Abstract
The mechanisms underlying sea urchin gastrulation and spiculogenesis have been sought for decades. We have identified two growth factor signaling pathways that are involved in these developmental events. Antibodies against mammalian platelet-derived growth factor (PDGF) receptor-β inhibited gastrulation and spiculogenesis, and antibodies against human epidermal growth factor (EGF) receptor disrupted gastrulation and spicule placement in Lytechinus pictus and L. variegatus embryos. Our studies suggested that the antibodies affect development by inhibiting rather than activating the signaling pathways. Polyclonal and monoclonal antibodies against the mammalian receptors recognized specifically Lytechinus proteins of the expected size of 170-180 x 103 M(r). Growth factor binding assays indicated that there are approximately 1.25 x 104 platelet-derived growth factor-like receptors per cell at the mesenchyme blastula stage of L. pictus, and human platelet-derived growth factor bound with an apparent affinity of K(D) = 4.4 nM to dissociated cells at the mesenchyme blastula stage. Immunolabelling experiments showed that at the gastrula stage, the Lytechinus platelet- derived growth factor-like receptors are located on the primary mesenchyme cells, the gut, and most prominently on the secondary mesenchyme cells and the stomodeum. The epidermal growth factor-like receptors stained less intensely on the gut and primary and secondary mesenchyme cells. Both receptors are expressed on the ciliary band and the gut of the pluteus larva but only the PDGF-like receptor is expressed on the primary mesenchyme cells. Pulse studies showed that the embryos are sensitive to the platelet-derived growth factor receptor-β and epidermal growth factor receptor antibodies from the blastula to sometime between the mesenchyme blastula and midgastrula stages. We show that antibodies enter the blastocoel as late as the gastrula stage. Our results suggest that platelet-derived growth factor-like and epidermal growth factor-like signaling pathways are involved in the early differentiation and morphogenesis of the sea urchin gut and spicules.