In vitro developmental toxicity of concanavalin A in rat embryos: analysis of neural crest cell migration using monoclonal antibody HNK-1.
The developmental toxicity of concanavalin A (Con A) was evaluated in vitro using a rat whole embryo culture system, and the distributions of the neural crest cells were immunohistochemically investigated in embryos with monoclonal antibody HNK-1. In addition, binding sites of Con A in the embryos were observed according to the avidin-biotin peroxidase complex method with biotin-labeled anti-Con A. The rat embryos on embryonic day 8 were exposed to a final concentration of 12.5, 25, 50, or 100 micrograms/ml of Con A for a 72 hr culture period. Exposure to Con A concentration-dependently resulted in lower viability, decreases in yolk sac diameter, crown-rump length, and number of somites, and increases in the incidence of morphological abnormalities characterized by neural tube defects in the embryos. In the Con A-treated embryos, the distributions of the neural crest cells were restricted in the dorsal and cranial regions, and the migration into the interventricular chamber was delayed in the cardiac region. The Con A-treated embryos were confirmed to have Con A binding on the wall of the outflow tract in the cardiac region and in the mesenchyme of the cranial region, which are thought to be migration pathways of neural crest cells. These findings suggested that Con A inhibited the early migration of neural crest cells by binding directly to some substrates distributed along the pathways in the embryos, so that the neural crest cells could not punctually reach the locations where they would proliferate and differentiate into the destined cell types.