Summary
In the epididymal adipose tissue of lean (+/+) C57 BL/6J mice deprived of calcium by ethy-leneglycol-bis-(β-aminoethylether) N,N′-tetraacetic acid (EGTA) treatment, corticotrophin-(1-24)-tetracosapeptide (ACTH)-stimulated lipolysis was abolished and peak levels of adenosine 3′,5′-cyclic monophosphate (cyclic AMP1) were reduced by 80%. Calcium omission was less effective on isoproterenol-stimulated lipolysis and was ineffective on theophylline- or dibutyryl cyclic AMP-stimulated lipolysis. All EGTA effects were reversible. Lipolytic agents stimulated the net uptake of45Ca. This effect was inhibited by 2,4-dinitrophenol. The45Ca accumulated in response to lipolytic agents was largely La3+-nondisplaceable and was released with a half-life of 13 minutes under EGTA treatment. These results suggest that the mechanism promoting45Ca uptake in response to lipolytic agents was beyond cyclic AMP production in the adipose tissue of lean mice, but independent of free fatty acid release. In turn, intracellular as well as extracellular calcium facilitated maximal rates of lipolysis.
In the adipose tissue of obese-hyperglycaemic (ob/ob) mice, the response to the lipolytic agents of glycerol release and cyclic AMP levels was reduced by 65% and45Ca uptake was not stimulated. A 1–2 hour preincubation of adipose tissue increased markedly the effect upon lipolysis and45Ca uptake, but not that of peak cyclic AMP levels of isoproterenol. It is concluded that the adipose tissue of ob/ob mice displayed a reversible impairment of45Ca uptake in response to lipolytic agents.
Article PDF
Similar content being viewed by others
Abbreviations
- cyclic AMP:
-
adenosine 3′,5′-cyclic mono-phosphate
- ACTH:
-
corticotrophin-(1-24)-tetracosapeptide
- 2,4-DNP:
-
2,4-dinitrophenol
- HEPES:
-
N-2-hydroxyethyl-piperazine-N′-2-ethanesulfonic acid
- EGTA:
-
ethyleneglycol-bis-(β-aminoethylether)N,N′-tetraacetic acid
- ob/ob:
-
obesehyperglycaemic
References
Beifrage, P., Jergil, B., Strålfors, P., Tornqvist, H.: Hormone-sensitive lipase of rat adipose tissue: identification and some properties of the enzyme protein. FEBS Lett.75, 259–264 (1977)
Schwabe, U., Ebert, R.: Different effects of lipolytic hormones and phosphodiesterase inhibitors on cyclic 3′,5′-AMP levels in isolated fat cells. Naunyn Schmiedebergs Arch. Pharmacol.274, 287–298 (1972)
Dehaye, J.P., Winand, J., Christophe, J.: Lipolysis and cyclic AMP levels in epididymal adipose tissue of obese-hypergly-caemic mice. Diabetologia13, 553–561 (1977)
Fain, J. N.: Biochemical aspects of drug and hormone action on adipose tissue. Pharmacol. Rev.25, 67–118 (1973)
Siddle, K., Hales, C. N.: The action of local anaesthetics on lipolysis and on adenosine 3′,5′-cyclic monophosphate content in isolated rat fat cells. Biochem. J.142, 345–351 (1974)
Schimmel, R. J.: Calcium antagonists and lipolysis in isolated rat epididymal adipocytes: effects of tetracaine, manganese, cobaltous and lanthanum ions and D600. Horm. Metab. Res.10, 128–134 (1978)
Schimmel, R. J.: The influence of extracellular calcium ion on hormone activated lipolysis. Biochim. Biophys. Acta326, 272–278 (1973)
Mosinger, B., Vaughan, M.: Effects of electrolytes on epinephrine stimulated lipolysis in adipose tissue in vitro. Biochim. Biophys. Acta144, 556–568 (1967)
Akgün, S., Rudman, D.: Relationships between mobilization of free fatty acids from adipose tissue, and the concentration of calcium in the extracellular fluid and in the tissue. Endocrinology84, 926–930 (1969)
Alm, B., Efendić, S., Löw, H.: Effect of lipolytic and antilipolytic agents on the uptake of47calcium into rat adipose tissue in vitro. Horm. Metab. Res.2, 142–146 (1970)
Efendic, S., Alm, B., Löw, H.: Effects of Ca++ on lipolysis in human omental adipose tissue in vitro. Horm. Metab. Res.2, 287–291 (1970)
Dehaye, J. P., Winand, J., Christophe, J.: Adenylate cyclase activity in the epididymal adipose tissue from obese-hyperglycaemic mice. Diabetologia15, 45–51 (1978)
Shepherd, R.E., Malbon, C.C., Smith, C.J., Fain, J.N.: Lipolysis and adenosine 3′∶5′-monophosphate metabolism in isolated white fat cells from genetically obese-hyperglycemic mice (ob/ob). J. Biol. Chem.252, 7243–7248 (1977)
Dehaye, J. P., Winand, J., Christophe, J.: Effect of lipolytic agents on calcium uptake in the epididymal adipose tissue of obese-hyperglycemic mice and their lean littermates. Arch. Int. Physiol. Biochim.85, 396–398 (1977)
Wieland, O.: Eine enzymatische Methode zur Bestimmung von Glycerol. Biochem. Z.329, 313–319 (1957)
Gilman, A. G.: A protein binding assay for adenosine 3′,5′-cyclic monophosphate. Proc. Natl. Acad. Sci. USA67, 305–312 (1970)
Bihler, I., Jeanrenaud, B.: ATP content of isolated fat cells. Effects of insulin, ouabain and lipolytic agents. Biochim. Biophys. Acta202, 496–506 (1970)
Van Breemen, C., Farinas, B.R., Casteels, R., Berba, P., Wuytack, F., Deth, R.: Factors controlling cytoplasmic Ca2+ concentration. Proc. R. Soc. Lond. [Biol.]265, 57–71 (1973)
Hellman, B.: Calcium and pancreatic β-cell function. 3. Validity of the La3+-wash technique for discriminating between superficial and intracellular45Ca. Biochim. Biophys. Acta540, 534–542 (1978)
Katocs, A. S., Jr., Largis, E. R., Allen, D. O.: Role of Ca2+ in adrenocorticotropic hormone-stimulated lipolysis in the perifused fat cell system. J. Biol. Chem.249, 2000–2004 (1974)
Khoo, J. C., Steinberg, D., Thompson, B., Magee, S. E.: Hormonal regulation of adipocyte enzymes. The effects of epinephrine and insulin on the control of lipase, phosphorylase kinase, phosphorylase and glycogen synthetase. J. Biol. Chem.248, 3823–3830 (1973)
Fain, J. N.: Inhibition of adenosine cyclic 3′,5′-monophosphate accumulation in fat cells by adenosine N6-(phenyl-iso-propyl) adenosine and related compounds. Mol. Pharmacol.9, 595–604 (1973)
Angel, A., Desai, K. S., Halperin, M. L.: Reduction in adipocyte ATP by lipolytic agents: relation to intracellular free fatty acid accumulation. J. Lipid Res.12, 203–213 (1971)
Siddle, K., Hales, C. N.: Calcium and lipolysis in adipose tissue: effects of ethanedioxybis-(ethylamine)tetra-acetate (EGTA) and ionophore A23187. Biochem. Soc. Trans.5, 959–962 (1977)
Martin, B. R., Clausen, T., Gliemann, J.: Relationships between the exchange of calcium and phosphate in isolated fatcells. Biochem. J.152, 121–129 (1975)
Winand, J., Christophe, J.: Métabolisme in vitro du tissu adipeux. IV. Relations entre les espaces aqueux du tissu adipeux épididymaire du rat et l'albumine sérique. Bull. Soc. Chim. Biol.49, 299–315 (1967)
Crofford, O. B., Renold, A. E.: Glucose uptake by incubated rat epididymal adipose tissue. Rate-limiting steps and site of insulin action. J. Biol. Chem.240, 14–21 (1965)
Gliemann, J., Österlind, K., Vinten, J., Gammeltoft, S.: A procedure for measurement of distribution spaces in isolated fat cells. Biochim. Biophys. Acta286, 1–9 (1972)
Bruns, D.E., McDonald, J.M., Jarett, L.: Energy-dependent calcium transport in endoplasmic reticulum of adipocytes. J. Biol. Chem.251, 7191–7197 (1976)
Hales, C. N., Luzio, J. P., Chandler, J. A., Herman, L.: Localization of calcium in the smooth endoplasmic reticulum of rat isolated fat cells. J. Cell Sci.15, 1–15 (1974)
Severson, D. L., Denton, R. M., Bridges, B. J., Randle, P. J.: Exchangeable and total calcium pools in mitochondria of rat epididymal fat-pads and isolated fat-cells. Biochem. J.154, 209–223 (1976)
Clausen, T.: Electrolytes and the hormonal control of organic metabolism in adipocytes. In: Adipose tissue. Regulation and metabolic functions. Jeanrenaud, B., Hepp, D. (Eds.), pp. 66–70. New York, London: Academic Press 1970
Hope-Gill, Kissebah, A., Tulloch, B., Clarke, P., Vydelingum, N., Fraser, T. R.: The effects of insulin on adipocyte calcium flux and the interaction with the effects of dibutyryl cyclic AMP and adrenaline. Horm. Metab. Res.7, 195–196 (1975)
Rasmussen, H.: Cell communication, calcium ion, and cyclic adenosine monophosphate. Science170, 404–412 (1970)
Hems, D. A., Ma, G. Y.: Resistance to hepatic action of vasopressin in genetically obese (ob/ob) mice. Biochem. J.160, 23–28 (1976)
Keppens, S., Vandenheede, J. R., De Wulf, H.: On the role of calcium as second messenger in liver for the hormonally induced activation of glycogen phosphorylase. Biochim. Biophys. Acta496, 448–457 (1977)
York, D. A., Bray, G. A., Yukimura, Y.: An enzymatic defect in the obese (ob/ob) mouse: Loss of thyroid-induced sodium- and potassium-dependent adenosinetriphosphatase. Proc. Natl. Acad. Sci. USA75, 477–481 (1978)
Clarke, P. V., Kissebah, A. H., Hope-Gill, H., Vydelingum, N., Tulloch, B., Fraser, T. R.: The role of calcium in insulin action. IV. Mechanism of insulin resistance in adipose tissue of obese (ob/ob) mice and old Wistar rats. Eur. J. Clin. Invest.5, 351–358 (1975)
Winand, J., Christophe, J.: Effect of preincubation on the metabolism of [U-14C] glucose and [3H] H2O by the epididymal adipose tissue of obese-hyperglycemic (ob/ob) Bar Harbor mice. Arch. Int. Physiol. Biochim.84, 207–208 (1976)
Freychet, P., Laudat, M.H., Laudat, P., Rosselin, G., Kahn, C. R., Gorden, P., Roth, J.: Impairment of insulin binding to the fat cell plasma membrane in the obese-hyperglycemic mouse. FEBS Lett.25, 339–342 (1972)
Yen, T. T., Steinmetz, J. A.: Lipolysis of genetically obese and/or hyperglycemic mice with reference to insulin response of adipose tissue. Horm. Metab. Res.4, 331–337 (1972)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Dehaye, J.P., Winand, J., Poloczek, P. et al. Relationship between lipolysis and calcium in epididymal adipose tissue of obese-hyperglycaemic mice. Diabetologia 16, 399–408 (1979). https://doi.org/10.1007/BF01223161
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01223161