Phalloidin fluorescence and confocal microscopy reveal the musculature development of clam Ruditapes philippinarum.

Abstract

Knowledge of early development in bivalves is of great importance to understand the function of animal organ systems and the evolution of phenotypic diversity. Manila clam (Ruditapes philippinarum) is an economically important bivalve living in marine intertidal zones. To determine the pattern of muscle development in the clams, we investigate the characteristics of musculature development utilizing phalloidin staining and confocal microscopy. Myofilaments first appear at the early trochophore stage, and gradually become orderly arranged during the transition from trochophore to veliger. For veliger, larval muscle system is mainly composed of dorsal velum retractors, medio-dorsal velum retractors, ventral velum retractors, ventral larval retractors and anterior and posterior adductor muscles. After metamorphosis, the muscle system of late veliger has been reconstructed, showing the irreversible shrink of velum retractor muscles, the rapid growth of wedge-shaped foot and mantle margins. One of the most striking changes in settled spats is the development of sophisticated architecture of foot musculature, which consists of transverse pedal muscles, anterior foot retractor and posterior foot retractor. These findings will not only provide the basis to understand the dynamic pattern of myogenesis in the burrowing bivalves, but also provide valuable information for comparative analysis of muscle development among bivalves.