Cytological and Cytochemical Studies on Cell Death and Digestion in the Foetal Rat Foot

1968 ◽  
Vol 3 (2) ◽  
pp. 245-262
Author(s):  
KATHLEEN J. BALLARD ◽  
S. J. HOLT
Keyword(s):  
Cell Death ◽  
Acid Phosphatase ◽  
Mesenchymal Cells ◽  
Enzyme Localization ◽  
Acid Hydrolases ◽  
Intracellular Release ◽  
Foetal Rat ◽  
Staining Methods ◽  
Hind Foot ◽  
Esterase Activities

Physiological cell death and degeneration in the interdigital mesenchyme of the hind foot of the rat foetus have been studied using classical staining methods and staining methods for enzyme localization. Individual mesenchymal cells die and shrink as the result of some unknown mechanism. Their acid phosphatase and esterase activities are not significantly different from those of viable loose mesenchymal cells. The dead cells are engulfed by viable neighbouring cells which resemble other loose mesenchymal cells in their morphology and in their acid phosphatase and esterase activities. These phagocytes then differentiate and become typical macrophages. Expressions of this process are the altered appearance of their nuclei and the increase in cytoplasm and in acid phosphatase and esterase activities. Many dead cells may be engulfed by a single macrophage and are then digested by its acid hydrolases. No evidence was found suggesting that cell death might be initiated by the intracellular release of lysosomal enzymes.

Cell death in the ‘opaque patch’ in the central mesenchyme of the developing chick limb: a cytological, cytochemical and electron microscopic analysis

Development ◽  
1971 ◽  
Vol 26 (3) ◽  
pp. 401-424
Author(s):  
D. S. Dawd ◽  
J. R. Hinchliffe
Keyword(s):  
Cell Death ◽  
Acid Phosphatase ◽  
Phosphatase Activity ◽  
Acid Phosphatase Activity ◽  
Mesenchymal Cells ◽  
Dead Cell ◽  
Acid Hydrolases ◽  
The Dead ◽  
Secondary Lysosomes ◽  
Mesenchyme Cells

Cell death in the ‘opaque patch’ of central mesenchyme of the developing chick forelimb was investigated by a variety of light and electron-microscope cytological and cytochemical techniques. Cell death appears first at stage 23/4 (4 days) and reaches its maximum extent at stages 24 and 25 (4½ and 5 days), at which it separates the ulnar and radial mesenchymal condensations. It then decreases in size to a small area separating the proximal parts of radius and ulna and disappears at stage 28. Cytological studies show the presence of a few isolated dead cells, of mesenchymal cells containing 1–3 ingested dead cells and of macrophages containing up to 18 dead cells in various stages of digestion. These findings are interpreted as showing that isolated dead cells are ingested by neighbouring mesenchymal cells which thus become transformed into macrophages, first ingesting and then digesting further dead cells. Histochemical studies show that isolated dead cells and recently ingested dead cells contain no more acid phosphatase activity, either discrete or diffuse, than either neighbouring living mesenchymal cells, or mesenchymal cells which have ingested 1–3 dead cells. Increased acid phosphatase activity is found within the macrophages, where activity is localized within the digestive vacuoles (‘secondary lysosomes’) containing the dead cells, and within the Golgi apparatus and Golgi vesicles (‘primary lysosomes’) of macrophage cytoplasm. Loss of staining capacity by the dead cell is correlated with high acid phosphatase activity: this is interpreted as indicating the digestion of dead cells within the macrophage by acid hydrolases. There is circumstantial evidence that viable mesenchyme cells in the ‘opaque patch’ area autophagocytose part of their own cytoplasm in secondary lysosomes (1·2–2µm). The role of the ‘opaque patch’ in relation to the pattern of limb chondrogenesis is discussed. It is suggested that cell death may play a role in separation of radius and ulna, and that autophagocytosis may indicate a change in the pathway of differentiation of the mesenchyme cells lying between radius and ulna.

vitreal cells and specialized phagocytes in the chick embryo retina: A TEM and SEM study

1990 ◽  
Vol 48 (3) ◽  
pp. 330-331
Author(s):  
M.A. Cuadros ◽  
M.J. Martinez-Guerrero ◽  
A. Rios
Keyword(s):  
Cell Death ◽  
Plasma Membrane ◽  
Acid Phosphatase ◽  
Chick Embryo ◽  
Basement Membrane ◽  
Sem Images ◽  
Intimate Contact ◽  
Cellular Processes ◽  
Sem Study ◽  
Free Cells

In the chick embryo retina (days 3-4 of incubation), coinciding with an increase in cell death, specialized phagocytes characterized by intense acid phosphatase activity have been described. In these preparations, all free cells in the vitreal humor (vitreal cells) were strongly labeled. Conventional TEM and SEM techniques were used to characterize them and attempt to determine their relationship with retinal phagocytes.Two types of vitreal cells were distinguished. The first are located at some distance from the basement membrane of the neuroepithelium, and are rounded, with numerous vacuoles and thin cytoplasmic prolongations. Images of exo- and or endocytosis were frequent; the cells showed a well-developed Golgi apparatus (Fig. 1) In SEM images, the cells was covered with short cellular processes (Fig. 3). Cells lying parallel to or alongside the basement membrane are elongated. The plasma membrane is frequently in intimate contact with the basement membrane. These cells have generally a large cytoplasmic expansion (Fig. 5).

Studies of Vegetative Plant Tissue Compatibility/Incompatibility: The Role of Acid Phosphatase in Lethal Cell Senescence in an Incompatible Heterograft

1980 ◽  
Vol 38 ◽  
pp. 530-531
Author(s):  
Randy Moore
Keyword(s):  
Acid Phosphatase ◽  
Thick Layer ◽  
Cell Necrosis ◽  
Enzyme Localization ◽  
Vegetative Plant ◽  
Cell Autolysis ◽  
Graft Interface ◽  
System 1 ◽  
Thin Fibril

Previous work has indicated that the graft incompatihility between Sedrmi telephoides and Solanum pennellil involves cell necrosis that results In a thick layer of collapsed cells at the graft Interface. This necrotic layer insulates the stock from the scion, which results in abscission of the Sedum scion after 4-6 weeks due to desiccation and starvation. Thus, cell autolysis (which is restricted to Sedum) characterizes the Incompatibility response in this system (1). In order to elucidate the events that lead to cell autolysis, and thus better understand the cellular site and mode of action of cellular incompatibility, the appearance and fate of the hydrolytlc enzyme acid phosphatase (AP) was followed in both the compatible Sedum autograft and the incompatible Sedum/Solanum heterograft. Acid phosphatase was localized by a modified Gomori-type reaction; positive (i.e., including NaF inhibitor) and negative (lacking substrate) controls showed no enzymatic precipitate. Following an initial association with the endoplasmic reticulum (ER) and dictyosomes at 6-10 hours after grafting, AP activity in the compatible Sedum autograft is associated primarily with the plasmalemma (Fig. 1). By 18-24 hours after grafting, the AP activity is restricted to the tono-plast and vacuole (Fig. 2). This strict compartmentation and absence of enzyme from the cytosol is maintained throughout the development of the compatible graft. While AP activity in the incompatible Sedum/Solanum heterograft is Initially similar to the compatible Sedum autograft (i.e., initially found on the ER and dictyosomes), there is a marked difference in enzyme localization in the two graft partners as the incompatibility response develops. As in the compatible autograft, Solanum cells at the graft interface show an Increase in AP activity that Is restricted to the vacuole and tonoplast, with little or no enzyme activity in the cytosol (Fig. 3). In comparable Sedum cells, however, there is a dramatic Increase In AP activity in the cytosol (Fig. h); this cytosollc AP activity is associated with thin fibril-like structures (Fig. 5) measuring approximately 60 A in diameter. This high cytoplasmic AP activity In Sedum cells results in cell autolysis, death, and eventual cell collapse to form the characteristic necrotic layer separating the two graft partners.

scholarly journals sFasL—The Key to a Riddle: Immune Responses in Aging Lung and Disease

2021 ◽  
Vol 22 (4) ◽  
pp. 2177
Author(s):  
Shulamit B. Wallach-Dayan ◽  
Dmytro Petukhov ◽  
Ronit Ahdut-HaCohen ◽  
Mark Richter-Dayan ◽  
Raphael Breuer
Keyword(s):  
Cell Death ◽  
Lung Disease ◽  
Fas Ligand ◽  
Death Receptor ◽  
Mesenchymal Cells ◽  
Soluble Form ◽  
Cell Accumulation ◽  
Key Factor ◽  
Aged Subjects

By dint of the aging population and further deepened with the Covid-19 pandemic, lung disease has turned out to be a major cause of worldwide morbidity and mortality. The condition is exacerbated when the immune system further attacks the healthy, rather than the diseased, tissue within the lung. Governed by unremittingly proliferating mesenchymal cells and increased collagen deposition, if inflammation persists, as frequently occurs in aging lungs, the tissue develops tumors and/or turns into scars (fibrosis), with limited regenerative capacity and organ failure. Fas ligand (FasL, a ligand of the Fas cell death receptor) is a key factor in the regulation of these processes. FasL is primarily found in two forms: full length (membrane, or mFasL) and cleaved (soluble, or sFasL). We and others found that T-cells expressing the mFasL retain autoimmune surveillance that controls mesenchymal, as well as tumor cell accumulation following an inflammatory response. However, mesenchymal cells from fibrotic lungs, tumor cells, or cells from immune-privileged sites, resist FasL+ T-cell-induced cell death. The mechanisms involved are a counterattack of immune cells by FasL, by releasing a soluble form of FasL that competes with the membrane version, and inhibits their cell death, promoting cell survival. This review focuses on understanding the previously unrecognized role of FasL, and in particular its soluble form, sFasL, in the serum of aged subjects, and its association with the evolution of lung disease, paving the way to new methods of diagnosis and treatment.

scholarly journals Tumor cell death visualization of renal cell carcinoma under the combined effect of the Gratiola officinalis extract and cyclophosphamide using fluorescent staining methods

2021 ◽  
pp. 2142004
Author(s):  
Artyom Mylnikov ◽  
Nikita Navolokin ◽  
Dmitry Mudrak ◽  
Natalya Polukonova ◽  
Alla Bucharskaya ◽  
...  
Keyword(s):  
Cell Death ◽  
Tumor Cells ◽  
Human Kidney ◽  
Combined Effect ◽  
Tumor Cell Death ◽  
Staining Methods ◽  
Apoptotic Activity ◽  
Apoptosis And Necrosis ◽  
Number Of Cells

Objective of the study: We used fluorescence imaging methods of apoptosis and necrosis in human renal carcinoma A498 tumor cells in vitro to reveal the indicated forms of cell death under the combined effect of flavonoid-containing extract of Gratiola officinalis and cytostatic (cyclophosphamide). Materials and methods: The dyes were propidium iodide and acridine orange, which were used in the “alive and dead” test. This test helped us to identify the total number of dead cells in the forms of necrosis and apoptosis and the number of cells in which apoptosis had started, it was characterized by the appearance of apoptotic bodies or nucleus pyknosis. Results: We found the most pronounced cytotoxic activity at the ratio of extract of Gratiola officinalis and cyclophosphamide concentrations of 1:1. The number of living cells decreased when exposed to the ratio of extract and cytostatic concentrations of 2:1. When the ratio of concentration of the extract relative to the cytostatic increased to 3:1, the cytostatic activity of the extract began to appear, the total number of tumor cells decreased. The number of cells with nucleus pyknosis and the number of cells with apoptosis signs significantly increased at a 3:1 ratio of extract and cytostatic concentrations, which confirms the presence of pro-apoptotic activity of the studied combination. This trend indicates the dependence of a certain form of cell death (apoptosis, necrosis) on the ratio of extract and cytostatic doses, and it also demonstrates the cytostatic and cytotoxic effects of this combination. Conclusion: Fluorescence methods of investigation in the “alive and dead” test allowed us to visualize the forms of cell death of human kidney carcinoma A498 by combined exposure to the flavonoid-containing extract of Gratiola officinalis and cytostatic (cyclophosphamide) 24 h after exposure. We found that the combination with a concentration ratio of the extract and cyclophosphamide of 3:1 has the greatest effectiveness due to stimulation of the cytostatic effect and cytotoxic effect.

Phosphatase Activity of Drosophila Salivary Glands

1948 ◽  
Vol s3-89 (8) ◽  
pp. 415-419
Author(s):  
W. L. DOYLE
Keyword(s):  
Acid Phosphatase ◽  
Enzymatic Activity ◽  
Quantitative Determination ◽  
Salivary Glands ◽  
Phosphatase Activity ◽  
Acid Phosphatase Activity ◽  
The State ◽  
Staining Methods

The phosphatases in the cytoplasm and nuclei of Drosophila salivary glands are better preserved by fixation in absolute acetone than in 85 per cent, alcohol. In whole glands there is relatively little extraction of the enzyme during assay. Phosphatase activity is more resistant to incubation at neutrality than at pH 8.6, but in this material there is sufficient residual enzymatic activity to permit redetermination of alkaline, neutral, or acid phosphatase activity by staining methods after an initial quantitative determination. The state of the membranes of the gland affects the penetration of the substrate sufficiently to limit the activities obtained.

scholarly journals Content and thrombin-induced release of acid hydrolases in gel-filtered platelets from patients with storage pool disease

Blood ◽  
1975 ◽  
Vol 46 (1) ◽  
pp. 131-142 ◽  
Author(s):  
H Holmsen ◽  
CA Setkowsky ◽  
B Lages ◽  
HJ Day ◽  
HJ Weiss ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Normal Subjects ◽  
Acid Hydrolases ◽  
Release Reaction ◽  
Dense Granules ◽  
Storage Pool ◽  
Low Concentrations ◽  
Storage Pool Disease ◽  
Beta Galactosidase

Abstract The levels of four acid hydrolases, beta-N-acetyl glucosaminidase, beta- glucuronidase, beta-galactosidase, and acid phosphatase, and the extent of their release (release II) by thrombin was determined in platelets from nine normal subjects, nine patients with storage pool disease, and in normal platelets which had been exposed to aspirin. The levels of all four hydrolases were normal in patients with SPD. However, release of three of these hydrolases (acid phosphatase was an exception) by low concentrations of thrombin (0.015 and 0.04 U/ml) was decreased in the patients as a group, although considerable variation in the extent of release of each enzyme was noted. In contrast, aspirin failed to inhibit release II in normal platelets (except for a slight impairment in the release of beta-N-acetyl glucosaminidase), although release I (serotonin, ATP and ADP) was inhibited. All release defects could be overcome by using higher concentrations of thrombin (0.2 U/ml). The normal levels of acid hydrolases in the platelets of patients with SPD (who are deficient in the platelet dense granules) suggest that these enzymes are not normally stored in the dense granules, but rather in alpha-granules. The findings also support the conclusions of previous studies that the release reaction is impaired in SPD. This release defect appears to be different from that seen in normal platelets after exposure to aspirin.

Metabolic events during programmed cell death in insect labial glands

1994 ◽  
Vol 72 (11-12) ◽  
pp. 597-601 ◽  
Author(s):  
Reginald Halaby ◽  
Zahra Zakeri ◽  
Richard A. Lockshin
Keyword(s):  
Cell Death ◽  
Protein Synthesis ◽  
Acid Phosphatase ◽  
Programmed Cell Death ◽  
Manduca Sexta ◽  
Adenosine Triphosphate ◽  
Mitochondrial Respiration ◽  
Second Messengers ◽  
Labial Gland ◽  
Dying Cells

The labial gland of Manduca sexta is a valuable system to study the mechanisms of programmed cell death since the death of the gland is nearly synchronous and, except for the anterior duct, involves all of the tissue. The gland degenerates in 5 days during pupation. Our previous work documents a drop in total protein synthesis as the gland degenerates. To evaluate potential causes of this altered protein synthesis, we monitored several parameters of metabolism in dying cells: levels of adenosine triphosphate to estimate the energy resources of the gland; reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide to assess mitochondrial respiration; levels of acid phosphatase to assay lysosomal enzyme activity; and concentrations of cyclic nucleotides and inositol triphosphate to monitor signaling. While protein synthesis fell precipitously on day 0, total adenosine triphosphate and mitochondrial respiration were unchanged until the cells underwent massive collapse on day 3. Lysosomal acid phosphatase increased during early metamorphosis, and ultimately the bulk of the cytoplasm was destroyed in autophagic vacuoles. Changes in the concentrations of second messengers were modest and late. The relationships between the metabolism and the collapse of the labial gland are under investigation.Key words: programmed cell death, Manduca sexta, energetics, lysosomes, second messengers, protein synthesis.

scholarly journals Distribution of starch, lipid and nuclei in xylem and phloem of Tectona grandis Linn.

2012 ◽  
Vol 19 ◽  
pp. 29-35 ◽  
Author(s):  
Md Azharul Islam ◽  
Shahanara Begum
Keyword(s):  
Cell Death ◽  
Tectona Grandis ◽  
Tree Breeding ◽  
Plant Tissues ◽  
Phloem Parenchyma ◽  
Tree Breeding Program ◽  
Teak Tree ◽  
Xylem Tissue ◽  
Staining Methods ◽  
Cell Death Pattern

Context: Reserve materials among different plant tissues vary species to species. The distribution pattern of such materials and cell death pattern in Tectona grandis Linn. are still obscure. Objectives: To study the localization of starch, lipid and nuclei in the phloem, cambium and xylem tissue of T. grandis. Materials and Methods: Blocks containing phloem, cambium and outermost xylem of the stem of 12 years old teak tree collected. Different staining methods used to visualize starch, lipid and nuclei within different cells under light microscope. Results: Starch in parenchyma cells is more abundant in outer xylem than phloem and cambium. Lipids droplets are uniformly distributed in outermost xylem. Phloem parenchyma content few mass of lipids but, limited in cambium. There are many dead cells visualized in both phloem and xylem with characteristic patterns. The results clarify the levels of starch, lipid in tissue of T. grandis and showed distinguished variation among the cell contents. Conclusion: The physiology of plant cells related to transport of nutrients and cell death also illustrated in the report. This would be helpful for further study to improve quality wood through tree breeding program. DOI: http://dx.doi.org/10.3329/jbs.v19i0.12997 J. bio-sci. 19: 29-35, 2011

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