Chicken embryo fibroblasts

Kelley, P.M. Schlesinger, M.J. (1978). The effect of amino acid analogues and heat shock gene expression in chicken embryo fibroblasts. Cell 15, 1277-1286. 

Bond, U. Schlesinger, M.J. (1985). Ubiquitin is a heat shock protein in chicken embryo fibroblasts. Molecular and Cellular Biology, 5, 949-56. 

Collier, N., and Schelsinger, M. J. (1986). The dynamic state of heat shock proteins in chicken embryo fibroblasts. J. Cell Biol. 103, 1495—1507. 

Weissmahr, R. N., Schupbach, J., and Boni, J. (1997) Reverse transcriptase activity in chicken embryo fibroblast culture supernatants is associated with particles containing EAV-0 (endogenous avian retrovirus) RNA. J. Virol. 71, 3005-3012. 

Liotti FS, Pelliccia C, Pezzetti F. 1988. Different response of chicken embryo fibroblasts and hepatocytes to the interference of certain antioxidants on the binding of [G- H] benzo[a]pyrene to DNA. Cancer Lett 41 235-242. 

Nl. Nakamura, K. D., Martinez, R., and Weber, M. J., Tyrosine phosphorylation of specific proteins after mitogen stimulation of chicken embryo fibroblasts. Mol. Cell. Biol. 3,380-390 (1983). 

Singer and colleagues (68, 69) have successfully visualized cytoskeletal mRNAs and their associate proteins using biotinylated cDNA probes followed by antibodies to biotin and collodial gold-conjugated antibodies. Their method used in situ hybridization followed by whole mount TEM of Triton-extracted chicken embryo fibroblasts. Cytoskeletal mRNAs were found in close proximity to actin protein and further from tubulin filaments. While the whole mount technology does limit the technique to extracted cells, applications to thin sections will allow greater resolution. 

Berg T, Cohen SB, Deshamais J et al (2002) Small-molecule antagonists of Myc/Max dimerization inhibit Myc-induced transformation of chicken embryo fibroblasts. Proc Natl Acad Sci USA 99 3830-3835... 

Bosca, L., Rousseau, G.G. Hue, L. (1985). Phorbol 12-myristate 13-acetate and inSulin increase the concentration of fructose-2,6-bisphosphate and stimulate glycolysis in chicken embryo fibroblasts. Proc. Natl Acad, Sci. USA, 82, 6440-4. 

Fukui, Y., Saltiel, AR. Hanafusa, R (1991). Phosphatidylinoatol-3 kinase is activated in v-src, v-yes, and v-fps transformed chicken embryo fibroblasts. Oncogene, 6, 407—11. 

Hughes, M., Sehgal, A., Hadman, M. Bos, T. (1992). Heterodimerization with c-Fos is not required for cell transformation of chicken embryo fibroblasts by Jun. Cell Growth Diff., 3, 889-97. 

Nermut, M.V., Burt, J.S., Hirst, E.MA. Larjava, R (1993). Distribution of avian integrin during the lifetime of chicken embryo fibroblasts in vitro study by immunofluorescence and immune electron microscopy. Micron, 24, 363-75. 

Iba H, Takeya T, Cross FR, Hanafusa T, Hanafusa H. Rous sarcoma vims variants that carry the cellular src gene instead of the viral src gene cannot transform chicken embryo fibroblasts. Proc Natl Acad Sci USA. 1984 81 4424-8. 

Erikson E, Stefanovic D, Blenis J, Erikson RL, Mailer JL. 1987. Antibodies to Xenopus egg S6 kinase II recognize S6 kinase from progesterone- and insulin-stimulated Xenopus oocytes and from proliferating chicken embryo fibroblasts. Mol Cell Biol 7(9) 3147-3155. 

Kinetic Studies of the ADP-Ribosylation of HSP-83 in Chicken Embryo Fibroblast Cells... 

Chicken embryo fibroblast cells (CEF) were prepared from the carcasses of 10-day-old embryos and maintained in complete growth medium as described [5]. The cells were used between the third and seventh subcultivation after initiation of the cultures. Cells were labeled in vivo with 125 juCi of [2,8,5 - H]adenosine per ml of culture media (50.5 Ci mmol 1 Ci = 3.7 x 10 Bq New England Nuclear) as described [5]. 

Chicken embryo fibroblasts were labeled with [ H]-adenosine for various lengths of time, the cells harvested, and the homogenates analyzed by one-dimensional SDS-PAGE and fluorography. Thereafter the amount of radioactivity incorporated into HSP-83 and various RNA species were directly quantitated as described in Materials and Methods. As illustrated in Fig. 1, the incorporation of tritium into HSP-83 is half-maximal saturated within 1 h of labeling of cells, while incorporation of [ H]-adenosine into tRNA and ribosomal 5S RNA continue to increase linearly for more than 24 h. The time it takes to reach saturation of labeling of HSP-83 resembles the time course for saturation of the ATP and NAD pools with radioactive precursor. This indicates that the actual modification of HSP-83 is not the rate limiting step and, thus, the turnover rate of the ADP-ribose moiety is very rapid. 

Chicken embryo fibroblasts (CEFs) are obtained from embryonic day-10 chicken embryos, and cultured as described (Albertinazzi et al, 1998). Transient expression of proteins is achieved by transfection of CEFs by the calcium phosphate technique using 10 fig of plasmid DNA for each 6 cm diameter plate. 18-24 h after transfection, cells are extracted with lysis buffer (1% Triton X-100,150 mM NaCl, 0.5 mg/ml phenyl methyl sulfonyl fluoride (PMSF), 20 mM Tris-Cl, pH 7.5). Cell lysates are clarified by differential centrifugation for 10 min at 18,000g in a refrigerated microcentrifuge, and utilized for analysis by sucrose velocity gradient centrifugation. 

Collier, N.C., Sheetz, M.P. Schlesinger, M.J. (1993) J. Cell Biochem. 52,297-397. Concomitant changes in mitochondria and intermediate filaments during heat shock and recovery of chicken embryo fibroblasts. 

---