Many benthic marine invertebrates, like barnacles, have a planktonic larval stage whose primary purpose is dispersal. How these species colonize suitable substrata is fundamental to understanding their evolution, population biology, and wider community dynamics. Unlike larval dispersal, settlement occurs on a relatively small spatial scale and involves larval behavior in response to physical and chemical characteristics of the substratum. Biogenic chemical cues have been implicated in this process. Their identification, however, has proven challenging, no more so than for the chemical basis of barnacle gregariousness, which was first described >50 years ago. We now report that a biological cue to gregarious settlement, the settlement-inducing protein complex (SIPC), of the major fouling barnacle Balanus amphitrite is a previously undescribed glycoprotein. The SIPC shares a 30% sequence homology with the thioester-containing family of proteins that includes the âŁ2-macroglobulins. The cDNA (5.2 kb) of the SIPC encodes a protein precursor comprising 1,547 aa with a 17-residue signal peptide region. A number of structural characteristics and the absence of a thioester bond in the SIPC suggest that this molecule is a previously undescribed protein that may have evolved by duplication from an ancestral âŁ2-macroglobulin gene. Although the SIPC is regarded as an adult cue that is recognized by the cyprid at settlement, it is also expressed in the juvenile and in larvae, where it may function in larva-larva settlement interactions.arthropod Í larva Í pheromone Í settlement cue C harles Darwin (1) was among the first to appreciate the biological diversity of the cirripedes, whose evolution is likely to be influenced strongly by their settlement behavior. Barnacles that settle gregariously have since featured prominently in studies that have shed light on ecological processes in the marine environment (2). Some barnacle species are also major components of hard fouling communities on ship hulls and other artificial marine structures. After the recent ban on the application of organotin-containing antifouling coatings that were linked to major environmental problems (3), interest in the settlement behavior of these barnacles and other fouling organisms has intensified. Whether in a fundamental or applied context, there have been many attempts to characterize chemical cues that modulate the settlement behavior of marine invertebrate larvae. Such studies have generally met with limited success, and the reasons for this have been explained in detail by others (e.g., ref.
4).A century after Darwin (1), Knight-Jones and Crisp (5) proposed that a factor associated with the cuticle of adult barnacles induces gregarious settlement of cypris larvae. Exhaustive studies on temperate species, particularly Semibalanus balanoides, suggested that the inductive cue, which could be extracted from tissues with seawater and partially purified by boiling, was a polymorphic system of closely related, watersoluble glycoproteins. The cue was name...