Representation of an immune responsive gene family encoding fibrinogen-related proteins in the freshwater mollusc Biomphalaria glabrata, an intermediate host for Schistosoma mansoni.

Fibrinogen-related proteins (FREPs) are found in the hemolymph of the freshwater snail Biomphalaria glabrata, are up-regulated following exposure to digenetic trematode parasites, and bind to trematode larval surfaces, suggestive of a role in internal defense. Southern blot and degenerate-polymerase chain reaction (PCR) analyses were undertaken to better understand the diversity of the FREP-encoding gene family. Probes corresponding to the N-terminal IgSF domains of specific FREP gene subfamilies (FREPs 2, 3, 4, 7, 12 and 13) revealed between 1 to 8 loci per subfamily on Southern blots. Probes representing the relatively conserved C-terminal fibrinogen domain of FREPs bound many sequences in Southern blots of genomic DNA from B. glabrata, and from two related gastropod species, Biomphalaria pfeifferi and Helisoma trivolvis. Using degenerate-PCR, we obtained 42 unique fibrinogen-encoding sequences from 180 clones derived from a single individual of the M-line strain of B. glabrata, further supporting the notion of their abundant representation in the B. glabrata genome. The fibrinogen-encoding sequences of FREPs encoding one or two IgSF domains tended to separate into distinct clades, but bootstrap support for this separation was low. A novel category of fibrinogen-encoding sequence was also revealed. This study provides the approximate number of gene copies in several FREP subfamilies, confirms the existence of a diverse FREP gene family, reports additional unusual sequences encoding fibrinogen-like molecules, and provides further justification to explore the functional roles of FREPs in both B. glabrata and B. pfeifferi, both important intermediate hosts of the human pathogen, Schistosoma mansoni.