Lineage tracing of the bivalve shell field with special interest in the descendants of the 2d blastomere

By evolving bilaterally separated shell plates, bivalves acquired a unique body plan in which their soft tissues are completely protected by hard shell plates. In this unique body plan, mobility between the separated shell plates is provided by novel structures such as a ligament and adductor muscles. As a first step towards understanding how the bivalve body plan was established, we investigated the development of the separated shell plates and ligament. Over 100 years ago, it was hypothesized that the development of separated shell plates is tightly linked with the unique cell cleavage (division) pattern of bivalves during development, wherein each bilateral daughter cell of the 2d descendant 2d(1121) develops into one of the bilateral shell fields. In this study, we tested this hypothesis by tracing the cell lineages of the Japanese purple mussel Septifer virgatus. Although the shell fields were found to be exclusively derived from the bilateral descendant cells of 2d: 2d(11211) and 2d(11212), the descendants of these cells were not restricted to shell fields alone, nor were they confined to the left or right side of the shell field based on their lineage. Our study demonstrated that ligament cells are also derived from 2d(11211) and 2d(11212), indicating that the ligament cells emerged as a subpopulation of shell field cells. This also suggests that the establishment of the novel developmental system for the ligament cells was critical for the evolution of the unique body plan of bivalves.