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Localization of CD 9 Molecule on Bull Spermatozoa: Its Involvement in the Sperm–Egg Interaction
Author(s) -
Antalíková J,
Jankovičová J,
Simon M,
Cupperová P,
Michalková K,
Horovská Ľ
Publication year - 2015
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.12508
Subject(s) - sperm , human fertilization , andrology , western blot , biology , monoclonal antibody , antibody , immunofluorescence , semen , microbiology and biotechnology , chemistry , immunology , anatomy , biochemistry , medicine , gene
Contents Tetraspanin CD 9 is one of the egg membrane proteins known to be essential in fertilization process. The presence and localization of CD 9 molecule in spermatozoa and its possible function in reproduction are still unclear. In our study, we describe the localization of CD 9 on bull spermatozoa. In the immunofluorescence assay, the positive signal has been observed in the high proportion of sperm cells as a fine grains either on the apical part or through the entire anterior region of sperm head. CD 9 recognized by monoclonal antibody IVA ‐50 was detected on freshly ejaculated (83.4 ± 3.7%) and frozen‐thawed (84.3 ± 2.3%) sperm. The same reaction pattern was observed on sperm capacitated for 1 h, 2 h, 3 h and 4 h (83.6 ± 2.0%; 84.0 ± 1.5%; 85.7 ± 0.8%; 77.5 ± 10.8%). The presence of CD 9 exclusively on plasma membrane of the bovine sperm has been detected by Western blot analysis of the protein fractions after the discontinuous sucrose gradient fractionation of the bull sperm. Moreover, probable role of the sperm CD 9 molecule in fertilization process of cattle has been suggested as sperm treatment with anti‐ CD 9 antibody significantly reduced (by 25%, p ≤ 0.001) the number of fertilized oocytes compared to control group in fertilization assay in vitro .