Parvalbumin characteristics in the sonic muscle of a freshwater ornamental grunting toadfish (Allenbatrachus grunniens)

Abstract

The grunting toadfish, Allenbatrachus grunniens, is an ornamental fish in freshwater aquariums, and it has the ability to produce sounds. The sonic muscle of the toadfish is the fastest vertebrate muscle ever measured, and the rates of Ca²⁺ transport and cross-bridge dissociation are also the fastest. Parvalbumins (PAs) are Ca²⁺-binding proteins that help in muscle relaxation in vertebrates. Several PA isoforms have been identified in variable ratios in different muscle types. Both male and female grunting toadfish have intrinsic sonic muscles attached to their swim bladders, but no significant difference in morphology between male and female sonic muscles has been observed. In this study, we used SDS-PAGE and western blotting to characterize the total PA expression and to identify the PAs from the sonic muscle and the white body muscle of A. grunniens. Although the total PA concentrations were similar in sonic and white muscles, there were differences in the isoform percentages. Two and four PA isoforms were identified from sonic muscle and white muscle, respectively. The estimated sizes of PA1, PA2, and PA3 in the sonic muscle of the grunting toadfish were 10, 10.5, and 10.5 kDa, respectively, and the isoelectric points of PA1, PA2, and PA3 in the grunting toadfish were 4.77, 4.58, and 4.42, respectively. In the sonic muscle, the primary PA isoform was PA1, which comprised more than 94 % of total PA, whereas PA2 comprised only 5 % of the total PA content. In contrast, in white muscle, the primary isoform was PA2, which comprised 58 % of the total PA. Both PA1 (with PA1a) and PA3 represented approximately 20 % of the total PA in white muscle. These results indicate that there is no positive correlation between a high PA content and the speed of muscle relaxation; however, PA1 might have the greatest effect on the relaxation of the grunting toadfish’s sonic muscle.