Adipose triglyceride lipase (ATGL) was recently identified as a triglyceride (TG)-specific lipase. In this study, we first obtained from large yellow croaker fish a 1,820-bp (GenBank ID: HQ916211) ATGL cDNA fragment with a 141-bp 5'UTR, a 1,485-bp open reading frame, and a 194-bp 3'UTR. The predicted fish ATGL had 494 amino acids (GenBank ID: ADY89608) and a calculated molecular weight of 55.1 kDa. ATGL was highly expressed in liver and, to a lesser degree, in heart, muscle, and abdominal fat. ATGL gene expression was high at 4.5 g and then decreased at 157.9 g and increased again at 474.2 g. The effects of lipid levels and lipid sources on ATGL expression in vivo were also investigated. A high-lipid diet decreased ATGL expression in fish significantly (P < 0.01). Fish in soybean oil group exhibited significantly lower ATGL expression than fish in the fish oil and beef tallow groups (P < 0.01). These data enhance our understanding of ATGL in fish lipid metabolism.