Recent studies have shown that infertility affects estimated 15% of all couples. Male infertility is the primary or contributory
cause in 60% of these cases. Consequently, the application of assisted reproduction is increasing. These methods could
benefit from an extended evaluation of sperm quality. For this reason, we analyzed sperm proteins from 30 men with normal
spermiograms and 30 men with asthenozoospermia. Ejaculates of both groups were tested by flow cytometry (FCM) and
fluorescence with a set of well‑characterized anti‑human sperm Hs‑monoclonal antibodies (MoAbs), which were generated in our
laboratory. No statistically significant differences were found between normospermics and asthenospermics in the expression of
the sperm surface protein clusterin, evaluated with Hs‑3 MoAb, and semenogelin, evaluated with Hs‑9 MoAb. However, FCM
revealed quantitative differences in the acrosomal proteins between normozoospermic and asthenozoospermic men, namely, in
glyceraldehyde‑3‑phosphate dehydrogenase, evaluated with Hs‑8 MoAb, valosin‑containing protein, evaluated with Hs‑14 MoAb,
and ATP synthase (cAMP‑dependent protein kinase II, PRKAR2A), evaluated with MoAb Hs‑36. Asthenozoospermic men displayed
a highly reduced expression of intra‑acrosomal proteins, with a likely decrease in sperm quality, and thus a negative impact on
successful reproduction. Asthenozoospermia seems to be a complex disorder involving intra‑acrosomal proteins.

Keywords: asthenozoospermia; flow cytometry; fluorescence microscopy; monoclonal antibodies; sperm proteins

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