phosphate-starved tomato (Lycopersicon esculentum) cell cultures

2004 ◽  
Vol 377 (2) ◽  
pp. 419-428 ◽  
Author(s):  
Gale G. BOZZO ◽  
Kashchandra G. RAGHOTHAMA ◽  
William C. PLAXTON
Keyword(s):  
Acid Phosphatase ◽  
Lycopersicon Esculentum ◽  
Peptide Mapping ◽  
Periodic Acid ◽  
Purple Acid Phosphatase ◽  
Kinetic Properties ◽  
Suspension Cells ◽  
Periodic Acid Schiff ◽  
Α Subunit ◽  
Α And Β Subunits

An intracellular acid phosphatase (IAP) from Pi-starved (−Pi) tomato (Lycopersicon esculentum) suspension cells has been purified to homogeneity. IAP is a purple acid phosphatase (PAP), as the purified protein was violet in colour (λmax=546 nm) and was insensitive to l-tartrate. PAGE, periodic acid–Schiff staining and peptide mapping demonstrated that the enzyme exists as a 142 kDa heterodimer composed of an equivalent ratio of glycosylated and structurally dissimilar 63 (α-subunit) and 57 kDa (β-subunit) polypeptides. However, the nine N-terminal amino acids of the α- and β-subunits were identical, exhibiting similarity to the deduced N-terminal portions of several putative plant PAPs. Quantification of immunoblots probed with rabbit anti-(tomato acid phosphatase) immune serum revealed that the 4-fold increase in IAP activity due to Pi-deprivation was correlated with similar increases in the amount of antigenic IAP α- and β-subunits. IAP displayed optimal activity at pH 5.1, was activated 150% by 10 mM Mg2+, but was potently inhibited by Zn2+, Cu2+, Fe3+, molybdate, vanadate, fluoride and Pi. Although IAP demonstrated broad substrate selectivity, its specificity constant (Vmax/Km) with phosphoenolpyruvate was >250% greater than that obtained with any other substrate. IAP exhibited significant peroxidase activity, which was optimal at pH 9.0 and insensitive to Mg2+ or molybdate. This IAP is proposed to scavenge Pi from intracellular phosphate esters in −Pi tomato. A possible secondary IAP role in the metabolism of reactive oxygen species is discussed. IAP properties are compared with those of two extracellular PAP isoenzymes that are secreted into the medium of −Pi tomato cells [Bozzo, Raghothama and Plaxton (2002) Eur. J. Biochem. 269, 6278–6286].

66 HISTIOTROPHIC NUTRITION AND IRON TRANSFER ACROSS THE MATERNO-FETAL INTERFACE IN NUCLEAR TRANSFER-DERIVED SOMATIC CATTLE CLONES

2006 ◽  
Vol 18 (2) ◽  
pp. 141 ◽  
Author(s):  
F. Pereira ◽  
F. Meirelles ◽  
F. Braga ◽  
J. Visintin ◽  
R. Rumpf ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Periodic Acid ◽  
Cell Debris ◽  
Iron Transfer ◽  
Periodic Acid Schiff ◽  
Endometrial Stroma ◽  
Transporter Protein ◽  
Uterine Gland ◽  
Uterine Glands ◽  
Phosphatase Enzyme

The histiotrophic nutrition by the endometrial glands and the materno-fetal interface in the cloned cattle placenta were analyzed in order to investigate the iron transfer. Placentomes and intercaruncular region samples were recovered at term Caesarean delivery from 14 cloned cattle and 10 controls, fixed in 4% paraformaldehyde and 10% formaldehyde in PBS, processed and stained for light microscopy (hematoxylin-eosin, picrosirius, and Masson's trichrome), histochemistry [Perls, acid phosphatase and periodic acid Schiff (PAS) reactions], and immunohistochemistry (with rabbit anti-pig uteroferrin antibody because the uteroferrin is an iron transporter protein). In the controls we verified blood extravasations in the materno-fetal interface between the uterine and the trophoblast epithelium characterized by hemophagous areas with consequent erythrophagocytosis by the adjacent trophoblast. This content presented extravasated erythrocytes, plasm, cell debris, and cells in a probable apoptotic process. The Perls histochemical reactions that exposed the ferric iron in the placentomes were positive, as was the uteroferrin immunohistochemistry in the trophoblast cytoplasm and in other deep points in the placentomes. The histochemical reactions, demonstrating the acid phosphatase enzyme that detects the phagocytic activity, were positive in the mesenchyme and trophoblast, with a weak stain in an endometrial stroma. In the top of fetal villi, mainly in the binucleate cells, we visualized accumulations of PAS-positive secretions, indicating the presence of mucoid material. The uterine gland epithelium was columnar-type and in the gland lumina there were cell debris and PAS-positive mucoid secretions. We confirmed the reactivity of the uterine glands to the acid phosphatase enzyme and to the Perls reaction in the epithelium and in the gland lumina. The uteroferrin immunohistochemistry showed a strong stain in the cytoplasm of the endometrial glands cells and in the lumina. In the NT bovine placentae, the blood extravasations between uterine and trophoblast epithelium were aberrant. There was also the remodeling of the maternal connective tissue (endometrial stroma) in this area. We also demonstrated phagocytic uptake of uteroferrin by the trophoblast, although the histochemical and immunohistochemical reactions were weak in the trophoblast of the placentomes and in the endometrial glands of the intercaruncular region, when compared with the controls. The results obtained by the histochemistry and immunohistochemistry indicated that these sites of transfer substances from mother to fetus are very important in providing adequate nutrition to the fetus, key to a successful pregnancy in NT bovines. This work was funded by FAPESP, Brazil.

Sign in / Sign up

Export Citation Format

Share Document