This enzyme has been documented as a sperm factor or an oocyte-activating factor, necessary to induce Ca2+oscillations after fertilization (Yoonet al., 2008). This case study reports a comprehensive clinical approach to complete globozoospermia with analyses of ultrastructure, DNA fragmentation levels, aneuploidy rate, detection of PLC and ICSI outcome SGK2 (with and without the use of a calcium ionophore in sibling oocytes), resulting in a Necrostatin 2 pregnancy. == Materials and methods == A patient with known globozoospermia, age 28 years and in otherwise good health, was enrolled for ICSI. with calcium ionophore resulted in high rates of fertilization, and an ongoing pregnancy was established after transfer of cryopreservedthawed embryos. Keywords:globozoospermia, ICSI, oocyte activation, phospholipase C zeta, pregnancy == Introduction == Globozoospermia is a term used to describe spermatozoa with an absence of acrosomes. Globozoospermia was first documented in 1965 (Meyhfer, 1965). Later,Schirren and colleagues (1971)provided ultrastructural details of globozospermic spermatozoa with the important finding that these abnormal spermatozoa lacked acrosomes. During spermiogenesis, the acrosome is formed by the Golgi apparatus of the developing spermatocyte. After positioning of the prospective acrosomal cap, the sperm head is remodelled cytoskeletally to a species-specific shape. In patients with globozoospermia, this process does not occur, leaving spermatozoa with a characteristic round-head appearance and functionally with limited capacity to fertilize (Damet al., 2007). Incidence has been estimated to be less than 0.1% of infertility patients (Holsteinet al., 1973) and considerable evidence indicates that complete globozoospermia can be of genetic origin (Damet al., 2007). Prior to 1995, patients with complete globozoospermia were left Necrostatin 2 with no options other than the use of donor spermatozoa or adoption. With the advent of intracytoplasmic sperm injection (ICSI), globozoospermia became one of the many severe male factor conditions that could be effectively treated. In 1995, an initial report was published where globozoospermic spermatozoa were used for IVF augmented Necrostatin 2 with ICSI (Liuet al., 1995). Later reports indicated that induction of oocyte activation using a calcium ionophore was necessary after ICSI in some patients with globozoospermia (Tejeraet al., 2008), strongly pointing to an absence of a putative oocyte-activating factor. However, at least two reports in the literature demonstrated that calcium ionophore treatment was not necessary (Diricanet al., 2008). Sperm phospholipase C zeta (PLC) has recently been implicated in oocyte activation (Yoonet al., 2008). This enzyme has been documented as a sperm factor or an oocyte-activating factor, necessary to induce Ca2+oscillations after fertilization (Yoonet al., 2008). This case study reports a comprehensive clinical approach to complete globozoospermia with analyses of ultrastructure, DNA fragmentation levels, aneuploidy rate, detection of PLC and ICSI outcome (with and without the use of a calcium ionophore in sibling oocytes), resulting in a pregnancy. == Materials and methods == A patient with known globozoospermia, age 28 years and in otherwise good health, was enrolled for ICSI. Physical examination was unremarkable and the karyotype was 46XY, negative for Y chromosome microdeletions. His family history was negative and a brother had children that were conceived naturally. Genetic counselling was given to the patient regarding potential heritable globozoospermia in male offspring and treatment was approved by the institutional review board at Eastern Virginia Medical School. The patient collected four semen samples. The initial sample was used for semen analysis and TdT (terminal deoxynucleotidyl transferase)-mediated dUDP nick-end-labelling (TUNEL). A second sample was used for transmission electron microscopy. A third sample was split for semen analysis, TUNEL and immunoblotting and immunofluorescence for PLC. A fourth semen collection was evaluated for aneuploidy by fluorescence in-situ hybridization (FISH). A semen sample from a fertile, normozoospermic individual was processed as a control. == Semen analysis == Semen was collected by masturbation into a sterile specimen container. A portion of the initial semen sample was cultured forUreaplasma urealyticumusing 10B broth (Remel, Lenexa, KS, USA) with simultaneous culture on A8 agar (Hardy Diagnostics, Santa Maria, CA, USA). The semen was analysed according to WHO standards (WHO, 1999). The presence of leukocytospermia was evaluated by peroxidase staining. Strict criteria (Menveld,et al., 1990) were used for sperm morphology. Concentration and the percentage of motile spermatozoa were determined manually using Makler counting chambers in duplicate. Other motion parameters were analysed using CASA (Hamilton-Thorne Research, Beverly, MA) and manually monitored as appropriate (Oehningeret al., 1990). Morphology smears were stained using Stat III Andrology Stain (Mid-Atlantic Diagnostics, Mt Laurel, NJ, USA), a cytology staining system equivalent to Diff-Quik for strict criteria. Morphology slides were scanned at 400 magnification to verify the absence of acrosomes, with final evaluation at 1000 magnification. The liquefied semen was.