CONCURRENT ULTRASTRUCTURAL ANALYSIS AND GENOMIC ASSESSMENT OF SPERMATOZOA TO ACCURATELY DIAGNOSE RARE AND SEVERE FORMS OF MALE INFERTILITY

In men with ART failure, sperm DNA sequencing ultrastructural analysis can identify specific defects and guide treatment. the past 2 years, 22 consenting who failed IVF cycles due to severe astheno-/teratozoospermia were recruited; specimens from 3 fertile used as controls. To assess ultrastructure by transmission electron microscopy (TEM), pelleted spermatozoa fixed, embedded, sectioned 100-nm slides ultramicrotome, viewed an microscope (JEOL-1400) at 300,000X magnification. Sperm head morphology, fibrous sheath intactness, axonemal/periaxonemal structure examined on least 100 diagnose organelle defects. was extracted amplified 500 for next-generation (NGS) mutations contributing abnormalities. Four types of identified: globozoospermia, dysplasia (DFS), proximal centriole defect, primary ciliary dyskinesia (PCD). One case combined globozoospermia DFS one defect identified. 13 globozoospermic cases, 97–100% displayed complete spherical heads absence acrosomes, perinuclear theca deformities, dispersed chromatin. Microtubular arrangement centrosome flagellum conserved. NGS identified deletions (DPY19L, PICK1, SPATA16) directly responsible phenotype. Based findings, assisted oocyte activation performed 7 couples oocyte-activating agent, boosting fertilization 5.7% 40.2% (p<0.00001), resulting in 5 pregnancies 4 deliveries. patients (n=6), flagellar deformities observed 90–100% mitochondria disorganization, disarranged microtubules, cytoplasmic remnant enveloping coiled flagella, indicating incomplete late spermiogenesis. Normal acrosomes nuclei seen. (AKAP4, SPAG16, CATSPER1) involved development function. For (n=2), appeared normal. Approximately 90% centrioles exhibited microtubular confirmed ODF2 mutation NGS. a patient PCD, chaotic outer dynein arms prevailed axonemal cross-sections examined, DNAH5 gene deletion. Nonetheless, few motile ICSI, yielding 80% delivery healthy twin. Concurrent genomic pinpoint disruption lead successful cycles.