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          Institute: MPI für Evolutionsbiologie     Collection: Evolutionary ecology     Display Documents



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ID: 327679.0, MPI für Evolutionsbiologie / Evolutionary ecology
Characterization of antisera raised against stickleback (Gasterosteus aculeatus) MHC class I and class II molecules
Authors:Scharsack, J. P.; Kalbe, M.; Schaschl, H.
Language:English
Date of Publication (YYYY-MM-DD):2007-11
Title of Journal:Fish & Shellfish Immunology
Volume:23
Issue / Number:5
Start Page:991
End Page:1002
Review Status:not specified
Audience:Not Specified
Abstract / Description:The three-spined stickleback (Gasterosteus aculeatus) is an important model organism for investigations on the maintenance of polymorphism of the major histocompatibility complex (MHC) of vertebrates. Analysis of functional aspects of MHC diversity in stickleback would benefit from the availability of MHC specific reagents. Here we characterize antisera raised against recombinant fusion proteins of stickleback MHC class I alpha and class II alpha and beta. Western blot analysis using recombinant proteins confirmed the specificity of the antisera. In brain and muscle preparations, neither of the MHC types was detectable. High levels of each MHC receptor type were observed in gills and spleen and lower levels in head kidneys. In histological sections of gills, epithelial cells of primary and secondary lamellae stained positive with MHC class I antiserum, while single, scattered cells stained positive for MHC class II. In sections of spleen and head kidney, considerable numbers of cells positive for either MHC type were detected. Molecular weight shift in SDS-PAGE after deglycosylation of MHC class I alpha and class II beta confirmed the predicted glyco-protein character of the molecules. The majority of MHC II alpha was not glycosylated; only a small fraction of MHC II alpha was susceptible to deglycosylation. This suggests differential expression of the two stickleback MHC II alpha genes (Gaac-DAA, Gaac-DBA) only one of which (Gaac-DBA) has a site for N-linked glycosylation.
Free Keywords:Gasterosteus aculeatus; MHC class I; MHC class II; ECL; N-glycosylation; western blot; immunohistology
External Publication Status:published
Document Type:Article
Affiliations:MPI für Limnologie/Abt. Evolutionsökologie
External Affiliations:Konrad Lorenz Institute for Ethology, Savoyenstrasse 1a, A-1160 Vienna, Austria
Identifiers:ISSN:1050-4648 [ID-No:1]
DOI:10.1016/j.fsi.2007.03.011 [ID-No:2]
LOCALID:2578/S 38732 [Listen-Nummer/S-Nummer]
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