Quantification of proteomic profile changes in the hemolymph of crayfish during in vitro coagulation

Dev Comp Immunol. 2023 Oct:147:104760. doi: 10.1016/j.dci.2023.104760. Epub 2023 Jun 16.


Hemolymph is the circulatory fluid that fills the body cavity of crustaceans, analogous to blood in vertebrates. Hemolymph coagulation, similar to blood clotting in vertebrates, plays a crucial role in wound healing and innate immune responses. Despite extensive studies on the clotting process in crustaceans, no comparative quantitative analysis of the protein composition of non-clotted and clotted hemolymph in any decapod has been reported. In this study, we used label-free protein quantification with high-resolution mass spectrometry to identify the proteomic profile of hemolymph in crayfish and quantify significant changes in protein abundances between non-clotted and clotted hemolymph. Our analysis identified a total of two-hundred and nineteen proteins in both hemolymph groups. Furthermore, we discussed the potential functions of the top most high and low-abundant proteins in hemolymph proteomic profile. The quantity of most of the proteins was not significantly changed during coagulation between non-clotted and clotted hemolymph, which may indicate that clotting proteins are likely pre-synthesized, allowing for a swift coagulation response to injury. Four proteins still showed abundance differences (p < 0.05, fold change>2), including C-type lectin domain-containing proteins, Laminin A chain, Tropomyosin, and Reverse transcriptase domain-containing proteins. While the first three proteins were down-regulated, the last one was up-regulated. The down-regulation of structural and cytoskeletal proteins may affect the process of hemocyte degranulation needed for coagulation, while the up-regulation of an immune-related protein might be attributed to the phagocytosis ability of viable hemocytes during coagulation.

Keywords: Clot proteomics; Decapods; Innate immunity; Protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Astacoidea* / physiology
  • Blood Coagulation / physiology
  • Blood Coagulation Factors / metabolism
  • Hemocytes
  • Hemolymph* / metabolism
  • Proteomics


  • Blood Coagulation Factors