Uptake and release of adenosine in isolated rat fat cells

Radioactively labelled adenosine and adenine were rapidly taken up by isolated rat fat cells, and incorporated into nucleotides, of which ATP dominated. The overall process had an apparent K_m of 1–5 μM. During incubation, especially in the presence of lipolytic agents, there was a reduction in labelled ATP with a compensatory increase in ADP, AMP, cAMP and nucleosides. The build-up of adenosine during incubation was inhibited by theophylline, which inhibits 5′-nucleotidase. Radioactivity released from perifused fat cells consisted mainly of nucleoside material, of which adenosine predominated. Lipolytic stimulation caused no significant increase in nucleoside outflow from perifused cells, whereas oxygenation was capable of reducing this outflow. It is concluded that adenosine is formed by fat cells as a consequence of ATP breakdown. Stimulation of lipolysis during activation of the sympathetic nerves leads to reversible ATP breakdown and adenosine release. Adenosine might therefore act as a modulator of lipolysis in vivo under these conditions, even though it does not serve as a feed back regulator in the proper sense.     

Uptake and release of adenosine in isolated rat fat cells.

Radioactively labelled adenosine and adenine were rapidly taken up by isolated rat fat cells, and incorporated into nucleotides, of which ATP dominated. The overall process had an apparent Km of 1--5 micrometers. During incubation, especially in the presence of lipolytic agents, there was a reduction in labelled ATP with a compensatory increase in ADP, AMP, cAMP and nucleosides. The build-up of adenosine during incubation was inhibited by theophylline, which inhibits 5'-nucleotidase. Radioactivity released from perifused fat cells consisted mainly of nucleoside material, of which adenosine predominated. Lipolytic stimulation caused no significant increase in nucleoside outflow from perifused cells, whereas oxygenation was capable of reducing this outflow. It is concluded that adenosine is formed by fat cells as a consequence of ATP breakdown. Stimulation of lipolysis during activation of the sympathetic nerves leads to reversible ATP breakdown and adenosine release. Adenosine might therefore act as a modulator of lipolysis in vivo under these conditions, even though it does not serve as a feed back regulator in the proper sense.     

Regional variation of white adipocyte lipolysis during the annual cycle of the alpine marmot.

During winter, hibernating animals rely on their lipid stores for survival. In vitro lipolytic activity of isolated adipocytes from gonadal and subcutaneous white adipose tissue (WAT) was studied in captive alpine marmots (Marmota marmota) at two different times of their yearly cycle. During the summer, when marmots were eating, adipocyte responsiveness and sensitivity to isoprenaline and noradrenaline were higher in gonadal than in subcutaneous WAT. During hibernation, when marmots were spontaneously fasting. both the response and sensitivity to catecholamines decreased in gonadal WAT to the level of subcutaneous WAT. A similar pattern of response was also observed when lipolysis was stimulated with glucagon but the lipolytic rate was three times lower than with catecholamines. Adenosine deaminase (ADA) had a marked stimulatory effect on lipolysis, especially during the 'feeding' period, suggesting that adenosine may be a potent lipolytic modulator in marmot adipocytes. It is concluded that in marmots, lipolysis could be differentially regulated between fat depots during the annual cycle possibly to optimize either the building-up or the use of fat reserves.     

Accumulation and inactivation of adenosine by fat cells from hypothyroid rats

It has been suggested that the deficient lipolytic response to catecholamines in hypothyroidism may be due to an increased sensitivity to adenosine and/or increased adenosine levels in this condition. We confirmed that the addition of adenosine deaminase enhanced the lipolytic response of hypothyroid fat cells, but the stimulation was at least as large in euthyroid cells. Adenosine analogs were more potent as antagonists of NA-induced lipolysis in hypothyroid than in euthyroid fat cells, but the difference could be explained by a decreased response to NA. Suspensions of hypothyroid cells accumulated more purine nucleosides (115 +/- 20) than did euthyroid cells (48 +/- 8 pmol/30 min/10(5) cells; p less than 0.01). This difference could not be explained by a lower rate of adenosine elimination, which occurred by three different pathways: uptake followed by phosphorylation, uptake followed by deamination and deamination by the serum albumin preparation. Under certain circumstances the latter pathway is of overwhelming importance. Fat cells from mature rats (460-480 g) behaved similarly as cells from young control rats. Thus, the changes induced by hypothyroidism was not due to a developmental change. The results are discussed in relation to earlier findings on the alterations in catecholamine responsiveness in hypothyroidism. It is concluded that an increased influence of adenosine could possibly explain some aspects of altered catecholamine responsiveness. If it does the mechanism is likely to involve an enhanced amount of adenosine rather than an increased sensitivity to adenosine.     

Cyclic AMP-dependent and independent inhibition of lipolysis by adenosine and decreased pH.

NA-stimulated lipolysis and cAMP formation in isolated rat fat cells is inhibited by acidosis. In the present report we have examined the quantitative relationship between lipolysis and cAMP formation at normal and reduced pH and the possible involvement of adenosine, an endogenous inhibitor of cAMP formation. Adenosine antagonized cAMP accumulation and to a considerably lower degree lipolysis, effects potentiated by acidosis. Theophylline, an antagonist of adenosine effects, stimulated lipolysis and cAMP-accumulation, and potentiated responses to NA. Adenosine deaminase (ADA) had theophylline-like effects. Acidosis inhibited lipolysis and cAMP accumulation induced by ADA and theophylline to a larger extent than those induced by NA. It is suggested that adenosine modulates fat cell cAMP production and may contribute to the antilipolytic effect of acidosis. There was a curvilinear relationship between cAMP elevation and glycerol production in fat cell suspensions, which was different at pH 7.4 and at pH 6.6. The amount of cAMP needed for half-maximal activation of lipolysis increased from 1.3 (pH 7.4) to 3.1 pMol X 10(-5) cells (pH 6.6). The maximal glycerol production was reduced from 1 300 to 900 nMol X 10(-5) cells. The antilipolytic effect of acidosis is apparently due partly to an inhibition of cAMP formation and partly to inhibition of subsequent step(s) in the activation sequence.     

Cyclic AMP-Dependent and Independent Inhibition of Lipolysis by Adenosine and Decreased pH

NA-stimulated lipolysis and cAMP formation in isolated rat fat cells is inhibited by acidosis. In the present report we have examined the quantitative relationship between lipolysis and cAMP formation at normal and reduced pH and the possible involvement of adenosine, an endogenous inhibitor of cAMP formation. Adenosine antagonized cAMP accumulation and to a considerably lower degree lipolysis, effects potentiated by acidosis. Theophylline, an antagonist of adenosine effects, stimulated lipolysis and cAMP-accumulation, and potentiated responses to NA. Adenosine deaminase (ADA) had theophylline-like effects. Acidosis inhibited lipolysis and cAMP accumulation induced by ADA and theophylline to a larger extent than those induced by NA. It is suggested that adenosine modulates fat cell cAMP production and may contribute to the antilipolytic effect of acidosis. There was a curvilinear relationship between cAMP elevation and glycerol production in fat cell suspensions, which was different at pH 7.4 and at pH 6.6. The amount of cAMP needed for half-maximal activation of lipolysis increased from 1.3 (pH 7.4) to 3.1 pMol × 10^-5 cells (pH 6.6). The maximal glycerol production was reduced from 1 300 to 900 nMol × 10^-5 cells. The antilipolytic effect of acidosis is apparently due partly to an inhibition of cAMP formation and partly to inhibition of subsequent step(s) in the activation sequence.     

Direct antilipolytic effect of acidosis in isolated rat adipocytes.

The possibility that acidosis inhibits lipolysis indirectly by causing ionic shifts or by favouring the accumulation of an inhibitor has been tested in isolated fat cells. Lipolysis induced by 3 muM noradrenaline (NA) was inhibited by 40-60% and that induced by 1 mM theophylline (THEO) by about 75% when the pH was reduced to 6.6. Lipolysis induced by NA + THEO was inhibited by 20-30%. Changing the concentration of Ca++ or Mg++ did not alter the degree of inhibition. Reducing the K+-ion concentration enhanced the inhibitory effect of low pH on lipolysis induced by NA or NA + THEO, whereas cyclic AMP accumulation was uninfluenced. Omitting glucose from the incubation medium caused a slight enhancement of pH-induced inhibition of lipolysis (from 60 to 70%, p less than 0.01). Reducing the concentration of albumin, which binds inhibitory substances such as FFA, reduced lipolysis more at normal than at reduced pH. At high FFA/albumin ratios (5 or above) lipolysis was similar at normal and reduced pH. The antilipolytic effect of decreased pH was equally pronounced in perifused fat cells, where inhibitory substances are not allowed to accumulate. Our results suggest that the antilipolytic effect of acidosis is mainly a direct effect of the increase in H+ ion concentration. The inhibitory effect of acidosis on various responses to beta-adrenoceptor stimulation may be caused by a decreased formation of cyclic AMP in turn caused directly by the decrease in pH.     

Direct Antilipolytic Effect of Acidosis in Isolated Rat Adipocytes

The possibility that acidosis inhibits lipolysis indirectly by causing ionic shifts or by favouring the accumulation of an inhibitor has been tested in isolated fat cells. Lipolysis induced by 3 μM noradrenaline (NA) was inhibited by 40–60% and that induced by 1 mM theophylline (THEO) by about 75% when the pH was reduced to 6.6. Lipolysis induced by NA+THEO was inhibited by 20–30%. Changing the concentration of Ca⁺⁺or Mg⁺⁺did not alter the degree of inhibition. Reducing the K⁺-ion concentration enhanced the inhibitory effect of low pH on lipolysis induced by NA or NA + THEO, whereas cyclic AMP accumulation was uninfluenced. Omitting glucose from the incubation medium caused a slight enhancement of pH-induced inhibition of lipolysis (from 60 to 70%, p<0.01). Reducing the concentration of albumin, which binds inhibitory substances such as FFA, reduced lipolysis more at normal than at reduced pH. At high FFA/albumin ratios (5 or above) lipolysis was similar at normal and reduced pH. The antilipolytic effect of decreased pH was equally pronounced in perifused fat cells, where inhibitory substances are not allowed to accumulate. Our results suggest that the antilipolytic effect of acidosis is mainly a direct effect of the increase in H⁺ion concentration. The inhibitory effect of acidosis on various responses to β-adrenoceptor stimulation may be caused by a decreased formation of cyclic AMP in turn caused directly by the decrease in pH.     

On the hormonal regulation of lipolysis in isolated reindeer adipocytes

The effects of insulin and bovine growth hormone (bGH) on lipolysis (basal and/or adrenaline-stimulated) were investigated in isolated adipocytes from the Norwegian reindeer. Addition of 100 nmol l-1 adrenaline to the cells increased lipolysis 20-fold above basal lipolytic rate. Insulin (0.I and I nmol l-I) depressed lipolysis significantly at submaximal adrenaline stimulation but had no effect when lipolysis was stimulated maximally. Bovine GH (25 and 100 ng ml-I) inhibited basal lipolysis during the first 2 h of incubation, whereas refractoriness to this action was induced during the third and fourth hour of incubation. These effects were not influenced by inclusion of a small amount of glucocorticoid (dexamethasone, 25 ng ml-I). Addition of 0.I and I nmol l-I insulin together with bGH (100 ng ml-I) prevented the initial antilipolytic effect of the latter and resulted in the expression of normal basal lipolytic rates. Preincubation with bGH (100 ng ml-I) for 2 h did not alter the lipolytic response of the adipocytes to adrenaline. These findings lend support to the conclusion that insulin exerts a major control of lipolysis in reindeer adipose tissue. The significance of the antilipolytic effect of bGH is difficult to evaluate since this effect was transient and was abolished during the third and fourth hour of incubation.     

The antilipolytic effect of endogenous and exogenous adenosine in canine adipose tissue in situ

The effects of adenosine, 2-Cl-adenosine, two adenosine uptake inhibitors (dipyridamole and dilazep) and the adenosine deaminase (ADA) inhibitor erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) were studied on basal and stimulated lipolysis in subcutaneous adipose tissue. The basal lipolysis was unaffected by all agents. Lipolysis induced by nerve stimulation (4 Hz, 5 min) was dose-dependently antagonized (up to 100%) by close i.a. infusions of adenosine (1–40 μM in blood); if the nerve induced vasoconstriction was prevented by α-adrenoceptor-blockade. 2-Cl-adenosine was a more potent antilipolytic agent than adenosine. EHNA (3–10 μM in blood) did not inhibit stimulated lipolysis in vivo possibly because of the low ADA activity in fat cells. Dipyridamole (0.5-1.5 μM in blood) in combination with EHNA increased the venous plasma concentration of adenosine from 0.3±0.05 to 0.7±0.1 μM and enhanced the tissue concentration close to 3-fold. Lipolysis induced by nerve stimulation (4 Hz) was reduced by about 40% by dipyridamole + EHNA and that induced by close i.a. noradrenaline injection (20 nmol) by approximately 60%. It is concluded that adenosine is an antagonist of stimulated lipolysis in subcutaneous adipose tissue in situ in concentrations that are reached during prolonged sympathetic nerve stimulation.     

Adipocyte dysfunction in rats with streptozotocin–nicotinamide‐induced diabetes

Administration of streptozotocin (STZ) and nicotinamide (NA) to adult rats allows for the induction of mild diabetes. However, this experimental model has not been fully characterized. This study was undertaken to determine the metabolic and secretory activity of adipose tissue in rats with STZ‐NA‐induced diabetes. Experiments were performed using epididymal adipocytes isolated from control and mildly diabetic rats. Lipogenesis, glucose transport as well as glucose and alanine oxidation, lipolysis, anti‐lipolysis, cAMP levels and adipokine secretion were compared in cells isolated from the control and diabetic rats. Lipogenesis, glucose transport and oxidation were diminished in the adipocytes of diabetic rats compared with the fat cells of control animals. However, alanine oxidation appeared to be similar in the cells of non‐diabetic and diabetic animals. Lipolytic response to low epinephrine concentrations was slightly increased in the adipocytes of diabetic rats; however, at higher concentrations of the hormone, lipolysis was similar in both groups of cells. The epinephrine‐induced rise in cAMP levels was higher in the adipocytes of STZ‐NA‐induced diabetic rats, even in the presence of insulin. Lipolysis stimulated by dibutyryl‐cAMP did not significantly differ, whereas anti‐lipolytic effects of insulin were mildly decreased in the cells of diabetic rats. Secretion of adiponectin and leptin was substantially diminished in the adipocytes of diabetic rats compared with the cells of control animals. Our studies demonstrated that the balance between lipogenesis and lipolysis in the adipose tissue of rats with mild diabetes induced by STZ and NA is slightly shifted towards reduced lipid accumulation. Simultaneously, adiponectin and leptin secretion is significantly impaired.     

Uptake of adenosine by dispersed chich embryonic cardiac cells.

Adenosine is involved in the regulation of coronary blood flow, but its mechanism of action is not clear. The present investigation is an attempt to understand the mechanism(s) of uptake of adenosine in dispersed chick embryonic cardiac cells and its relationship to the adenosine hypothesis. Adenosine is readily taken up by these cardiac cells, and a small fraction is incorporated into adenine nucleotides, whereas a major fraction is deaminated to inosine. The mechanism of uptake is different in 12- to 15-day-old chick embryos compared to 16- to 22-day-old embryos. The younger embryo heart cells show the incorporation of adenosine into adenine mononucleotides of the incubation medium as well as all the adenine nucleotides of the cells, whereas the older embryo heart cells show incorporation of adenosine only into the adenine nucleotides of the cells. The isolated cells used in the present study do not leak any significant amounts of adenosine kinase and/or nucleotides, and free adenosine was not found in the cells, even with extracellular concentrations as high as 1 mM. The absence of free adenosine in isolated dispersed cells reflects the activities of adenosine kinase and adenosine deaminase and is compatible with the adenosine hypothesis for the regulation of coronary blood flow.     

Adenosine mediated desensitization of cAMP signaling enhances T‐cell responses

Adenosine has long been regarded as a crucial anti‐inflammatory agent that protects the host from excessive damage. It has been reported to play an important role in suppressing immune activation, particularly that of T cells. However, it is a general observation that induction of T‐cell activation is an efficient event despite the high adenosine levels that are often present in the affected host due to injury or stress. We report here that prior to antigenic stimulation via TCR/CD3, exposure of T cells to adenosine desensitizes adenosine receptors, so as to create a window of time where the T cells are insensitive to this ubiquitous suppressor. T cells from mice that were pre‐exposed to this manipulation showed stronger responses to antigenic stimulation; therefore, the P1 adenosine receptor desensitization demonstrated an adjuvant‐like effect. Our results suggest that adenosine receptor desensitization may be a mechanism for T cells to escape the general suppression during early points of T‐cell activation and may emerge as a potential alternative for vaccine adjuvants.     

Lipolytic response of adipocytes to epinephrine in sedentary and exercise-trained subjects: sex-related differences

Summary Adipose tissue lipolytic activity is increased in endurance-trained subjects, but little is known about the mechanisms of this increase. To understand more fully the mechanisms involved and to discover whether sex-related differences exist, biopsies of fat were performed in the periumbilical region of 20 sedentary subjects (10 women (W) and 10 men (M)) and 20 trained subjects (10 W, 10 M); the in vitro response to epinephrine of the collagenase-isolated fat cells was studied. Glycerol release, chosen as an adipocyte lipolysis indicator, was measured by bioluminescence. Dose-response curves with epinephrine (α₂ and β agonist), with isoproterenol (β agonist) and epinephrine + propranolol and adenosine deaminase, were studied. Epinephrine-induced lipolysis was enhanced in trained subjects and this was due to an increased efficiency of the β-adrenergic pathway. However, differences were found between the two sexes. In trained men, the lipolysis increase resulted from the enhancement of the β-adrenergic pathway efficiency without any significant decrease in the α₂-adrenergic pathway efficiency. In trained women, the lipolysis increase was not only due to the enhancement of the β-adrenergic pathway efficiency (which was greater than in trained men), but also to a significant decrease in the α₂-adrenergic pathway efficiency. Despite the decrease, the α₂-adrenergic pathway remained more efficient in trained women than in trained men, as was the case in sedentary subjects. It is concluded that endurance training led to better lipid mobilization and that this effect seemed greater in women than in men.     

Adenosine potentiates the effects of LH and isoproterenol on luteal cells but not on isolated intact corpora lutea

Adenosine potentiated the stimulatory effect of luteinizing hormone (LH) in a dose-dependent way on the production of cyclic AMP (cAMP) in isolated cells from heavily luteinized rat ovaries and from individual rat corpora lutea of various ages (2- and 5-day-old). Such an effect has earlier been reported only for cells from heavily luteinized ovaries (Behrman et al. 1983). A similar potentiating effect of adenosine was now seen on catecholamine-stimulated cAMP production in isolated cells from 2-day-old corpora lutea. Adenosine did not, however, potentiate LH- or catecholamine-stimulated cAMP production in isolated intact corpora lutea.     

Ascorbic acid oral treatment modifies lipolytic response and behavioural activity but not glucocorticoid metabolism in cafeteria diet‐fed rats

Aim: To analyse the effects of vitamin C (VC), a potent dietary antioxidant, oral supplementation on body weight gain, behavioural activity, lipolytic response and glucocorticoid metabolism in the early stages of diet‐induced overweight in rats. Methods: Food intake, locomotive activity and faecal corticosterone were assessed during the 14 day trial period. After 2 weeks, the animals were sacrificed and the body composition, biochemical markers and lipolytic response from isolated adipocytes from retroperitoneal white adipose tissue were examined. Results: The intake of a high‐fat diet by rats induced a significant increase in body weight, adiposity and insulin resistance markers as well as a decrease in faecal corticosterone levels compared with standard diet‐fed rats. Interestingly, the animals fed on the cafeteria diet showed a significant increase in the isoproterenol‐induced lipolytic response in isolated adipocytes. Furthermore, this cafeteria‐fed group showed a reduced locomotive behaviour than the control rats. On the other hand, oral VC supplementation in animals receiving the high‐fat diet restored the cafeteria diet effect in some of the analysed variables such as final body weight and plasma insulin to control group levels. Remarkably, increases in locomotive behaviour and a significant decrease in the lipolytic response induced by isoproterenol on isolated adipocytes from animals treated with VC were observed. Conclusion: This work demonstrates that an oral ascorbic acid supplementation has direct effects on behavioural activity and on adipocyte lipolysis in early obesity stages in rats, which could indicate a protective short‐term role of this vitamin against adiposity induced by chronic high‐fat diet consumption.     

Enhanced activity of hormone sensitive lipase (HSL) in mesenteric but not epididymal fat correlates with higher production of epinephrine in mesenteric adipocytes in rat model of cachectic rheumatoid arthritis

Cachectic rheumatoid arthritis, the less frequent form of the disease, is associated with loss of fat mass and often more severe course of the disease. Its experimental model represents rat adjuvant arthritis (AA) characterized by edema, lack of appetite, sharp body weight and fat loss. As individual fat depots display functional differences, here we studied lipolytic activity and sensitivity to lipolytic stimuli of nodeless epididymal fat (eWAT) and perinodal mesenteric fat (mWAT) depots at the peak of AA. We also examined changes in catecholamine and cytokine levels involved in lipolysis in plasma and/or isolated adipocytes from both WATs to identify the contribution of local, adipocyte-based processes and/or systemic events to adiposity loss in cachectic rheumatoid arthritis. AA was induced to male Lewis rats by complete Freund’s adjuvant. Groups of ad libitum-fed and pair-fed controls were used to distinguish the effects of food restriction from inflammation-induced cachexia. Adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL) and its phosphorylated form (pHSL) were analyzed by western blot. CRP and catecholamine levels in plasma or adipocyte lysates were determined using ELISA kits. Cytokine-induced neutrophil chemoattractant-1 (CINC-1/CXCL1), monocyte chemoattractant protein-1 (MCP-1/CCL2), IL-1β, IL-6, IL-10 and leptin in adipocyte lysate were analyzed by quantitative protein microarray. Plasma glycerol and FFA were measured spectrophotometrically. AA rats developed severe cachexia, with lower adiposity in mWAT compared to normal and pair-fed controls, whereas in eWAT the adiposity was similarly reduced in AA and pair-fed groups. ATGL levels in both WATs were not affected by AA or pair feeding. AA upregulated levels of HSL, pHSL and pHSL/HSL ratio in mWAT, whereas none of these parameters has changed in eWAT of AA rats or in either WATs of pair-fed rats. In AA rats plasma glycerol was elevated, whereas FFA concentration was reduced. Plasma norepinephrine and epinephrine were increased in AA compared with both groups of controls. In eWAT adipocytes, AA but not pair feeding, upregulated norepinephrine levels. In mWAT adipocytes, AA rats showed higher epinephrine levels than pair-fed controls. Leptin levels in both WATs were depleted in AA animals in accordance with body weight loss. None of the measured cytokines in eWAT and mWAT was enhanced. Our results demonstrate augmented lipolytic activity in mWAT and not eWAT during cachectic arthritis. The adipocyte-derived cytokines do not seem to contribute to activated lipolysis. We first demonstrated enhanced presence of norepinephrine in perinodal adipocytes that may contribute to the regulation of local lipolytic activity by auto/paracrine fashion and thus provide independent fuel supply to activated lymph nodes.     

Adenosine and ATP effects on isolated guinea pig gallbladder

The effects of adenosine, ATP and several derivatives of adenosine were measured in isolated strips of guinea pig gallbladder. Adenosine caused relaxations which were antagonized by theophylline and potentiated by an inhibitor of adenosine uptake, 6-(1-hydroxy-5-nitrobenzylthio)-guanosine (HNBTG). Among several adenosine derivatives, 2-chloroadenosine and 5′-N-ethylcarboxymidoadenosine were similarly effective while 1-N⁶-phenylisopropyladenosine was only a weak relaxant. None of the derivatives caused maximal relaxations at 100 μM, and thus absolute potencies could not be determined. ATP caused predominantly contractile effects, with relaxations sometimes being evident at high concentrations. Indomethacin abolished contractile effects of ATP, suggesting prostaglandin involvement, and only relaxations were evident in its presence. Adenosine deaminase abolished the effects of adenosine and partly reduced the relaxant effects of ATP in the presence of indomethacin. In view of the low potency of adenosine and ATP, physiological roles for these compounds in gallbladder motility are not readily evident.     

Selective inhibition of cholera toxin- and catecholamine-stimulated lipolysis by blocking agents.

Vibrio cholerae enterotoxin stimulates lipolysis in rat epididymal fat cell suspensions. Like hormones this toxin increases adenylate cyclase activity, raising levels of cyclic adenosine 3',5'-monophosphate (cAMP), which activates a cellular lipase. Using specific blocking agents, we studied the responses to the adrenergic lipolytic hormones epinephrine, norepinephrine, and isoproterenol, and to cholera toxin. All stimulators were used at 100 x threshold dose. Propranolol (34 muM), a beta blocking agent, inhibited epinephrine stimulation (P less than 0.001) but not that of toxin (P greater than 0.2). Choleragenoid (25 mug/ml), a natural toxoid of cholera toxin, blocked stimulation by toxin (P less than 0.001) but not that of the adrenergic agents (P greater than 0.2). A beta blocker, practolol (3 mM), inhibited stimulation by the catecholamines tested (P less than 0.005) but not that of toxin (P greater than 0.05). Higher concentrations of propranolol (340 muM) and the alpha blocking agents phenoxybenzamine (3 mM) and phentolamine (1.6 mM) inhibited all agonists (P less than 0.001). The response to theophylline was inhibited by all blockers (P less than 0.05) except propranolol at the lower concentration (34 muM). A combined beta and alpha blockade using propranolol and epinephrine together did not inhibit toxin-mediated lipolysis. It appears that stimulation by cholera toxin is independent of beta adrenergic receptors. A major inhibition of theophylline-mediated lipolysis by alpha blocking drugs indicated a nonspecific effect of these agents at the concentrations used. The uninhibited response to toxin in the presence of propranolol and epinephrine suggests a lack of relationship of the toxin receptor to either alpha or beta receptors.     

Effect of adenosine, adenosine analogues and drugs inhibiting adenosine inactivation on lipolysis in rat fat cells

It has been suggested that adenosine may be a physiologically important modulator of lipolysis. In the present study it was found that adenosine inhibited lipolysis stimulated by low (0.03 micrometer) concentrations of noradrenaline (NA). Lipolysis stimulated by higher concentrations (0.3 and 3 micrometer) of NA was inhibited to a minor degree or not at all. Theophylline (1 micromete)-induced lipolysis was inhibited by adenosine (IC50 approximately 10 micrometer). Inhibition of theophylline-induced lipolysis was tested for several analogues of adenosine. Some N6-substituted adenosine analogues and 2-Cl-adenosine were more potent inhibitors. Adenine-nucleotides (ATP, ADP, AMP) were about equipotent with adenosine. Several adenosine analogues, including its breakdown products were considerably less potent or ineffective. None of the analogues tested inhibited the action of adenosine. Dipyridamol, dilazep and papaverine, which inhibit the uptake of adenosine into cells, caused only a slight enhancement of the antilipolytic effect of adenosine. None of the analogues inhibited the effect of adenosine. It is concluded that adenosine can inhibit lipolysis due to low, "physiological" concentrations of noradrenaline and of low concentration of theophylline via an action on a receptor structure on the cell surface which exhibits structural specificity.