The swim bladder of Glaucosoma hebracium is specialized by having two side membranes extending dorsally from the side to the vertebrae. This characteristic supports the morphology of the Glaucosomatidae. There is a pair of reddish muscles connecting the skull and the swim bladder of the Glaucosoma hebracium. This type of muscle is likely to involve with sound production. However, vocalization has not been reported in the Glaucosomatids. As direct recordings for sound production in live specimens of this species could not easily be carried out, function of these swim bladder muscles was assessed by means of biochemical assay. Activities of Citrate synthase (CS) and L-lactate dehydrogenase (LDH), known to involve with energy supply during muscle contraction were assessed. Two-dimensional gels for the white muscle, tail region red muscle and swim bladder muscles were compared to reveal difference of protein expression. Results show that CS activity of the swim bladder muscle was higher than the white muscle. But LDH activities in these two types of muscles were not significant different. As high activities in CS reflecting aerobic nature of the tissue, the present dataset suggests that the swim bladder muscles should involve with sound production. Proteomics analyzes identified 14 proteins from the swim bladder muscle, one of them are related to energy production (ATP synthase beta chain). Two-dimensional gels show some different spots from these three type muscles. These results help us to find some different proteins in these three types of muscles of this fish. In conclusions base on these evidence in morphology, histology, enzyme activity assay and two-dimensional gels, the swim bladder muscles should involve with sound production in G. hebracium.