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Related Experiment Videos

Cloning, structure, and function of two rainbow trout Bf molecules

J O Sunyer1, I Zarkadis, M R Sarrias

  • 1Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia 19104-6079, USA.

Journal of Immunology (Baltimore, Md. : 1950)
|October 21, 1998
PubMed
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Rainbow trout possess two distinct factor B molecules, Bf-1 and Bf-2, involved in complement activation. Trout Bf-2 uniquely restores both alternative and classical complement pathways, offering insights into the evolution of these immune genes.

Area of Science:

  • Immunology
  • Evolutionary Biology
  • Genetics

Background:

  • Factor B (Bf) and C2 are linked MHC class III genes, likely originating from a single ancestral gene via duplication.
  • The evolutionary origin of this duplication event across animal phyla remains unknown.
  • Previous studies in zebrafish and medaka showed only one Bf/C2-like molecule, unlike mammals.

Purpose of the Study:

  • To characterize two novel factor B molecules (Bf-1 and Bf-2) in rainbow trout.
  • To investigate their roles in complement activation pathways.
  • To explore the evolutionary implications of multiple factor B genes in teleost fish.

Main Methods:

  • Molecular cloning and sequencing of Bf-1 and Bf-2 cDNAs.
  • Analysis of deduced amino acid sequences for similarity to mammalian Bf and C2.

Related Experiment Videos

  • Gene expression analysis (liver tissue).
  • Biochemical assays to assess C3 convertase formation and C3 cleavage.
  • Functional assays using depleted serum to evaluate hemolytic activity restoration.

    • Main Results:

    • Rainbow trout possess two Bf genes, Bf-1 and Bf-2, with 75% amino acid identity, more similar to mammalian Bf than C2.
    • Both trout Bf proteins can form the alternative pathway C3 convertase and undergo cleavage into Ba- and Bb-like fragments.
    • Trout Bf-2 demonstrated the remarkable ability to restore hemolytic activity in both alternative and classical complement pathways of depleted trout serum.

    Conclusions:

    • Rainbow trout have duplicated factor B genes, suggesting a different evolutionary trajectory compared to other teleosts like zebrafish and medaka.
    • Trout Bf-2 plays a crucial role in both major complement activation pathways, highlighting functional divergence or conservation.
    • These findings provide new insights into the evolution of the complement system and the MHC class III region in vertebrates.