Evidence for the existence of juvenile hormone in the horseshoe crab

"Lipid-based hormones known as the juvenile hormones (JH) are ubiquitous among the arthropods, but their presence, functions, and sites of production in the horseshoe crab, Limulus polyphemus, remain unknown. Large size and lack of secondary sex characteristics in adult female horseshoe crabs may indicate continuous growth and molting throughout life, which is the outcome of high JH levels in insects and crustaceans. Here a study was undertaken to detect and localize lipid-based hormones in horseshoe crab hemolymph and tissue. Capillary electrophoresis and RP-HPLC analyses indicate the presence of a JH-like compound in subadult horseshoe crab hemolymph. The compound is present only in much lower amounts in the hemolymph of adult male and adult female horseshoe crabs. Identification of this compound was based on its similar retention time to standard JH, co-migration with added JH, and cross-reactivity with a polyclonal antibody to JH III. In addition, immunohistochemistry was used to localize the production site of this compound. Analysis of neural tissue, the assumed site of production, yielded no reactivity with labeled anti-JH III antiserum. In larval animals, however, reactivity was noted in yolk contained within the digestive tract. Since the larvae are lecithotrophic and feeding only on their yolk reserves, JH in the gut may be maternal, deposited in the egg before laying. Based on these results, we conclude that horseshoe crabs produce a lipid-based, JH-like hormone, with functional similarity to JH III in insects (i.e., maintenance of the juvenile form during growth and molting.) This paper is the first substantiation of such a hormone in horseshoe crabs. Our findings suggest that JH will be found in other chelicerates as well."