Lipid mobilization is increased during repeated bouts of exercise, but the underlying mechanisms involved in this process have not yet been delineated. The relative involvement of catecholamine-and ANP-dependent pathways in the control of lipid mobilization during repeated bouts of exercise was thus investigated in subcutaneous adipose tissue (SCAT) by microdialysis. The study was performed in healthy males. Subjects performed two 45-min exercise bouts (E1 and E2) at 50% of their maximal oxygen uptake separated by a 60-min rest period. Extracellular glycerol concentration (EGC), reflecting SCAT lipolysis, was measured in a control probe perfused with Ringer solution and in two other probes perfused with either Ringer plus phentolamine (␣1/2-AR antagonist) or Ringer plus both phentolamine and propranolol (-AR antagonist). Plasma epinephrine, plasma glycerol, and EGC were 1.7-, 1.6-, and 1.2-fold higher in E2 than in E1, respectively. Phentolamine potentiated exercise-induced EGC increase during E2 only. Propranolol reduced the lipolytic rate during both E1 and E2 compared with the probe with phentolamine. Plasma ANP concentration increased more during E2 than during E1 and was correlated with the increase in EGC in the probe containing phentolamine plus propranolol. The results suggest that ANP is involved in the control of lipolysis during exercise and that it contributes to stimulation of lipolysis during repeated bouts of exercise. microdialysis; insulin; epinephrine; guanosine 5Ј,5Ј-cyclic monophosphate UNTIL RECENTLY, the regulation of human adipose tissue lipolysis during exercise was mainly attributed to both the increase in catecholamine levels and the concomitant decrease in plasma insulin concentration. Catecholamines activate lipolysis through  1 -and  2 -adrenergic receptors (AR) and inhibit it through ␣ 2 -AR stimulation. The coordinated activation of both receptors modulates the intracellular cAMP concentration, which activates cAMP-dependent protein kinase, leading to the phosphorylation and activation of hormone-sensitive lipase (HSL) (2, 22). The recent discovery that natriuretic peptides (NP) are potent activators of lipolysis in human fat cells raises new questions about the control of lipid mobilization in humans (27,28). NP stimulate fat cell plasma membrane receptors (NPR-A subtype) bearing an intrinsic guanylyl cyclase activity and elevate intracellular levels of cGMP, which activate a cGMP-dependent protein kinase (PKG). PKG-dependent phosphorylation of perilipin and HSL stimulate lipolysis (27,28). NP are potent lipolytic agents when they act on isolated fat cells from subcutaneous adipose tissue (SCAT), specifically in primates (29). Intravenous infusion of pharmacological doses of hANP (human atrial natriuretic peptide) in humans promotes a strong lipid-mobilizing effect independently of reflex activation of the sympathetic nervous system (SNS) (12). A similar response occurred when ANP was perfused through a microdialysis probe inserted in SCAT (27). Moreover, within a physiological...