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Localisation of clusterin in normal human sperm by immunogold electron microscopy
M Atlas-White1, B F Murphy, H W Baker
1University of Melbourne, Department of Obstetrics and Gynaecology, The Royal Women's Hospital, Carlton, Victoria, Australia.
Pathology
|February 24, 2001
Summary
A novel acrosomal form of clusterin is found within the acrosomal contents of intact human sperm. This specific localization was confirmed using immunogold electron microscopy, highlighting its association with sperm function.
Area of Science:
- Reproductive Biology
- Molecular Biology
- Cell Biology
Background:
- Two forms of clusterin exist in the male reproductive tract: a conventional heterodimeric form and a novel acrosomal form.
- The acrosomal clusterin is specifically detected on human sperm in the acrosomal region, but only in acrosome-intact sperm.
Purpose of the Study:
- To precisely determine the localization of the acrosomal form of clusterin within human sperm.
- To investigate the association of acrosomal clusterin with specific sperm structures.
Main Methods:
- Immunogold electron microscopy was employed on normal human sperm.
- The E5 anti-clusterin monoclonal antibody (mAb) and a pre-embedding technique were utilized.
- Spermatozoa were fixed with ethanol and permeabilized with Triton X-100 to expose internal structures.
Main Results:
- Acrosomal clusterin was localized to the acrosomal contents.
- Specific immunogold labeling was observed over the exposed acrosomal contents after plasma membrane and outer acrosomal membrane removal.
- Immunogold particles were also detected in the equatorial segment of the sperm.
Conclusions:
- The acrosomal form of clusterin is confirmed to be associated with the contents of the acrosome.
- This finding provides insights into the specific role and location of clusterin in sperm biology.