Effects of Colchicine on the Golgi Complex and Gerl of Cultured Rat Peritoneal Macrophages and Epiphyseal Chondrocytes

1980 ◽  
Vol 45 (1) ◽  
pp. 41-58
Author(s):  
JOHAN THYBERG ◽  
ANDRZEJ PIASEK ◽  
STANISLAW MOSKALEWSKI
Keyword(s):  
Golgi Complex ◽  
Structural Integrity ◽  
Cell Types ◽  
Peritoneal Macrophages ◽  
Electron Microscopic ◽  
Thiamine Pyrophosphatase ◽  
Cytoplasmic Microtubules ◽  
The Individual ◽  
Functionally Diverse

Thioglycollate-elicited rat peritoneal macrophages and epiphyseal chondrocytes were cultured in vitro, treated with colchicine, and then studied by electron-microscopic and cytochemical techniques. Colchicine, but not lumicolchicine, caused disappearance of cytoplasmic microtubules and breakup of the Golgi complex with spreading of its dictyosomes from a well defined juxta nuclear area throughout the cytoplasm. There was also an altered distribution of lysosomes, which oriented themselves close to the dictyosomes both in controland colchicine-treated cells. Further, the structure of the individual dictyosomes was changed, especially in the chondrocytes. GERL equivalents were observed in control cells but were difficult to detect after exposure to colchicine. Reaction product for thiamine pyrophosphatase was found in narrow cisternae on the inner side of the dictyosomes in control cells but invacuole-like structures in colchicine-treated cells. Reaction product for acid phosphatase was present in GERL equivalents and lysosomes in control cells but mainly in lysosomes incolchicine-treated cells. Nevertheless, the total specific activities of these enzymes as well as of 5′-nucleotidase, a plasma membrane marker, remained unaffected by the drug treatment. These observations show that cytoplasmic microtubules play an important and, in many respects similar, cytoskeletal role in two so functionally diverse cell types as macrophages and chondrocytes. They are particularly important for the structural integrity of the Golgi complex, which in both cells is normally organized in the area around the centrioles, from which numerous microtubules radiate into the cytoplasm. The observations further suggest that GERL is an integrated part of the Golgi complex in these cells.

scholarly journals Proteome Profiling of PMJ2-R and Primary Peritoneal Macrophages

2021 ◽  
Vol 22 (12) ◽  
pp. 6323
Author(s):  
Alexander L. Rusanov ◽  
Peter M. Kozhin ◽  
Olga V. Tikhonova ◽  
Victor G. Zgoda ◽  
Dmitry S. Loginov ◽  
...  
Keyword(s):  
Proteomic Analysis ◽  
Living Organism ◽  
Peritoneal Macrophages ◽  
In Vitro Models ◽  
Individual Characteristics ◽  
Essential Proteins ◽  
Comparative Proteomic Analysis ◽  
Immortalized Cell ◽  
The Individual

In vitro models are often used for studying macrophage functions, including the process of phagocytosis. The application of primary macrophages has limitations associated with the individual characteristics of animals, which can lead to insufficient standardization and higher variability of the obtained results. Immortalized cell lines do not have these disadvantages, but their responses to various signals can differ from those of the living organism. In the present study, a comparative proteomic analysis of immortalized PMJ2-R cell line and primary peritoneal macrophages isolated from C57BL/6 mice was performed. A total of 4005 proteins were identified, of which 797 were quantified. Obtained results indicate significant differences in the abundances of many proteins, including essential proteins associated with the process of phagocytosis, such as Elmo1, Gsn, Hspa8, Itgb1, Ncf2, Rac2, Rack1, Sirpa, Sod1, C3, and Msr1. These findings indicate that outcomes of studies utilizing PMJ2-R cells as a model of peritoneal macrophages should be carefully validated. All MS data are deposited in ProteomeXchange with the identifier PXD022133.

Metabolism of 3′-Azido-3′-Deoxythymidine and 3′-Fluoro-3′-Deoxythymidine in Combination in Human Immunodeficiency Virus Infected Lymphoblastoid Cells

1992 ◽  
Vol 3 (3) ◽  
pp. 165-170 ◽  
Author(s):  
S. Cox
Keyword(s):  
Human Immunodeficiency Virus ◽  
Virus Replication ◽  
Cell Types ◽  
Human Immunodeficiency Virus Replication ◽  
Lymphoblastoid Cells ◽  
Synergistic Inhibition ◽  
Immunodeficiency Virus ◽  
Deoxynucleoside Triphosphate ◽  
The Individual

A combination of 3′-azido-3′-deoxythymidine (AZT) with 3′-fluoro-3′-deoxythymidine (FLT) has been shown previously to give synergistic inhibition of human immunodeficiency virus replication and greatly reduced cytotoxicity in vitro. The phosphorylation of the compounds, and their effect upon the natural deoxynucleoside triphosphate pools, were compared in CEM, H9, and HIV-infected H9 lymphoblastoid cells, both for the compounds when used alone and when combined together. Higher levels of FLT triphosphate than AZT triphosphate, and higher levels of AZT monophosphate than FLT monosphosphate, were formed in all cell types. Both compounds were phosphorylated most efficiently in CEM cells, whereas they were least efficiently phosphorylated in infected H9 cells. Owing to competition, the phosphorylation of both analogues was reduced when used in combination, compared to the phosphorylation of the separate compounds. The phosphorylation of the separate compounds was therefore at a maximum and was not increased by combining the compounds. The two compounds competed equally with each other for phosphorylation when used at a ratio of AZT: FLT of 5: 1. Both analogues severely reduced the deoxynucleoside triphosphate pools in uninfected and human immunodeficiency virus-infected H9 cells, but not in CEM cells. The effects of the two compounds were similar to those found when the compounds were combined, and thus H9 cells were shown to be much more sensitive to the effects of the analogues upon deoxynucleoside triphosphate pools than CEM cells were. Thus the synergistic combination of 3′-azido-3′-deoxythymidine and 3′-fluoro-3′-deoxythymidine was shown to have a similar metabolism and a similar effect upon cellular deoxynucleoside triphosphate pools to the individual compounds.

scholarly journals Golgi spectrin: identification of an erythroid beta-spectrin homolog associated with the Golgi complex.

1994 ◽  
Vol 127 (3) ◽  
pp. 707-723 ◽  
Author(s):  
K A Beck ◽  
J A Buchanan ◽  
V Malhotra ◽  
W J Nelson
Keyword(s):  
Golgi Complex ◽  
Structural Integrity ◽  
Functional Organization ◽  
Brefeldin A ◽  
Cell Types ◽  
Loss Of Function ◽  
Dynamic Relationship ◽  
Golgi Membranes ◽  
Golgi Structure ◽  
And Function

Spectrin is a major component of a membrane-associated cytoskeleton involved in the maintenance of membrane structural integrity and the generation of functionally distinct membrane protein domains. Here, we show that a homolog of erythrocyte beta-spectrin (beta I sigma*) co-localizes with markers of the Golgi complex in a variety of cell types, and that microinjected beta-spectrin codistributes with elements of the Golgi complex. Significantly, we show a dynamic relationship between beta-spectrin and the structural and functional organization of the Golgi complex. Disruption of both Golgi structure and function, either in mitotic cells or following addition of brefeldin A, is accompanied by loss of beta-spectrin from Golgi membranes and dispersal in the cytoplasm. In contrast, perturbation of Golgi structure without a loss of function, by the addition of nocodazole, results in retention of beta-spectrin with the dispersed Golgi elements. These results indicate that the association of beta-spectrin with Golgi membranes is coupled to Golgi organization and function.

scholarly journals Effect of molecular size of 125I-labelled poly(vinylpyrrolidone) on its pinocytosis by rat visceral yolk sacs and rat peritoneal macrophages

1981 ◽  
Vol 196 (1) ◽  
pp. 49-55 ◽  
Author(s):  
R Duncan ◽  
M K Pratten ◽  
H C Cable ◽  
H Ringsdorf ◽  
J B Lloyd
Keyword(s):  
Molecular Weight ◽  
Molecular Size ◽  
Weight Fraction ◽  
Cell Types ◽  
Peritoneal Macrophages ◽  
Yolk Sac ◽  
High Molecular Weight Fraction ◽  
Molecular Weights ◽  
Molecular Weight Distributions

Rates of pinocytosis of different molecular-weight distributions of 125I-labelled poly(vinylpyrrolidone) by rat visceral yolk sacs and rat peritoneal macrophages were measured in vitro. Four preparations of mean molecular weights 50 000, 84 000, 700 000 and 7 000 000, were used. Macrophages captured the highest-molecular-weight preparation more rapidly than the other preparations. In contrast, rate of capture by the yolk sac decreased with increasing molecular weight. Incubations with a very-high-molecular-weight fraction derived from the 7 000 000-average-mol. wt. preparation clearly demonstrated that very large polymer molecules are not accumulated by the yolk sac, but are preferentially captured by macrophages. Analysis of the 125I-labelled poly(vinylpyrrolidone) internalized by the two cell types confirmed that low-molecular-weight material is preferred by the yolk sac, whereas the macrophage is less discriminating.

scholarly journals The beads in the Golgi complex-endoplasmic reticulum region.

1976 ◽  
Vol 70 (2) ◽  
pp. 384-394 ◽  
Author(s):  
M Locke ◽  
P Huie
Keyword(s):  
Endoplasmic Reticulum ◽  
Rough Endoplasmic Reticulum ◽  
Golgi Complex ◽  
Insect Cell ◽  
Cell Types ◽  
Feulgen Staining ◽  
Clear Halo ◽  
Bismuth Salts ◽  
Transition Vesicles

The region between the rough endoplasmic reticulum (ER) and the Golgi complex has been studied in a variety of insect cell types in an attempt to find a marker for the exit gate or gates from the ER. We have found that the smooth surface of the rough endoplasmic reticulum near Golgi complex transitional elements has beadlike structures arranged in rings at the base of transition vesicles. They occur in all insect cell types and a variety of other organisms. The beads can be seen only after staining in bismuth salts. They are 10-12 nm in diameter and are separated from the membrane and one another by a clear halo giving them a center to center spacing of about 27 nm. The beads are not sensitive to nucleases under conditions which disrupt ribosomes or remove all Feulgen staining material from the nucleus. Under conditions similar to those used to stain tissue, bismuth does not react in vitro with nucleic acids. The component of the beads that stains preferentially with bismuth is therefore probably not nucleic acid.

scholarly journals A Review on Animals Semen Characteristics: Fertility, Reproduction and Development

2019 ◽  
pp. 1-9
Author(s):  
Farzad Moradpour
Keyword(s):  
Structural Integrity ◽  
Current Knowledge ◽  
Semen Analysis ◽  
Kinetic Property ◽  
Genetic Material ◽  
Computer Assisted ◽  
Semen Characteristics ◽  
The Individual

In this research, the goal of review was summarizing the current knowledge of the methods available to assess in vitro quality of frozen-thawed bovine spermatozoa also, a review on animal’s semen characteristics: fertility, reproduction and development after AI with that semen. Artificial insemination (AI) is the first generation reproductive biotechnology that has made a deep contribution to the genetics improvement in several animals. A fertile ejaculate must meet certain semen characteristics quality standards, such as: normal morphology, active energy metabolism, progressive motility, structural integrity and functionality of the membrane, penetration capacity and optimum transfer of genetic material. The percentage of total motile spermatozoa in normal canine ejaculates is between 70 to 90%. By the way, there are a lot of parameters that able to change on the composition and structure of various sperm plasma member domains, such as change temperature and sensitive to any theirs environments in vivo and vitro (tropical climates), season also nutrition. Computer-assisted semen analysis (CASA) is primarily used to obtain accurate and objective kinetic sperm measurements that gives extensive information about the kinetic property of the ejaculate based on measurements of the individual sperm cells.

scholarly journals Human eosinophil hematopoiesis studied in vitro by means of murine eosinophil differentiation factor (IL5): production of functionally active eosinophils from normal human bone marrow

Blood ◽  
1988 ◽  
Vol 71 (3) ◽  
pp. 646-651
Author(s):  
EJ Clutterbuck ◽  
CJ Sanderson
Keyword(s):  
Bone Marrow ◽  
Cell Types ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Electron Microscopic ◽  
Human Eosinophil ◽  
Interleukin 5 ◽  
Differentiation Factor ◽  
Microscopic Features

The production of human eosinophils in vitro from normal bone marrow by using murine eosinophil differentiation factor (mEDF/interleukin 5) is described. Eosinophil production was selective and first detectable after 14 days and reached a peak between 21 and 35 days when they were the predominant cell type (41% to 89%). Until day 14, all the eosinophils were typical myelocytes, developing thereafter into metamyelocytes and mature cells. All cell types had characteristic light- and electron-microscopic features, apart from the absence of granules with crystalline cores. The eosinophils produced were readily recovered, and both immature myelocytes and mature cells were functionally active in an antibody-dependent, cell-mediated cytotoxicity assay. mEDF added into the assay enhanced the cytotoxicity but to a lower degree than previously reported for peripheral blood eosinophils, which suggests that they may be partially activated. The possibility that eosinophils could be deactivated was tested by removing mEDF from the culture medium. The eosinophils retained viability and functional activity, however, and showed no increased ability to be activated by mEDF for up to six days after removing the mEDF. The liquid culture of human bone marrow was shown to be an alternative assay for eosinophil differentiation factors to colony formation.

scholarly journals Human eosinophil hematopoiesis studied in vitro by means of murine eosinophil differentiation factor (IL5): production of functionally active eosinophils from normal human bone marrow

Blood ◽  
1988 ◽  
Vol 71 (3) ◽  
pp. 646-651 ◽  
Author(s):  
EJ Clutterbuck ◽  
CJ Sanderson
Keyword(s):  
Bone Marrow ◽  
Cell Types ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Electron Microscopic ◽  
Human Eosinophil ◽  
Interleukin 5 ◽  
Differentiation Factor ◽  
Microscopic Features

Abstract The production of human eosinophils in vitro from normal bone marrow by using murine eosinophil differentiation factor (mEDF/interleukin 5) is described. Eosinophil production was selective and first detectable after 14 days and reached a peak between 21 and 35 days when they were the predominant cell type (41% to 89%). Until day 14, all the eosinophils were typical myelocytes, developing thereafter into metamyelocytes and mature cells. All cell types had characteristic light- and electron-microscopic features, apart from the absence of granules with crystalline cores. The eosinophils produced were readily recovered, and both immature myelocytes and mature cells were functionally active in an antibody-dependent, cell-mediated cytotoxicity assay. mEDF added into the assay enhanced the cytotoxicity but to a lower degree than previously reported for peripheral blood eosinophils, which suggests that they may be partially activated. The possibility that eosinophils could be deactivated was tested by removing mEDF from the culture medium. The eosinophils retained viability and functional activity, however, and showed no increased ability to be activated by mEDF for up to six days after removing the mEDF. The liquid culture of human bone marrow was shown to be an alternative assay for eosinophil differentiation factors to colony formation.

scholarly journals An Interpenetrating Alginate/Gelatin Network for Three-Dimensional (3D) Cell Cultures and Organ Bioprinting

Molecules ◽  
2020 ◽  
Vol 25 (3) ◽  
pp. 756 ◽  
Author(s):  
Qiuhong Chen ◽  
Xiaohong Tian ◽  
Jun Fan ◽  
Hao Tong ◽  
Qiang Ao ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Structural Integrity ◽  
Polymer Network ◽  
Three Dimensional ◽  
Biochemical Properties ◽  
Interpenetrating Polymer Network ◽  
3D Cell Cultures ◽  
Wide Range ◽  
The Individual

Crosslinking is an effective way to improve the physiochemical and biochemical properties of hydrogels. In this study, we describe an interpenetrating polymer network (IPN) of alginate/gelatin hydrogels (i.e., A-G-IPN) in which cells can be encapsulated for in vitro three-dimensional (3D) cultures and organ bioprinting. A double crosslinking model, i.e., using Ca2+ to crosslink alginate molecules and transglutaminase (TG) to crosslink gelatin molecules, is exploited to improve the physiochemical, such as water holding capacity, hardness and structural integrity, and biochemical properties, such as cytocompatibility, of the alginate/gelatin hydrogels. For the sake of convenience, the individual ionic (i.e., only treatment with Ca2+) or enzymatic (i.e., only treatment with TG) crosslinked alginate/gelatin hydrogels are referred as alginate-semi-IPN (i.e., A-semi-IPN) or gelatin-semi-IPN (i.e., G-semi-IPN), respectively. Tunable physiochemical and biochemical properties of the hydrogels have been obtained by changing the crosslinking sequences and polymer concentrations. Cytocompatibilities of the obtained hydrogels are evaluated through in vitro 3D cell cultures and bioprinting. The double crosslinked A-G-IPN hydrogel is a promising candidate for a wide range of biomedical applications, including bioartificial organ manufacturing, high-throughput drug screening, and pathological mechanism analyses.

Propagation of JC Virus in Glia Cells in PML (Progressive Multifocal Leukoencephalopathy

1975 ◽  
Vol 33 ◽  
pp. 328-329
Author(s):  
S. Preskorn ◽  
J. Kepes ◽  
W.J. Bopp ◽  
I. Watanabe
Keyword(s):  
Demyelinating Disease ◽  
Jc Virus ◽  
Cell Types ◽  
Autopsy Material ◽  
Electron Microscopic ◽  
Biopsy Material ◽  
Papova Virus ◽  
Viral Particles ◽  
New Type ◽  
Cytoplasmic Microtubules

PML, a rare form of subacute demyelinating disease of the human brain, is caused, in most instances, by JC virus, a new type of papova virus. Light microscopically, the lesions are characterized by hypertrophy of oligodendroglia with intranuclear inclusion and gigantic astroglia with bizarreshaped nuclei. Although viral particles have been found by many investigators, the use of autopsy material has limited the ultrastructural study of changes in the patient.This report concerns some new cytopathological changes based on an electron microscopic study of well fixed biopsy material. Oligodendroglia and astroglia were markedly hypertrophic, but were identified by the presence of abundant cytoplasmic microtubules and filaments, respectively. Intranuclear and intracytoplasmic virions were found in both cell types. The intranuclear virions were the typical round and filamentous forms, with diameters of 40 and 30 mμ, respectively. They were scattered randomly throughout the enlarged nuclei.

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