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.

scholarly journals FUNCTIONS OF COATED VESICLES DURING PROTEIN ABSORPTION IN THE RAT VAS DEFERENS

1967 ◽  
Vol 35 (2) ◽  
pp. 357-376 ◽  
Author(s):  
Daniel S. Friend ◽  
Marilyn G. Farquhar
Keyword(s):  
Electron Microscopy ◽  
Golgi Complex ◽  
Vas Deferens ◽  
Hydrolytic Enzymes ◽  
Rat Vas Deferens ◽  
Coated Vesicles ◽  
Multivesicular Bodies ◽  
Dense Bodies ◽  
Apical Cytoplasm ◽  
Golgi Region

The role of coated vesicles during the absorption of horseradish peroxidase was investigated in the epithelium of the rat vas deferens by electron microscopy and cytochemistry. Peroxidase was introduced into the vas lumen in vivo. Tissue was excised at selected intervals, fixed in formaldehyde-glutaraldehyde, sectioned without freezing, incubated in Karnovsky's medium, postfixed in OsO4, and processed for electron microscopy. Some controls and peroxidase-perfused specimens were incubated with TPP,1 GP, and CMP. Attention was focused on the Golgi complex, apical multivesicular bodies, and two populations of coated vesicles; large (> 1000 A) ones concentrated in the apical cytoplasm and small (<750 A) ones found primarily in the Golgi region. 10 min after peroxidase injection, the tracer is found adhering to the surface plasmalemma, concentrated in bristle-coated invaginations, and within large coated vesicles. After 20–45 min, it is present in large smooth vesicles, apical multivesicular bodies, and dense bodies. Peroxidase is not seen in small coated vesicles at any interval. Counts of small coated vesicles reveal that during peroxidase absorption they first increase in number in the Golgi region and later, in the apical cytoplasm. In both control and peroxidase-perfused specimens incubated with TPP, reaction product is seen in several Golgi cisternae and in small coated vesicles in the Golgi region. With GP, reaction product is seen in one to two Golgi cisternae, multivesicular bodies, dense bodies, and small coated vesicles present in the Golgi region or near multivesicular bodies. The results demonstrate that (a) this epithelium functions in the absorption of protein from the duct lumen, (b) large coated vesicles serve as heterophagosomes to transport absorbed protein to lysosomes, and (c) some small coated vesicles serve as primary lysosomes to transport hydrolytic enzymes from the Golgi complex to multivesicular bodies.

scholarly journals DISTRIBUTION OF PHOSPHATASES IN THE GOLGI REGION AND ASSOCIATED STRUCTURES OF THE THORACIC GANGLIONIC NEURONS IN THE GRASSHOPPER, MELANOPLUS DIFFERENTIALIS

1968 ◽  
Vol 37 (1) ◽  
pp. 89-104 ◽  
Author(s):  
Nancy J. Lane
Keyword(s):  
Acid Phosphatase ◽  
Phosphatase Activity ◽  
Acid Phosphatase Activity ◽  
Spatial Association ◽  
Thiamine Pyrophosphate ◽  
Multivesicular Bodies ◽  
Dense Bodies ◽  
Developmental Relationship ◽  
Golgi Region ◽  
Melanoplus Differentialis

The neuronal perikarya of the grasshopper contain sudanophilic lipochondria which exhibit an affinity for vital dyes. These lipochondria are membrane-delimited and display acid phosphatase activity; hence they correspond to lysosomes. Unlike those of most vertebrates, these lysosomes also hydrolyze thiamine pyrophosphate and adenosine triphosphate. Like vertebrate lysosomal "dense bodies," they are electron-opaque and contain granular, vesicular, or lamellar material. Along with several types of smaller dense bodies, they are found in close spatial association with the Golgi apparatus. The Golgi complexes are frequently arranged in concentric configurations within which these dense bodies lie. Some of the smaller dense bodies often lie close to or in association with the periphery of dense multivesicular bodies. Further, bodies occur that display gradations in structure between these multivesicular bodies and the dense lysosomes. Acid phosphatase activity is present in the small as well as the larger dense bodies, in the multivesicular bodies, and in some of the Golgi saccules, associated vesicles, and fenestrated membranes; thiamine pyrophosphatase is found in both the dense bodies and parts of the Golgi complex. The close spatial association of these organelles, together with their enzymatic similarities, suggests the existence of a functional or developmental relationship between them.

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.

scholarly journals SECRETION AND ENDOCYTOSIS IN INSULIN-STIMULATED RAT ADRENAL MEDULLA CELLS

1973 ◽  
Vol 56 (2) ◽  
pp. 540-558 ◽  
Author(s):  
Susan J. Abrahams ◽  
Eric Holtzman
Keyword(s):  
Acid Phosphatase ◽  
Horseradish Peroxidase ◽  
Adrenal Medulla ◽  
Lipid Droplets ◽  
Secretory Granules ◽  
Multivesicular Bodies ◽  
Dense Bodies ◽  
Golgi Region

Insulin was used to deplete the adrenalin stores of rat adrenal medulla cells. Release of secretion was observed to occur by exocytosis. In addition, during the stages of massive release of secretory granules, the insulin-treated preparations showed greatly enhanced endocytic uptake of horseradish peroxidase. The tracer was taken up within vesicles, tubules, multivesicular bodies, and dense bodies. From acid phosphatase studies and from previous work it appears that many of the structures in which peroxidase accumulates are lysosomes or are destined to fuse with lysosomes. Subsequent to the period of intense exocytosis and endocytosis, there is a transient accumulation of lipid droplets in the adrenalin cells. The cells then regranulate, with new granules forming near the Golgi region. These results suggest that under the conditions used, much of the membrane that initially surrounds secretory granules is degraded after release of the granules.

Ultrastructure of Liver in Lactosyl Ceramidosis

1971 ◽  
Vol 29 ◽  
pp. 312-313
Author(s):  
Z. Hruban ◽  
J. R. Esterly ◽  
G. Dawson ◽  
A. O. Stein
Keyword(s):  
Connective Tissue ◽  
Liver Biopsy ◽  
Osmium Tetroxide ◽  
Close Contact ◽  
Multivesicular Bodies ◽  
Bile Canaliculi ◽  
Dense Bodies ◽  
Marginal Plates ◽  
Membranous Material ◽  
Perichromatin Granules

Samples of a surgical liver biopsy from a patient with lactosyl ceramidosis were fixed in paraformaldehyde and postfixed in osmium tetroxide. Hepatocytes (Figs. 1, 2) contained 0.4 to 2.1 μ inclusions (LCI) limited by a single membrane containing lucid matrix and short segments of curved, lamellated and circular membranous material (Fig. 3). Numerous LCI in large connective tissue cells were up to 11 μ in diameter (Fig. 2). Heterogeneous dense bodies (“lysosomes”) were few and irregularly distributed. Rough cisternae were dilated and contained smooth vesicles and surface invaginations. Close contact with mitochondria was rare. Stacks were small and rare. Vesicular rough reticulum and glycogen rosettes were abundant. Smooth vesicular reticulum was moderately abundant. Mitochondria were round with few cristae and rare matrical granules. Golgi complex was seen rarely (Fig. 1). Microbodies with marginal plates were usual. Multivesicular bodies were very rare. Neutral lipid was rare. Nucleoli were small and perichromatin granules were large. Small bile canaliculi had few microvilli (Fig. 1).

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.

Sulfhydryl bond formation is a prerequisite for proper cycling of centrosomes and chromosomes in mammalian tissue culture cells

1993 ◽  
Vol 51 ◽  
pp. 342-343
Author(s):  
Heide Schatten ◽  
Neidhard Paweletz ◽  
Ron Balczon
Keyword(s):  
Tissue Culture ◽  
Cell Cycle Progression ◽  
Group Formation ◽  
Sulfhydryl Group ◽  
Mammalian Tissue ◽  
Mitotic Chromosomes ◽  
Microtubule Network ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Transmission Electron

To study the role of sulfhydryl group formation during cell cycle progression, mammalian tissue culture cells (PTK2) were exposed to 100¼M 2-mercaptoethanol for 2 to 6 h during their exponential phase of growth. The effects of 2-mercaptoethanol on centrosomes, chromosomes, microtubules, membranes and intermediate filaments were analyzed by transmission electron microscopy (TEM) and by immunofluorescence microscopy (IFM) methods using a human autoimmune antibody directed against centrosomes (SPJ), and a mouse monoclonal antibody directed against tubulin (E7). Chromosomes were affected most by this treatment: premature chromosome condensation was detected in interphase nuclei, and the structure in mitotic chromosomes was altered compared to control cells. This would support previous findings in dividing sea urchin cells in which chromosomes are arrested at metaphase while the centrosome splitting cycle continues. It might also support findings that certairt-sulfhydryl-blocking agents block cyclin destruction. The organization of the microtubule network was scattered probably due to a looser organization of centrosomal material at the interphase centers and at the mitotic poles.

UDP-D-glucose 4-epimerase activity of the tissue culture of white poplars (Populus alba L. var. pyramidalis)

1982 ◽  
Vol 47 (5) ◽  
pp. 1530-1536 ◽  
Author(s):  
Ladislav Bilisics ◽  
Štefan Karácsonyi ◽  
Marta Kubačková
Keyword(s):  
Tissue Culture ◽  
Cell Walls ◽  
Enzyme Preparation ◽  
Uridine Diphosphate ◽  
Populus Alba ◽  
Activity Change ◽  
White Poplar ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Galactose Content

The presence of UDP-D-glucose 4-epimerase (EC 5.1.3.2) in the culture tissue of white poplar was evidenced. As found, the partially purified enzyme preparation contained UDP-D-glucose glucosyltransferase, UDP-D-galactose galactosyltransferase and non-specific enzymes able to cleave the uridine-diphosphate saccharides into the appropriate hexose monophosphates. The activity change of UDP-D-glucose 4-epimerase in tissue culture cells during the growth was in accord with changes in D-galactose content in cell walls and indicated the possibility to regulate the formation of polysaccharides containing D-galactose at the level of production of UDP-D-galactose in cells.

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