Summary
A method for evaluation of microtubule content in cultured cells has been developed. The method is based on scanning microfluorometric measurement of immunofluorescently labelled microtubules. The method has been applied to the comparison of microtubule content in epithelial XTH-2 cells grown in culture at various cell densities. The results have shown that the microtubule content in the cells is not dependent on their proliferative state rather than it depends on cellular contacts.
Similar content being viewed by others
References
Baker JB, Humphreys T (1971) Serum stimulated release of cell contact and the initiation of growth in contact-inhibited chick fibroblasts. Proc Natl Acad Sci USA 68:2161–2164
Bereiter-Hahn J, Kajstura J (1987) Scanning microfluorimetric measurement of TRITC-phalloidin labelled F-actin in cultured cells: dependence of F-actin content on density of normal and transformed cells. Histochemistry (submitted)
Bereiter-Hahn J, Fava P, Fuhrmann C, Kajstura J, Münnich A (1987) What happens to culture cells when they become confluent? Eur J Cell Biol 43 (Suppl) 17:6
Bernd A, Schröder HC, Zahn RK, Müller WEG (1982) Modulation of the nuclear-envelope nucleoside triphosphatase by poly (a)-rich mRNA and by microtubule protein. Eur J Biochem 129:43–49
Bockus BJ, Stiles CD (1984) Regulation of cytoskeletal architecture by platelet-derived growth factor, insulin and epidermal growth factor. Exp Cell Res 153:186–197
Chafouelas JG, Pardue RL, Brinkley BR, Dedman JR, Means AR (1981) Regulation of intracellular levels of calmodulin and tubulin in normal and transformed cells. Proc Natl Acad Sci USA 78:996–1000
Crossin KL, Carney DH (1981) Evidence that microtubule depolymerization early in the cell cycle is sufficient to initiate DNA synthesis. Cell 23:61–71
Herman B, Pledger WJ (1985) Platelet-derived growth factor-induced alterations in vinculin and actin distribution in BALB/c-3T3 cells. J Cell Biol 100:1031–1040
Hiller G, Weber K (1978) Radioimmunoassay for tubulin: a quantitative comparison of the tubulin content of different established tissue culture cells and tissues. Cell 14:795–804
Kajstura J, Bereiter-Hahn J (1987) Scanning microfluorimetric measurement of cell constituents: principles of the method and its application to the determination of NAD content and redox state of XTH-2 cells in culture. Histochemistry (in press)
Kajstura J, Korohoda W (1983) Significance of energy metabolism pathways for stimulation of DNA synthesis by cell migration and serum. Eur J Cell Biol 31:9–14
Kilmartin JV, Wright B, Milstein C (1982) Rat monoclonal antitubulin antibodies derived by using a new nonsecreting rat cell line. J Cell Biol 93:576–582
Puck TT (1977) Cyclic AMP, the microtubule-microfilament system, and cancer. Proc Natl Acad Sci USA 74:4491–4495
Rose GG (1954) A separable and multipurpose tissue culture chamber. Tex Rep Biol Med 12:1074–1083
Rozengurt E (1985) The mitogenic response of cultured 3T3 cells: integration of early signals and synergistic effects in a unified framework. In: Cohen P, Houslay MD (eds) Molecular mechanisms of transmembrane signalling. Elsevier, Amsterdam, pp 429–454
Schlage WK (1986) Determination of human factor VIII (AHG-associated protein)-antigen in endothelial cells from Xenopus laevis (XTH cells). Evaluation of a sensitive ELISA technique. Cell Tissue Res 245:445–451
Schlage W, Bereiter-Hahn J, Kuhn C (1981) Established Xenopus tadpole heart endothelium (XTH) cells exhibiting selected properties of primary cells. Eur J Cell Biol 24:342
Wehland J, Schröder HC, Weber K (1984) Amino-acid sequence requirements in the epitope recognized by the α-tubulin-specific rat monoclonal antibody YL1/2. EMBO J 3:1295–1300
Wiche G, Lundbad VJ, Cole RD (1977) Competence of soluble cell extracts as microtubule assembly systems. J Biol Chem 252:794–796
Wolf K, Quimby MC (1962) Synthetic media. Amphibian culture medium. Excerpta Medica 16, Nr. 9. Excerpta Medica, Amsterdam
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kajstura, J., Bereiter-Hahn, J. Scanning microfluorometric measurement of immunofluorescently labelled microtubules in cultured cells. Histochemistry 88, 53–55 (1987). https://doi.org/10.1007/BF00490167
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00490167