MICROMERES ARE REQUIRED FOR NORMAL VEGETAL PLATE SPECIFICATION IN SEA-URCHIN EMBRYOS

Vegetal plate specification was assessed in S, purpuratus embryos afte r micromere deletions at the 4th, 5th and 6th cleavages, by assaying e xpression of the early vegetal plate marker Endo16, using whole-mount in situ hybridization, After 4th cleavage micromere deletions, the emb ryos typically displayed weak Endo16 expression in relatively few cell s of the lineages that normally constitute the vegetal plate, while af ter 5th and 6th cleavage micromere deletions the embryos exhibited str ong Endo16 expression in larger fractions of cells belonging to those lineages, When all four micromeres were deleted, the embryos were seve rely delayed in initiating gastrulation and sometimes failed to comple te gastrulation, However, if only one micromere was allowed to remain in situ throughout development, the embryos exhibited strong Endo16 ex pression and gastrulation occurred normally, on schedule with controls . Additional measurements showed that these microsurgical manipulation s do not alter cleavage rates or generally disrupt embryo organization , These results constitute direct evidence that the micromeres provide signals required by the macromere lineages for initiation of vegetal plate specification, The specification of the vegetal plate is complet ed in a normal manner only if micromere signaling is allowed to contin ue at least to the 6th cleavage stage.