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A testis-specific antigen of the C57BL/6 mouse
O Mikami1, H Sakamoto, Y Yamamoto
1Department of Genetics, Hyogo College of Medicine, Japan.
Journal of Reproductive Immunology
|May 1, 1989
Summary
Researchers developed a monoclonal antibody (Moab) targeting specific testicular cells. This Moab identified a testicular antigen (Moab1A1-TA) composed of a peptide and carbohydrate, with antigenicity localized to the carbohydrate epitope.
Area of Science:
- Immunology
- Reproductive Biology
- Biochemistry
Background:
- Monoclonal antibodies (Moabs) are crucial tools in biological research.
- Understanding the specific antigens involved in spermatogenesis is vital for reproductive health studies.
Purpose of the Study:
- To develop and characterize a Moab specific to testicular cells.
- To identify the biochemical nature of the recognized testicular antigen (Moab1A1-TA).
Main Methods:
- Hybridoma technology for Moab production.
- Indirect immunofluorescence for tissue specificity.
- ELISA, SDS-PAGE, and Western blotting for antigen analysis.
- Enzymatic digestion (proteolytic, peptidases, glycolytic, keratanase) to determine antigen composition.
Main Results:
- Two hybridomas producing IgM were generated.
- Moab 1A1 specifically recognized the cytoplasm of spermatogonia, spermatocytes, and spermatids.
- Moab1A1-TA, detected in the void volume, showed a broad molecular weight range (45-205 kD).
- Moab1A1-TA is composed of an acidic peptide and a carbohydrate molecule, with antigenicity residing in the carbohydrate epitope.
- The carbohydrate moiety contains a lacto-series structure.
Conclusions:
- Moab 1A1 is a specific marker for early-stage male germ cells.
- The testicular antigen (Moab1A1-TA) is a glycoprotein with a carbohydrate epitope responsible for antigenicity.
- The presence of a lacto-series structure suggests specific glycosylation patterns in testicular antigens.