scholarly journals LOCALIZATION OF A PRIMATE-SPECIFIC ESTERASE USING IMMUNOFLUORESCENCE AND IMMUNOPEROXIDASE TECHNIQUES

1973 ◽  
Vol 21 (6) ◽  
pp. 559-567 ◽  
Author(s):  
GEORGIRENE D. VLADUTIU ◽  
PIERLUIGI E. BIGAZZI ◽  
NOEL R. ROSE
Keyword(s):  
Cultured Cells ◽  
Monkey Kidney ◽  
Close Apposition ◽  
Proximal Tubule Cells ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Golgi Region ◽  
Collecting Tubules ◽  
Convoluted Tubules ◽  
Bowman's Capsule

The techniques of mixed antibody immunoperoxidase and indirect immunofluorescence were used for the localization of an esterase isoenzyme in human and monkey tissue culture cells and in monkey kidney frozen sections. In cultured cells, the esterase was found to be cytoplasmic in close apposition to the nucleus with a distribution resembling that of the Golgi region. The esterase in monkey kidney was predominantly found in the cytoplasm of the proximal convoluted tubules. There was slight staining in the Bowman's capsule of the glomerulus and in the collecting tubules of the medulla. It is suggested that the kidney is actually producing the esterase in its proximal tubule cells.

Formation of poliovirus in monkey kidney tissue culture cells

Virology ◽  
1962 ◽  
Vol 16 (3) ◽  
pp. 325-333 ◽  
Author(s):  
Heather Donald Mayor ◽  
Liane E. Jordan
Keyword(s):  
Tissue Culture ◽  
Kidney Tissue ◽  
Monkey Kidney ◽  
Culture Cells ◽  
Tissue Culture Cells

Analysis of Lysophagic Flux in Cultured Cells using Lyso-Keima v1

2021 ◽  
Author(s):  
Vinay V. Eapen ◽  
Sharan Swarup ◽  
Melisa Hoyer ◽  
Harper not provided JW
Keyword(s):  
Tissue Culture ◽  
Cultured Cells ◽  
Confocal Imaging ◽  
Signaling Proteins ◽  
Membrane Rupture ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Luminal Side ◽  
Galectin 3 ◽  
Autophagy Pathway

Lysophagy-the selective elimination of damaged lysosomes by the autophagy pathway-is a critical housekeeping mechanism in cells. This pathway surveils lysosomes and selectively demarcates terminally damaged lysosomes for elimination. Among the most upstream signaling proteins in this pathway are the glycan binding proteins-Galectins-which recognize N and O linked glycan chains on the luminal side of transmembrane lysosomal proteins. These glycosyl modifications are only accessible to galectin proteins upon extensive lysosomal membrane rupture and serve as a sensitive measure of lysosomal damage and eventual clearance by selective autophagy. Indeed, prior work has shown that immunofluorescence of Galectin-3 serves as a convenient proxy for lysophagic flux in tissue culture cells (Aits et al., 2015; Maejima et al., 2013). Here we describe a facile method for monitoring lysophagy using the acid sensitive fluorophore mKeima, affixed onto Galectin-3, which allows for the monitoring of lysophagic flux by Flow cytometry, Western blotting or Confocal imaging. This method, which we have termed Lyso-Keima, serves as a facile and quantitative assay for monitoring lysophagy in tissue culture cells.

Analysis of Lysophagic Flux in Cultured Cells using Lyso-Keima v2

2021 ◽  
Author(s):  
Vinay V. Eapen ◽  
Sharan Swarup ◽  
Melisa Hoyer ◽  
Harper JW
Keyword(s):  
Tissue Culture ◽  
Cultured Cells ◽  
Confocal Imaging ◽  
Signaling Proteins ◽  
Membrane Rupture ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Luminal Side ◽  
Galectin 3 ◽  
Autophagy Pathway

Lysophagy-the selective elimination of damaged lysosomes by the autophagy pathway-is a critical housekeeping mechanism in cells. This pathway surveils lysosomes and selectively demarcates terminally damaged lysosomes for elimination. Among the most upstream signaling proteins in this pathway are the glycan binding proteins-Galectins-which recognize N and O linked glycan chains on the luminal side of transmembrane lysosomal proteins. These glycosyl modifications are only accessible to galectin proteins upon extensive lysosomal membrane rupture and serve as a sensitive measure of lysosomal damage and eventual clearance by selective autophagy. Indeed, prior work has shown that immunofluorescence of Galectin-3 serves as a convenient proxy for lysophagic flux in tissue culture cells (Aits et al., 2015; Maejima et al., 2013). Here we describe a facile method for monitoring lysophagy using the acid sensitive fluorophore mKeima, affixed onto Galectin-3, which allows for the monitoring of lysophagic flux by Flow cytometry, Western blotting or Confocal imaging. This method, which we have termed Lyso-Keima, serves as a facile and quantitative assay for monitoring lysophagy in tissue culture cells.

scholarly journals Molecular Dissection of Cytokinesis by RNA Interference inDrosophila Cultured Cells

2002 ◽  
Vol 13 (7) ◽  
pp. 2448-2460 ◽  
Author(s):  
Maria Patrizia Somma ◽  
Barbara Fasulo ◽  
Giovanni Cenci ◽  
Enrico Cundari ◽  
Maurizio Gatti
Keyword(s):  
Cultured Cells ◽  
Contractile Ring ◽  
Membrane Behavior ◽  
Double Stranded Rna ◽  
Central Spindle ◽  
Culture Cells ◽  
Ring Assembly ◽  
Tissue Culture Cells ◽  
Genome Wide ◽  
Spindle Microtubules

We have used double-stranded RNA-mediated interference (RNAi) to study Drosophila cytokinesis. We show that double-stranded RNAs for anillin, acGAP,pavarotti, rho1, pebble,spaghetti squash, syntaxin1A, andtwinstar all disrupt cytokinesis in S2 tissue culture cells, causing gene-specific phenotypes. Our phenotypic analyses identify genes required for different aspects of cytokinesis, such as central spindle formation, actin accumulation at the cell equator, contractile ring assembly or disassembly, and membrane behavior. Moreover, the cytological phenotypes elicited by RNAi reveal simultaneous disruption of multiple aspects of cytokinesis. These phenotypes suggest interactions between central spindle microtubules, the actin-based contractile ring, and the plasma membrane, and lead us to propose that the central spindle and the contractile ring are interdependent structures. Finally, our results indicate that RNAi in S2 cells is a highly efficient method to detect cytokinetic genes, and predict that genome-wide studies using this method will permit identification of the majority of genes involved inDrosophila mitotic cytokinesis.

scholarly journals Functional analysis of desmoplakin domains: specification of the interaction with keratin versus vimentin intermediate filament networks.

1993 ◽  
Vol 123 (3) ◽  
pp. 691-705 ◽  
Author(s):  
T S Stappenbeck ◽  
E A Bornslaeger ◽  
C M Corcoran ◽  
H H Luu ◽  
M L Virata ◽  
...  
Keyword(s):  
Intermediate Filament ◽  
Cultured Cells ◽  
Sequence Similarity ◽  
Full Length ◽  
Carboxyl Terminus ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Carboxyl Terminal ◽  
Filament Networks ◽  
Vimentin Intermediate Filament

We previously demonstrated that truncated desmoplakin I (DP I) molecules containing the carboxyl terminus specifically coalign with and disrupt both keratin and vimentin intermediate filament (IF) networks when overexpressed in tissue culture cells (Stappenbeck, T. S., and K. J. Green. J. Cell Biol. 116:1197-1209). These experiments suggested that the DP carboxyl-terminal domain is involved either directly or indirectly in linking IF with the desmosome. Using a similar approach, we have now investigated the behavior of ectopically expressed full-length DP I in cultured cells. In addition, we have further dissected the functional sequences in the carboxyl terminus of DP I that facilitate the interaction with IF networks. Transient transfection of a clone encoding full-length DP I into COS-7 cells produced protein that appeared in some cells to associate with desmosomes and in others to coalign with and disrupt IF. Deletion of the carboxyl terminus from this clone resulted in protein that still appeared capable of associating with desmosomes but not interacting with IF networks. As the amino terminus appeared to be dispensable for IF interaction, we made finer deletions in the carboxyl terminus of DP based on blocks of sequence similarity with the related molecules bullous pemphigoid antigen and plectin. We found a sequence at the very carboxyl terminus of DP that was necessary for coalignment with and disruption of keratin IF but not vimentin IF. Furthermore, the coalignment of specific DP proteins along keratin IF but not vimentin IF was correlated with resistance to extraction by Triton. The striking uncoupling resulting from the deletion of specific DP sequences suggests that the carboxyl terminus of DP interacts differentially with keratin and vimentin IF networks.

scholarly journals Microinjected fluorescent polystyrene beads exhibit saltatory motion in tissue culture cells.

1984 ◽  
Vol 98 (6) ◽  
pp. 2126-2132 ◽  
Author(s):  
M C Beckerle
Keyword(s):  
Tissue Culture ◽  
Cultured Cells ◽  
Cytoplasmic Matrix ◽  
High Voltage Electron Microscopy ◽  
Polystyrene Beads ◽  
Voltage Electron ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
A Cell ◽  
Path Lengths

Microinjected 0.26-micron fluorescent, carboxylated microspheres were found to display classical saltatory motion in tissue culture cells. The movement of a given particle was characterized by a discontinuous velocity distribution and was unaffected by the activity of adjacent particles. The microspheres were translocated at velocities of up to 4.7 micron/s and sometimes exhibited path lengths greater than 20 micron for a single saltation . The number of beads injected into a cell could range from a few to over 500 with no effect on the cell's ability to transport them. Neither covalent cross-linking nor preincubation of the polystyrene beads with various proteins inhibited the saltatory motion of the injected particles. The motion of the injected beads in cultured cells was reversibly inhibited by the microtubule poison nocodazole, under conditions in which actin-rich, nitrobenzoxadiazol - phallacidin -staining structures remain intact. Whole-cell high voltage electron microscopy of microinjected cells that were known to be moving the fluorescent microspheres revealed that the beads were embedded in the cytoplasmic matrix and did not appear to be membrane bound. The enhanced detectability of the fluorescent particles over endogenous organelles and the ability to modify the surfaces of the beads before injection may enable more detailed studies on the mechanism of saltatory particle motion.

scholarly journals STUDIES ON THE INTRACELLULAR DIGESTIVE PROCESS IN MAMMALIAN TISSUE CULTURE CELLS

1965 ◽  
Vol 25 (2) ◽  
pp. 41-55 ◽  
Author(s):  
Gerald B. Gordon ◽  
Leonard R. Miller ◽  
Klaus G. Bensch
Keyword(s):  
Hydrolytic Enzymes ◽  
Multivesicular Bodies ◽  
Colloidal Iron ◽  
Gold Particles ◽  
Digestion Process ◽  
Culture Cells ◽  
Dense Bodies ◽  
Tissue Culture Cells ◽  
Golgi Region ◽  
Esterolytic Activity

DNA-protein coacervates containing colloidal gold particles were readily phagocytized by strain L fibroblasts. During the subsequent digestion process, the gold particles served as markers which permitted the demonstration of the evolution of digestive vacuoles to multivesicular bodies and finally to dense bodies. Acid phosphatase and esterolytic activity was present in these structures. The hydrolytic enzymes were apparently brought to the phagocytotic vacuoles in small vesicles originating in the Golgi region. These vesicles entered the vacuoles prior to the digestion of the coacervates and the appearance of positive cytochemical reactions. The cytoplasmic dense bodies frequently merged with the phagocytotic vacuoles. This was demonstrated by prelabeling the dense bodies with colloidal iron prior to phagocytosis of the coacervates. In addition, evidence is presented for the interrelationship of the phagocytotic and autophagic pathways.

Cytoplasmic Tubules in Cells Infected with Laryngotracheitis Virus

1969 ◽  
Vol 27 ◽  
pp. 376-377
Author(s):  
A. M. Watrach
Keyword(s):  
Tissue Culture ◽  
Small Proportion ◽  
Chicken Embryo ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Morphologic Characteristics ◽  
Infectious Laryngotracheitis ◽  
Laryngotracheitis Virus ◽  
In Cells

During a study of the development of infectious laryngotracheitis (LT) virus in tissue culture cells, unusual tubular formations were found in the cytoplasm of a small proportion of the affected cells. It is the purpose of this report to describe the morphologic characteristics of the tubules and to discuss their possible association with the development of virus.The source and maintenance of the strain of LT virus have been described. Prior to this study, the virus was passed several times in chicken embryo kidney (CEK) tissue culture cells.

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