New insights from the oyster Crassostrea rhizophorae on bivalve circulating hemocytes

PLoS One. 2013;8(2):e57384. doi: 10.1371/journal.pone.0057384. Epub 2013 Feb 25.

Abstract

Hemocytes are the first line of defense of the immune system in invertebrates, but despite their important role and enormous potential for the study of gene-environment relationships, research has been impeded by a lack of consensus on their classification. Here we used flow cytometry combined with histological procedures, histochemical reactions and transmission electron microscopy to characterize the hemocytes from the oyster Crassostrea rhizophorae. Transmission electron microscopy revealed remarkable morphological characteristics, such as the presence of membranous cisternae in all mature cells, regardless of size and granulation. Some granular cells contained many cytoplasmic granules that communicated with each other through a network of channels, a feature never previously described for hemocytes. The positive reactions for esterase and acid phosphatase also indicated the presence of mature cells of all sizes and granule contents. Flow cytometry revealed a clear separation in complexity between agranular and granular populations, which could not be differentiated by size, with cells ranging from 2.5 to 25 µm. Based on this evidence we suggest that, at least in C. rhizophorae, the different subpopulations of hemocytes may in reality be different stages of one type of cell, which accumulates granules and loses complexity (with no reduction in size) as it degranulates in the event of an environmental challenge.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Flow Cytometry
  • Hemocytes / cytology*
  • Microscopy, Electron, Transmission
  • Ostreidae / cytology*

Grants and funding

Conselho Nacional de Desenvolvimento Científico e Tenológico (CNPq), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Fundação Universitária José Bonifácio/Universidade Federal do Rio de Janeiro, Brazil. M.F.R., A.N. and S.A. are CNPq research fellows. R.M.M. holds a post-doctoral fellowship from PAPDRJ (FAPERJ/CAPES). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.