OPTIMIZATION AND EVALUATION OF AN ACROSOME ASSAY

THE ABILITY TO MEASURE AN ACROSOME REACTION IN HUMAN SPERM MAY LEAD TO NEW DIAGNOSES AND THERAPY FOR MALE INFERTILITY. A NUMBER OF TECHNIQUES HAVE BEEN DESCRIBED, BUT NONE ARE ROUTINELY USED IN A CLINICAL SEMEN ANALYSIS. THE OBJECTIVE OF THIS RESEARCH IS TO OPTIMIZE THE CONDITIONSFOR AN INDIRECT IMMUNOFLUORESCENCE ASSAY USING EXISTING MONOCLONAL ANTIBODIES, T6 AND MHS-10, TO QUANTITATE ACROSOME-REACTED SPERM. ASSAYS WILL BE DEVELOPED FOR DETERMINING THE ACROSOME STATUS OF A FRESH EJACULATE AS WELLAS FOR MEASURING THE ACROSOME-INDUCIBLE POPULATION IN A SEMEN SPECIMEN. THE ASSAY WILL BE OPTIMIZED FOR MINIMAL DAY-TO-DAY AND DONOR-TO-DONOR VARIABILITY. THE INDIRECT IMMUNOFLUORESCENCE ASSAY WILL ALSO BE EVALUATED USING AN ACROSOME INDUCTION TECHNIQUE THAT HAS BEEN EXTENSIVELY CORRELATED WITH IN VITRO FERTILIZATION AND PREGNANCY RATES. IF AN ACROSOME-REACTED POPULATION CAN BE CONSISTENTLY DETECTED, THE ASSAY WILL BE MOVED INTO CLINICAL TRIALS AT BAYLOR COLLEGE OF MEDICINE TO EVALUATE SPERM FROM INFERTILE PATIENTS.