An immunological method to screen sex-specific proteins of bovine sperm
| Autor: | Liguo Yang, Guohua Hua, Jia-Jun Xiong, Aixin Liang, Lei Sang, Li Han, Li-Jun Huo, Wu-Cai Yang |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Male
endocrine system X Chromosome Immunoprecipitation Sexing Antibodies Flow cytometry Bovine sperm Y Chromosome Genetics medicine Animals Sex Preselection reproductive and urinary physiology Antiserum medicine.diagnostic_test biology urogenital system Immune Sera Membrane Proteins Rabbit (nuclear engineering) Spermatozoa Sperm Molecular biology biology.protein Cattle Animal Science and Zoology Rabbits Antibody Food Science |
| Zdroj: | Journal of Dairy Science. 94:2060-2070 |
| ISSN: | 0022-0302 |
| Popis: | This study was designed to identify sex-specific antibodies (SSAb) in rabbit antisera against bovine sex-sorted sperm, and capture sex-specific proteins of bovine X- or Y- proteins by SSAb. The rabbit antisera against bovine X- or Y-sperm were first produced by a series of immunological approaches, and further purified through immuno-neutralization with excess sex-sorted Y- or X-sperm, respectively, to remove non-sex specific antibodies and enrich sex-specific antibodies. After removal of non-sex specific antibodies, the purified rabbit sera with enriched sex-specific antibodies were screened for sex-specific antibodies by immunofluorescence staining and flow cytometry. The results showed that 3.0, 2.2, and 4.2% of unsorted sperm, sex-sorted X-sperm, and sex-sorted Y-sperm were recognized by the purified rabbit antisera against Y-sperm, respectively, whereas 29.2, 19.7, and 3.9% of unsorted sperm, sex-sorted X-sperm, and sex-sorted Y-sperm were recognized by the purified rabbit antisera against X-sperm. These results suggested that the purified rabbit antisera against X-sperm contained SSAb that preferentially bound to sex-sorted X-sperm. Subsequently, the purified rabbit antisera against X- or Y-sperm were used to immunoprecipitate sex-specific proteins in bovine sperm proteins, and a 30-kDa protein was specifically captured by the rabbit antisera against X-sperm. In conclusion, our results implied that this 30-kDa protein might be a sex-specific protein in bovine X-sperm, which has the potential to be used in immunological procedures for sexing sperm. |
| Databáze: | OpenAIRE |
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