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Study of Larval and Adult Skeletogenic Cells in Developing Sea Urchin Larvae

Tateyama Marine Laboratory, Marine and Coastal Research Center, Ochanomizu University, 11 Koyatsu, Tateyama, Chiba 294-0301, Japan

^* To whom correspondence should be addressed. E-mail: kiyomoto{at}cc.ocha.ac.jp

The larval skeleton of sea urchin embryos is formed by primary mesenchyme cells (PMCs). Thereafter, the larvae start feeding and additional arms develop. An adult rudiment that contains spines, tube feet, tests, and other parts of the adult body is formed in the eight-armed larva. The cellular mechanism of the later skeletogenesis and the lineage of the adult skeletogenic cells are not known. In this study, the morphogenesis of larval and adult skeletons during larval development of the sea urchin Hemicentrotus pulcherrimus was investigated by immunostaining cells with PMC-specific monoclonal antibodies, which are useful markers of skeletogenic cells. All spicules and the associated cells in the later larvae were stained with the antibodies. We could observe the initiation of skeletal morphogenesis at each developmental stage and visualize the cellular basis of skeleton formation in whole-mount embryos that possessed an intact morphology. There were some similarities between PMCs and the later skeletogenic cells. Both had a rounded shape with some filopodia, and the antigen expression started just before overt spicule formation. In the later-stage embryos, cells with filopodia and faint antigen expression were observed migrating in the blastocoel or aggregating in the presumptive location of new skeletogenesis.

Abbreviations: PBSC, phosphate-buffered saline saturated with calcite • PMC, primary mesenchyme cell • SMC, secondary mesenchyme cell • SW, seawater