Hormone-sensitive lipase (HSL) is an important regulator of cellular lipid homeostasis and catalyzes the hydrolysis of stored triacylglycerol. In this study, the full-length cDNA sequence of HSL of Macrobrachium nipponense (M. nipponense) was identified and cloned for the first time from a hepatopancreas cDNA library by using reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends. The complete HSL sequence was 3575 bp and encoded a 785 amino acid peptide with the catalytic core (GXSXG) containing a serine residue. The phylogenetic tree revealed that the gene of HSL of M. nipponense is closely related with that of Penaeus vanmamei. The tissue distribution showed that the mRNA expression level of HSL in the hepatopancreas were significantly higher than that in other tissues (P < 0.05). Furthermore, the quantitative real-time PCR results showed that HSL expression in the hepatopancreas was upregulated by an increase in dietary lipids and is partially inhibited when the ratio of phospholipids was increased in the lipid mixture. These results demonstrate that HSL is a novel gene involved in lipid metabolism of M. nipponensis and that HSL expression varies depending on the type of lipid.