Relationship of immunohistochemical distribution of glycodelin and DNA fragmantation with the sperm morphology and their correlation with the patient's clinic

In reproductive medicine, sperm DNA integrity and capacitation mechanisms are considered to be important factors that affect both fertilization and embryo development. In this study, our aim was to observe DNA fragmentation and associated proteins and Glycodelin, which play important roles in capatitation, using immunohistochemistry in sperm smears and to evaluate correlation between clinical findings of patients with molecular changes.Sperm smears were prepared from 100 volunteer patients (aged 23 - 47) who presented to Andrology laboratory of Ege University Family Planning-Infertility Research and Treatment Centre and gave semen samples. History was taken about ages, job, post medical-family history, smoking, alcohol consumption. After 3-5 days of sexual abstinence, sperm count, sperm motility, leucocyte count, swim up scores of semen samples were evaluated with Makler counting chamber. Sperm smears were analyzed morphologically according to Kruger?s strict criteria. DNA fragmentation was evaluated by TUNEL (Terminal deoxyribonucleotidyl transferase mediated dUTP Nick?End Labeling), immunoreactivities of p53, Bcl-x, Fas, which play important roles in apoptotic process and immunoreactivity of Glycodelin S, transmembrane protein, were evaluated by avidin-biotin peroxidase method. Intensities of positive staining cells were determined as mild, moderate or severe and H-score results were obtained. TUNEL positive cells were expressed as per cent. The data were imput SPSS 15.0 and Mann Whitney U statistical test was used to compare the clinical findings.The number of TUNEL positive stained cells increased in the patient samples that had abnormal morphology, low sperm count and motility. Correlation was determined with increased immunoreactivities of p53 and Fas, especially in sperms which had head abnormality. While the immunoreactivity of Bcl-X was evaluated as mild in sperms that had normal morphology, sperms that had abnormal morphology was observed as having mild/moderate staining. While the immunoreactivity of Glycodelin was mild in small head and immature sperms, moderate immunoreactivity was seen in sperms which had normal morphology. Increased immunoreactivity was seen in other abnormalities.It was observed that patients past operations and affected the statistical sperm counting and motility. These patients had increased TUNEL positive cells. Patients who had normal morphology sperm the TUNEL positive cell counting was decreased .Clinical-pregnancy outcome is affected by good sperm selection for Intracytoplasmic Sperm Injection (ICSI) on morphological aspects by using high magnification microscope, strict criterias basis, sperm count, chromatin condensation and capacitation. Sperm DNA fragmentation , Glycodelin analysis is potentially valuable diagnostic and prognostic marker as compared to routine semen analysis in revealing paternal origin of some unexplained repeated ICSI failures in terms of fertilization and implantation failures.