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Flynn, K.M., P.K. Schoff, and J.M. Holy, 1997. Molecular Analysis of Eurasian
Ruffe Testicular Antigens Identified by a Panel of Monoclonal Antibodies, University
of Minnesota-Duluth
Proceedings of the International Symposium on Biology and Management of Ruffe, March
21-23, 1997
Molecular Analysis of Eurasian Ruffe Testicular Antigens Identified by a
Panel of Monoclonal Antibodies
ABSTRACT
As a first step in the long-term effort to control reproduction of xenobiotic teleost fish
by molecular methods, studies identifying and characterizing sperm and egg surface
proteins involved in Eurasian ruffe fertilization have been initiated. So far, a panel of
monoclonal antibodies have been produced using ruffe sperm and testicular tissue as
antigen. Robertsonian mice were injected and boosted with preparations of
formaldehyde-fixed milt, as well as with whole macerated testis. Screening of hybridomas
employed immunofluorescence analysis of formaldehyde-fixed cryostat sections of testes,
and resulted in the isolation of a eight hybridoma cell lines. Based on immunolabeling
patterns, four groups of testicular antigens have been identified: 1) "sperm
surface" (surfaces of sperm are strongly labeled); 2) "punctate"
(one-to-a-few strongly labeled foci are labeled within the sperm cytoplasm); 3)
"painted" (the entire spermatogenic epithelium is labeled in broad, curvilinear
patterns); and 4) "lumenal" (granular aggregations of material in the lumens of
germinal cysts are strongly labeled). Western blot analysis has resulted in the
identification of polypeptide candidates for most of these antigens, ranging from Mr
27,000 to 91,000. A number of these antibodies exhibit cross-reactivity to cryostat
sections of sauger testis, and three cross-react with sections of Xenopus laevis (the
South African clawed toad) testis. One of these includes a "punctate" class of
antibody, which in Xenopus appears to label intermediate filaments. This suggests that an
intermediate filament-like protein is present in a non-filamentous form in ruffe sperm. In
terms of future tools with which to potentially disrupt reproductive function at the level
of fertilization, the antibodies to the sperm surface are of the most obvious interest.
Further characterization of these antibodies is in progress, including screening of
zebrafish (Danio rerio) cDNA libraries.
Contact: Jon M. Holy, Natural Resources Research Institute, University
of Minnesota, Duluth, MN 55812
Keywords: Ruffe, Basic_biology
Product Type: Publications, Conference_proceedings
User Type: General
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