Abstract
Anin vitro cultured rat perirenal preadipocyte (PA) was established as a model system to investigate the role of the intracellular pH (pHi) and of the Na+ /H+ exchanger during PA proliferation and differentiation. pH sensitive probe, 2′,7′-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein(BCECF), was employed to measure the pHi of PA and to determine the Na+/H+ exchange activity. The results showed that there was Na+/H+ exchange activity in the plasma membrane of PA, FCS stimulated DNA synthesis measured by3H-TdR incorporation, and the activation of Na+ /H+ exchanger resulted in pHi increase (nearly 0.2 pH unit) within 2 min. Ethyl-isopropyl-amiloride (EIPA), a specific Na+/H+ exchange inhibitor, inhibited Na+/H+ exchange activity and DNA synthesis. In the absence of serum insulin did not stimulate DNA synthesis but did induce PA differentiation characterized by the appearance of adiposome in the cell and the enhancement of glyeerol-3-phosphate dehydrogenase (G3PDHase) activity. Meantime, insulin was also found to stimulate Na+/H+ exchange activity and pHi increase. EIPA inhibited Na+/H+ exchanger activation induced by insulin and also partially inhibited the enhancement of G3PDHase activity. These results demonstrated that the activation of Na+ /H+ exchange and the resulting pHi increase are the early events related to both proliferation and differentiation of PA.
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Abbreviations
- BCECF:
-
2′,7′-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein
- FCS:
-
fetal calf serum
- EIPA:
-
ethyl-isopropyl-amiloride
- PKC:
-
protein kinase C
- PMA:
-
phorbol-12-myristate-13-acetate
- BSA:
-
bovine serum albumin
- G3PDHase:
-
glycerol-3-phosphate dehydrogenase
References
Sardet, C., Counillon, L., Franchi, A. et al., Growth factors induce phosphorylation of the Na+ /H+ antiporter, a glycoprotein of 110 KD,Science, 1990, 247: 723.
Grinstein, S., Rotin, D., Mason, M. J., Na+/H+ exchange and growth factor-induced cytosolic pH changes, role in cellular proliferation,Biochim. Biophys. Acta, 1989, 988: 73.
Chen, Q., Benson, R. S. P., Whetton, A. D. et al., Role of acid/base homeostasis in the suppression of apoptosis in haemopoietic cells by v-Abl protein tyrosine kinase,J. of Cell Sci., 1997, 110: 379.
Smyth, M. J., Sparks, R. L., Wharton, W., Proadipocyte cell lines: models of cellular proliferation and differentiation,J. Cell Sci., 1993, 106: 1.
Shillabeer, G., Forden, J. M., Lau, D. C. W., Induction of preadipocyte differentiation by ature fat cells in the rat,J. Clin. Invest., 1989, 84: 381.
Li, Z.H., Lu, Z.D., Kirkland, J. L. et al., Preadipocyte stimulating factor in rat serum: evidence for a discrete 63 KDa protein that promote cell differentiation of rat preadipocytes in primary cultures,J. Cell. Physiol., 1989, 141: 543.
Read, S. M., Northcote, D. H., Minimization of variation in the response to different proteins of the coomassie blue G dye-binding assay for protein,Anal. Biochem., 1981, 116: 53.
Daniel, T. O., Ives, H. E., Cyclosporin A inhibits kinase C-independent activation of the Na+ /H+ exchanger by PDGF and vanadate,Biochem. Biophy. Res. Commu., 1987, 145(1): 111.
L’Allemain, G., Franchi, A., Cragoe, E. et al., Blockade of the Na+/H+ antiport abolishes growth factor-induced DNA synthesis in fibroblasts,J. Biol. Chem., 1984, 259(7): 4313.
Grinstein, S., Rothstein, A., Mechanisms of regulation of Na+/H+ exchanger,J. Membr. Biol., 1986, 90: 1.
Arsenis, G., Spencer, B. A., Regulation of Na+/H+ exchange in rat adipocytes; effects of insulin,Endocrinology, 1995, 136: 1920.
Yamaguchi, Y. F., Kathuria, S., Characterization of receptor tyrosine-specific protein kinases by the use of inhibitors. Staurosporine is a 100 times more potent inhibitor of insulin receptor than IGF-I receptor,Biochem. Biophy. Res. Commu., 1988, 157(3): 955.
Arsenis, G., Spencer, B. A., Ewert, M. S., Activation of Na+ /H+ exchanger by cellular pH and extracellular Na+ in rat adipocytes; inhibition by isoproterenol,Endocrinology, 1995, 136: 3128.
Ladoux, A., Cragoe, E. J., Geny, B. et al., Differentiation of human promyelocytic HL 60 ells by retinoic acid is accompanied by an increase in the intracellular pH,J. Biol. Chem., 1987, 262(2): 811.
Dyck, J. R. B., Fliegel, L., Specific activation of the Na+/H+ exchanger gene during neuronal differentiation of embryonal carcinoma cells,J. Biol. Chem., 1995, 270: 10420.
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Project supported partially by the Foundation of State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Zoology, the Chinese Academy of Sciences and the Foundation of Institute of Zoology, the Chinese Academy of Sciences.
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Chen, Q., Li, X., Lu, Z. et al. Activation of Na+ /H+ exchange on rat preadipocyte plasma membrane and its role in cell proliferation and differentiation. Sci. China Ser. C.-Life Sci. 42, 240–248 (1999). https://doi.org/10.1007/BF03183599
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DOI: https://doi.org/10.1007/BF03183599