Characterization of the thrombospondin (TSP)-II gene in Penaeus monodon and a novel role of TSP-like proteins in an induction of shrimp sperm acrosome reaction

Abstract

We have recently shown that water-soluble materials from the egg extracellular cortical rods (wsCRs) exert the ability to induce the sperm acrosome reaction in Penaeus monodon. In this study, we further demonstrated that the thrombospondin protein family (TSP) existed in wsCRs, and that their mRNA transcripts were detected in developing oocytes as early as stage I. Full sequence analysis revealed that our pmTSP sequence was considerably different from the recently reported pmTSP in the 5′ nonconserved region and in many TSP signature domains, hence, the name pmTSP-II was given to our variant. The transcripts of pmTSP-II were detected only in early developing oocytes (stage-I and -II) while TSP-like proteins were detected in all developing oocytes, particularly at the outer rim of cortical rods situated in the extracellular crypts of the mature, stage-IV oocytes. In addition, wsCRs contained anti-TSP-reactive proteins, suggesting that TSP-like proteins are dissolved in and are part of the egg water during spawning. The functional importance of TSP-like proteins was evident by the interference of a wsCR-induced acrosome reaction response with anti-TSP in a concentration-dependent manner. In summary, we found that pmTSP-II transcripts were present in the developing oocytes and pmTSP-II protein accumulated in cortical rods, which are partly secreted and thus solubilized to produce dissolved TSP-like proteins that participate in induction of the sperm acrosome reaction-a novel reproductive role for TSP protein family. © 2013 Wiley Periodicals, Inc.