scholarly journals Incorporation and Rate of Loss of S35-Methionine by Adult Rat Trachea in Organ Cultures and in Living Animals

1960 ◽  
Vol 7 (2) ◽  
pp. 283-288 ◽  
Author(s):  
T. Timothy Crocker ◽  
S. R. Pelc

Full-thickness pieces of adult rat trachea were supported on rayon on the surface of clotted medium in watch glasses. Differentiated epithelium was reduced in height during 25 days of cultivation because basal cells and some columnar cells migrated to cover exposed parts of the explants and because some differentiated cells died and were shed. S35-methionine was (a) placed on explants in vitro and (b) injected intraperitoneally in living rats. Cultured tissues and tissues of living rats were examined by autoradiography at 4 and 24 hours and 4, 7, and 11 days after labeling. Although migratory undifferentiated epithelial cells appeared in cultured trachea, all living epithelial cells in vitro incorporated and subsequently lost S35-methionine to the same extent as did epithelium of intact rats. The biologic half-life of methionine in rat tracheal epithelium in vivo and in vitro was about 5 days.

2007 ◽  
Vol 176 (2) ◽  
pp. 147-154 ◽  
Author(s):  
Terry Lechler ◽  
Elaine Fuchs

Despite their importance in cell shape and polarity generation, the organization of microtubules in differentiated cells and tissues remains relatively unexplored in mammals. We generated transgenic mice in which the epidermis expresses a fluorescently labeled microtubule-binding protein and show that in epidermis and in cultured keratinocytes, microtubules stereotypically reorganize as they differentiate. In basal cells, microtubules form a cytoplasmic network emanating from an apical centrosome. In suprabasal cells, microtubules concentrate at cell–cell junctions. The centrosome retains its ability to nucleate microtubules in differentiated cells, but no longer anchors them. During epidermal differentiation, ninein, which is a centrosomal protein required for microtubule anchoring (Dammermann, A., and A. Merdes. 2002. J. Cell Biol. 159:255–266; Delgehyr, N., J. Sillibourne, and M. Bornens. 2005. J. Cell Sci. 118:1565–1575; Mogensen, M.M., A. Malik, M. Piel, V. Bouckson-Castaing, and M. Bornens. 2000. J. Cell Sci. 113:3013–3023), is lost from the centrosome and is recruited to desmosomes by desmoplakin (DP). Loss of DP prevents accumulation of cortical microtubules in vivo and in vitro. Our work uncovers a differentiation-specific rearrangement of the microtubule cytoskeleton in epidermis, and defines an essential role for DP in the process.


2000 ◽  
Vol 48 (4) ◽  
pp. 523-533 ◽  
Author(s):  
Emmanuel Mongodin ◽  
Odile Bajolet ◽  
Jocelyne Hinnrasky ◽  
Edith Puchelle ◽  
Sophie de Bentzmann

Staphylococcus aureus is a common human pathogen involved in non-bronchial diseases and in genetic and acquired bronchial diseases. In this study, we applied an immunolabeling approach for in vivo and in vitro detection of S. aureus, based on the affinity of staphylococcal protein A (SpA) for the Fc region of immunoglobulins, especially IgG. Most strains of S. aureus, including clinical strains, can be detected with this labeling technique. The approach can be used for detection and localization with transmission electron microscopy or light-fluorescence microscopy of S. aureus in infected tissues such as human bronchial tissue from cystic fibrosis (CF) patients. The methodology can also be applied to cell culture models with the aim of characterizing bacterial adherence to epithelial cells in backscattered electron imaging with scanning electron microscopy. Application to the study of S. aureus adherence to airway epithelium showed that the bacteria did not adhere in vivo to intact airway epithelium. In contrast, bacteria adhered to the basolateral plasma membrane of columnar cells, to basal cells, to the basement membrane and were identified beneath the lamina propria when the epithelium was injured and remodeled, or in vitro when the epithelial cells were dedifferentiated.


1993 ◽  
Vol 21 (2) ◽  
pp. 191-195 ◽  
Author(s):  
Knut-Jan Andersen ◽  
Erik Ilsø Christensen ◽  
Hogne Vik

The tissue culture of multicellular spheroids from the renal epithelial cell line LLC-PK1 (proximal tubule) is described. This represents a biological system of intermediate complexity between renal tissue in vivo and simple monolayer cultures. The multicellular structures, which show many similarities to kidney tubules in vivo, including a vectorial water transport, should prove useful for studying the potential nephrotoxicity of drugs and chemicals in vitro. In addition, the propagation of renal epithelial cells as multicellular spheroids in serum-free culture may provide information on the release of specific biological parameters, which may be suppressed or masked in serum-supplemented media.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Katharina Ernst ◽  
Ann-Katrin Mittler ◽  
Veronika Winkelmann ◽  
Carolin Kling ◽  
Nina Eberhardt ◽  
...  

AbstractWhooping cough is caused by Bordetella pertussis that releases pertussis toxin (PT) which comprises enzyme A-subunit PTS1 and binding/transport B-subunit. After receptor-mediated endocytosis, PT reaches the endoplasmic reticulum from where unfolded PTS1 is transported to the cytosol. PTS1 ADP-ribosylates G-protein α-subunits resulting in increased cAMP signaling. Here, a role of target cell chaperones Hsp90, Hsp70, cyclophilins and FK506-binding proteins for cytosolic PTS1-uptake is demonstrated. PTS1 specifically and directly interacts with chaperones in vitro and in cells. Specific pharmacological chaperone inhibition protects CHO-K1, human primary airway basal cells and a fully differentiated airway epithelium from PT-intoxication by reducing intracellular PTS1-amounts without affecting cell binding or enzyme activity. PT is internalized by human airway epithelium secretory but not ciliated cells and leads to increase of apical surface liquid. Cyclophilin-inhibitors reduced leukocytosis in infant mouse model of pertussis, indicating their promising potential for developing novel therapeutic strategies against whooping cough.


1985 ◽  
Vol 40 (4) ◽  
pp. 297-302 ◽  
Author(s):  
David R. Mann ◽  
Diane Evans ◽  
Festus Edoimioya ◽  
Freja Kamel ◽  
George M. Butterstein

2007 ◽  
Vol 67 (9) ◽  
pp. 4364-4372 ◽  
Author(s):  
Mathew Casimiro ◽  
Olga Rodriguez ◽  
Llana Pootrakul ◽  
Maral Aventian ◽  
Nadia Lushina ◽  
...  

1984 ◽  
Vol 30 (3) ◽  
pp. 381-388 ◽  
Author(s):  
B. R. Merrell ◽  
R. I. Walker ◽  
S. W. Joseph

The initial interaction and adherence of Vibrio parahemolyticus to epithelial tissue culture cells, human buccal epithelial cells, and the ileal mucosa of mice were studied. Using scanning electron microscopy, adherent bacteria were observed only on degenerating human embryonic intestinal, HeLa, and buccal cells; healthy normal cells were devoid of bacteria. Sheared V. parahaemolyticus, i.e., lacking flagella, did not adhere to either normal or degenerating tissue cells. Neither ultraviolet-inactivated organisms nor cell-free culture supernate affected the epithelial cells. Similar findings were observed on the mucosa of the ileum in mice inoculated with V. parahaemolyticus. It appears that V. parahaemolyticus possesses a cytotoxic factor which alters epithelial cells. This factor appears to be closely associated with viable organisms and may be a functional element in the adherence process of flagellated V. parahaemolyticus to mammalian epithelial cells.


2012 ◽  
Vol 130 (6) ◽  
pp. 1375-1383 ◽  
Author(s):  
Jin-Ah Park ◽  
Asma S. Sharif ◽  
Daniel J. Tschumperlin ◽  
Laurie Lau ◽  
Rachel Limbrey ◽  
...  

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