The sperm membrane undergoes extensive surface remodeling as it matures in the epididymis. During this process, the sperm is encapsulated in an extensive glycocalyx layer, which provides the membrane with its characteristic negative electrostatic charge. In this study, we develop a method of microelectrophoresis and standardize the protocol to isolate sperm with high negative membrane charge. Under an electric field, the percentage of positively charged sperm (PCS), negatively charged sperm (NCS), and neutrally charged sperm was determined for each ejaculate prior to and following density gradient centrifugation (DGC), and evaluated for sperm DNA damage, and histone retention. Subsequently, PCS, NCS, and neutrally charged sperm were selected using an ICSI needle and directly analyzed for DNA damage. When raw semen was analyzed using microelectrophoresis, 94 % were NCS. In contrast, DGC completely or partially stripped the negative membrane charge from sperm resulting PCS and neutrally charged sperm, while the charged sperm populations are increased with an increase in electrophoretic current. Following DGC, high sperm DNA damage and abnormal histone retention were inversely correlated with percentage NCS and directly correlated with percentage PCS. NCS exhibited significantly lower DNA damage when compared with control (P < 0.05) and PCS (P < 0.05). When the charged sperm population was corrected for neutrally charged sperm, sperm DNA damage was strongly associated with NCS at a lower electrophoretic current. The results suggest that selection of NCS at lower current may be an important biomarker to select healthy sperm for assisted reproductive treatment.

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Optimization of microelectrophoresis to select highly negatively charged sperm.

Sperm selection is an important part of the ICSI and is based solely on sperm morphology and motility. The latter parameters may not be sufficient to select sperm with intact chromatin. Therefore, sperm selected based on sperm functional characteristics may result in selection of the most appropriate sperm for the ICSI procedure. In this chapter, we introduce two advanced sperm selection procedures based on electric charge on the sperm plasma membrane, Zeta method and electrophoretic method, and also discuss the importance of these methods for ART.

Sperm Selection Based on Surface Electrical Charge

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M Shayesteh Moghadam

Papers

The comparison of ha binding and zeta methods efficiency in selection of sperm with normal morphology and intact chromatin