Abstract
The present study was carried out to understand the effect of cortisol on calpain system in the C2C12 and 3T3-L1 adipocyte cells under co-culture system. Cells were co-cultured by using transwell inserts with a 0.4 μm porous membrane to separate C2C12 and 3T3-L1 preadipocyte cells. Each cell type was grown independently on the transwell plates. Following cell differentiation, inserts containing 3T3-L1 cells were transferred to C2C12 plates. Ten microgram per milliliter of cortisol was added to the medium. Following treatment for 3 days, the cells in the lower well were harvested for analysis. Calpains such as μ-calpain, m-calpain, and calpastatin were selected for the analysis. RT-PCR results indicated the significant increase in the mRNA expression of μ-calpain, m-calpain, and calpastatin. In addition, the confocal microscopical investigation indicated the cortisol treatment increases calpain expression in the C2C12 and 3T3-L1 cells. Taking all these together, cortisol treatment with co-culture system shows most reliable status of calpains expression in the cells, which is quite distinct from one-dimensional monocultured cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Koohmaraie, M., & Geesink, G. H. (2006). Meat Science, 74, 34–43.
Goll, D. E., Thompson, V. F., Li, H. Q., Wei, W., & Cong, J. Y. (2003). Physics Review, 83, 731–801.
Blomgren, K., Kawashima, S., Saido, T. C., Karlsson, J. O., Elmered, A., & Hagberg, H. (1995). Brain Research, 684, 143–149.
Squier, M. K. T., & Cohen, J. J. (1996). Cell Death and Differentiation, 3, 275–283.
Temm-Grove, C. J., Wert, D., Thompson, V. F., Allen, R. E., & Goll, D. E. (1999). Experimental Cell Research, 247, 293–303.
Dourdin, N., Bhatt, A. K., Dutt, P., Greer, P. A., Arthur, J. S., Elce, J. S., et al. (2001). Journal of Biological Chemistry, 276, 48382–48388.
Fox, J. E. (1999). Thrombosis and Haemostasis, 82, 385–391.
Glading, A., Lauffenburger, D. A., & Wells, A. (2002). Trends in Cell Biology, 12, 46–54.
Glading, A., Uberall, F., Keyse, S. M., Lauffenburger, D. A., & Wells, A. (2001). Journal of Biological Chemistry, 276, 23341–23348.
Melloni, E., Salamino, F., & Sparatore, B. (1992). Biochimie, 74, 217–223.
Saido, T. C., Nagao, S., Shiramine, M., Tsukaguchi, M., Yoshizawa, T., Sorimachi, H., et al. (1994). FEBS Letters, 346, 263–267.
Michetti, M., Salamino, F., Tedesco, I., Averna, M., Minafra, R., Melloni, E., et al. (1996). FEBS Letters, 392, 11–15.
Ilian, M. A., & Forsberg, N. E. (1992). Biochemistry Journal, 287, 163–171.
Dahlmann, B., Kuehn, L., Reinaue, H., Kay, J., & Stauber, W. T. (1989). Contributions to Nephrology, 73, 127–136.
Nelson, W. J., & Traub, P. (1983). Molecular and Cellular Biology, 3, 1146–1156.
Goll, D. E., Dayton, W. R., Singh, I., & Robson, R. M. (1991). Journal of Biological Chemistry, 266, 8501–8510.
Croall, D. E., & Demartino, D. N. (1991). Physiological Reviews, 71, 813–847.
Di Lisa, F., De Tullio, R., Salamino, F., Barbato, R., Melloni, E., Siliprandi, N., et al. (1995). Biochemistry Journal, 308(5), 7–61.
Goll, D. E., Kleese, W. C., & Szpacenko, A. (1989). In D. R. Campion, G. J. Hausman, & R. J. Martin (Eds.), Animal growth regulation (pp. 141–182). New York: Plenum Publishing Corporation.
Huang, J., & Forsberg, N. E. (1998). Proceedings of the National Academy of Sciences of the United States of America, 95, 12100–12105.
Fareed, M. U., Evenson, A. R., Wei, W., Menconi, M., Poylin, V., Petkova, V., et al. (2006). American Journal of Physiology - Regulatory, Integrative and Comparative Physiology, 290, R1589–R1597.
Cottin, P., Brustis, J. J., Poussard, S., Elamrani, N., Broncard, S., & Ducasting, A. (1994). Biochimica et Biophysica Acta, 1223, 170–178.
Patel, Y. M., & Lane, M. D. (1999). Proceedings of the National Academy of Sciences of the United States of America, 96, 1279–1284.
Ertbjerg, P., Henckel, P., Karlsson, A., Larsen, L. M., & M¢ller, A. J. (1999). Journal of Animal Science, 77, 2428–2436.
Sensky, P. L., Parr, T., Bardsley, R. G., & Buttery, P. J. (1996). Journal of Animal Science, 74, 380–387.
Ertbjerg, P., Moira, A. L., & Peter, P. P. (2000). Biochimie, 82, 197–201.
Sun, X., & Zemel, M. B. (2008). Journal of Nutritional Biochemistry, 19, 392–399.
Pfaffl, M. W. (2001). Nucleic Acids Research, 29, e45.
Goll, D. E., Thompson, V. F., Taylor, R. G., & Christiansen, J. A. (1992). Biochimie, 74, 225–237.
Goll, D. E., Otsuka, Y., Nagainis, P. A., Shannon, J. D., Sathe, S. K., & Muguruma, M. (1983). Journal of Food Biochemistry, 7, 137–177.
Higgins, J. A., Lasslett, Y. V., Bardsley, R. G., & Buttery, P. J. (1988). British Journal of Nutrition, 60, 645–652.
Koohmaraie, M., Shackelford, S. D., Muggli-Cockett, N. E., & Stone, R. T. (1991). Journal of Animal Science, 69, 4823–4835.
Mersmann, H. J. (1989). In D. R. Campion, G. J. Hausman, & R. J. Martin (Eds.), Animal growth regulation (pp. 337–357). New York: Plenum Publishing Corporation.
Iizuka, K., Kawaguchi, H., & Yasuda, H. (1991). Japanese Circulation Journal, 55, 1086–1093.
Dayton, W. R., Schollmeyer, J. V., Chan, A. C., & Allen, C. E. (1979). Biochimica et Biophysica Acta, 584, 216–230.
Arakawa, N., Takashima, M., Kurata, T., & Fujimaki, M. (2000). Agricultural and Biological Chemistry, 47, 1517–1522.
Parr, T., Sensky, P. L., Arnold, M. K., Bardsley, R. G., & Buttery, P. J. (2000). Archives of Biochemistry and Biophysics, 374, 299–305.
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Muthuraman, P., Ravikumar, S., Muthuviveganandavel, V. et al. Effect of Cortisol on Calpains in the C2C12 and 3T3-L1 Cells. Appl Biochem Biotechnol 172, 3153–3162 (2014). https://doi.org/10.1007/s12010-014-0753-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12010-014-0753-1