The present study aimed to analyze adiposity heterogeneity and the role of liver X receptor (LXRα) and peroxisome proliferator-activated receptors (PPARs) as targets of tumour necrosis factor-α (TNFα) in gilthead sea bream (Sparus aurata L.). The screening of 20 fish at the beginning of the warm season identified two major groups with fat and lean phenotypes. Fat fish showed increased liver and mesenteric fat depots. This increased adiposity was concurrent in the adipose tissue to enhanced expression of lipoprotein lipase (LPL) whereas mRNA levels of the hormone-sensitive lipase (HSL) remained almost unchanged. The resulting LPL/HSL ratio was thereby highest in fat fish, which suggests that this group of fish has not reached its peak fat storage capacity. This is not surprising given the increased expression of PPARγ in the absence of a counter-regulatory raise of TNFα. However, this lipolytic cytokine exerted dual effects in primary adipocyte cultures that differ within and between lean and fat fish. One set of fat fish did not respond to TNFα treatment whereas a second set exhibited a lipolytic response (increased glycerol release) that was apparently mediated by the downregulated expression of PPARβ. In lean fish, TNFα exerted a strong and non-transcriptionally mediated lipolytic action. Alternatively, TNFα would inhibit lipid deposition via the downregulated expression of adipogenic nuclear factors (PPARγ and LXRα). TNFα targets are therefore different in fish with lean and fat phenotypes, which is indicative of the complex network involved in the regulation of fish lipid metabolism.