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 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 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 HYDROLYTIC ENZYMES DURING SPERMATOGENESIS IN RAT AN ELECTRON MICROSCOPIC AND HISTOCHEMICAL STUDY

1968 ◽  
Vol 16 (4) ◽  
pp. 249-262 ◽  
Author(s):  
ZOLTAN POSALAKI ◽  
DEZSÖ SZABÓ ◽  
ERNÖ BÁCSI ◽  
ISTVÁN ÖKRÖS
Keyword(s):  
Endoplasmic Reticulum ◽  
Enzyme Activity ◽  
Acid Phosphatase ◽  
Sertoli Cells ◽  
Hydrolytic Enzymes ◽  
Appreciable Amount ◽  
Second Phase ◽  
Nonspecific Esterase ◽  
Electron Microscopic ◽  
Two Phases

The localization of lipids and the activities of nonspecific esterase, aryl sulfatase and acid phosphatase were studied in different stages of spermatogenesis in rats. In addition, the distribution of acid phosphatase activity was demonstrated electron histochemically. The spermatogenetic cycle was divided into two phases—corresponding to the first and the last four stages of Roosen-Runge-Giesel (RG) classification. Spermatids in the first phase contained abundant endoplasmic reticulum with rosette formation and well developed Golgi apparatus with numerous vesicles. They displayed high activity of hydrolytic enzymes but contained no appreciable amount of lipids. The Sertoli cells contained large lipid granules but showed minimal enzyme activity. During the second phase reduction of the cytoplasm of spermatids with fragmentation of the endoplasmic reticulum and Golgi lamellae, accumulation of lipids, aggregation of ribonucleo-protein particles, formation of residual bodies and marked decrease of enzyme activity were seen. The Sertoli cells contained large mitochondria, well developed endoplasmic reticulum and numerous dense bodies and revealed high activities of hydrolytic enzymes and rapid depletion of lipids. These ultrastructural and histochemical findings suggested an interaction between the Sertoli cells and the developing spermatids which probably contributed to the regulation of spermatogenesis.

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.

scholarly journals STUDIES OF ACID PHOSPHATASE AND NONSPECIFIC ESTERASE ACTIVITIES IN RAT ADRENAL GLANDS FOLLOWING OPERATIVE STRESS

1970 ◽  
Vol 18 (9) ◽  
pp. 650-659 ◽  
Author(s):  
S. R. CHOUDHURY ◽  
A. M. LUNDY
Keyword(s):  
Acid Phosphatase ◽  
Esterase Activity ◽  
Adrenal Glands ◽  
Surgical Stress ◽  
Cauda Equina ◽  
Hydrolytic Enzymes ◽  
Nonspecific Esterase ◽  
Starch Gel Electrophoresis ◽  
Operative Stress ◽  
Esterase Activities

Acid phosphatase and esterase activities were studied in adrenal glands obtained from rats killed at regular intervals following surgical stress (cauda equina transection). Zymograms of acid phosphatase produced by starch gel electrophoresis revealed increased reactivity in the operated samples. With esterases, a slightly different pattern was observed in the operated group, which exhibited a few additional bands particularly in the cathode region. This was confirmed by densitometric analysis of the gel strips. Two of these additional bands were organophosphate-sensitive and the remaining few were activated by p-chloromercuribenzoate. These latter bands appeared to arise from splitting of the preexisting organophosphate-resistant bands present in control zymograms. Biochemical assay of the two hydrolytic enzymes demonstrated a remarkable similarity in their responses to operative stress—probably implying a general lysosomal activation. Both enzymes exhibited a peak activity 8 hr after operation, followed by a gradual decline. Both organophosphate-sensitive and organophosphate-resistant esterases contributed toward the rise in total esterase activity. Histochemical studies on tissue sections revealed a more reactive adrenal cortex in the operated group, but were of little help in localizing the additional esterase activity observed in gel strips.

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.

Demonstration of activated macrophages in subgingival plaque and crevicular fluid samples from periodontal diseased sites

1989 ◽  
Vol 47 ◽  
pp. 1084-1085 ◽  
Author(s):  
J. Hanker ◽  
E.J. Burkes ◽  
G. Greco ◽  
R. Scruggs ◽  
E. Anderson ◽  
...  
Keyword(s):  
Periodic Acid ◽  
Nonspecific Esterase ◽  
Subgingival Plaque ◽  
Activated Macrophages ◽  
Periodic Acid Schiff ◽  
Intense Staining ◽  
Gram Negative ◽  
Periodontal Tissues ◽  
Crevicular Fluid

Macrophages are present in gingival tissues and they increase in gingivitis and periodontitis. In vitro studies have shown that gram-negative bacterial endotoxin increases the bone resorption activity of macrophages. The activation of macrophages in vitro by substances such as lymphokines, complement, group A streptococcal wall substances, histamine and immune complexes which are present in subgingival plaque or periodontal tissues has also been shown. Intense staining of activated macrophages for acid phosphatase and nonspecific esterase has been observed in tissue sections of inflamed gingiva (cf. 4,9). Although macrophages have also been observed in crevicular fluid and subgingival plaques from normal sites, their numbers, enzyme activities or activation have not previously been studied in crevicular fluid and subgingival plaques from diseased sites. All of the following studies were performed on samples obtained by noninvasive procedures and smeared on coverslips. After cytochemical procedures which produced visible and electron opaque stains on replicate coverslips, correlative light and scanning electron microscopy by SEI and BEI modes permitted identification of cell types and some bacterial pathogens. The PATS reaction, a variation of the periodic acid-Schiff(PAS) reaction that employs stepwise condensation with thiocarbohydrazide and silver methenamine to achieve staining and electron opacity, has been found useful by us for positively staining gram-negative bacteria including spirochetes in crevicular fluid and subgingival plaques from periodontal patients. Neutrophils were intensely stained due to their glycogen content. Larger glycogen positive cells, frequently highly vacuolated, were also present; they appeared to be activated macrophages (Figs. 1-3).

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