The Study Of Sperm Ultrastructure Of Male Patients With Unknown Infertility

Semen samples from unexplained infertility patients were selected as research materials, using transmission electron microscope to observe the distortion of the sperm ultrastructure,to explore the relationship between the abnormal ultrastructure and male infertility. Compare the ultrastructural differences between the head vacuoles in the sperm embeded with normal TEM methods and that in which only with glutaraldehyde fixation, result in the composition analysis in sperm head vacuoles.Experiment one: Two years of marriage, the woman with normal procreation, 8 cases of unexplained infertility patients and 3 cases of normal fertile men with one year marriage and give birth to children. After abstinence in 3 to 7 days, semen samples were collected by masturbation at room temperature. After liquefied, a part of semen are slowly injected into the centrifuge tube, with 650 g centrifuge for 10 min, abandon the upper seminal plasma after precipitation, using agarose to embed the sperm first, then fixation with glutaraldehyde about 1h, rinse with PBS(pH = 7.2) for 3 times(10 min/times), fixed with osmium tetroxide for 1 h, and then embed the materials with Epon812, all ultra-thin sections were stained with uranium and citrate, observation under transmission electron microscopy, mainly focusing on the ultrasructural changes in sperm acrosome, nucleus, mitochondria and microtubules etc.Experiment two:Vacuoles were commonly found in sperm head,the vacuoles' size are closely related with the function of acrosome, chromatin condensation and sperm DNA integrity,For further research the ultrastructural distortion of sperm head vacuoles in unexplained infertility patients and the vacuole composition, for the 8 cases of unexplained infertility patients and the 3 cases of normal reproductive male, we compare the ultrastructural differences between the head vacuoles, nucleus and acrosome in the sperm embeded with normal TEM methods and that in which only with glutaraldehyde fixation, result in the composition analysis in sperm head vacuoles.1.Through transmission electron microscope, the normal sperm plasma membrane and acrosome membrane structure is complete and clear, sperm nuclear presents pear-shaped, chromatin appears density, mitochondria in the tail are in the same size and morphology, arrayed rule, the microtubules �9+2' structure of the sperm tail remain intact.2.Infertility patients with abnormal sperm head, disordered ultrastructure of acrosome, reduced mitochondria or with uneven distribution and the turbulence of axial filament 9 + 2 form.3.The sperm study with IMSI observation and ultrastructural analysis showed that the abnormal sperm is higher in unexplained infertility patients than that in normal fertile male, and there is protein and lipid in the sperm head vacuoles.We found the ultrastructural changes in the sperm acrosome, sperm nucleus, mitochondria, and microtubules in unexplained infertility patients, always with the sperm head vacuoles increased. The ultrastructural changes and the increased sperm head vacuoles may be one od the reasons of male infertility, suggestting that unexplained infertility patients should be added the TEM Sperm Checking item, choosing sperm without or with less or small sperm head vacuoles used to IMSI(Intracytoplasmic Morphologically Selected Sperm Injection).