8-Hydroxyguanine (8-oxoG) is the most common oxidative DNA lesion and unrepaired 8-oxoG is associated with DNA fragmentation in sperm. However, the molecular effects of 8-oxoG on spermatogenesis are not entirely understood. Here, we identified one infertile bull (C14) due to asthenoteratozoospermia. We compared the global concentration of 8-oxoG by reverse-phase liquid chromatography/mass spectrometry (RP-LC/MS), the genomic distribution of 8-oxoG by next-generation sequencing (OG-seq), and the expression of sperm proteins by 2-dimensional polyacrylamide gel electrophoresis followed by peptide mass fingerprinting (2D-PAGE/PMF) in the sperm of C14 with those of a fertile bull (C13). We found that the average levels of 8-oxoG in C13 and C14 sperm were 0.027% and 0.044% of the total dG and it was significantly greater in infertile sperm DNA (p = 0.0028). Over 81% of the 8-oxoG loci were distributed around the transcription start site (TSS) and 165 genes harboring 8-oxoG were exclusive to infertile sperm. Functional enrichment and network analysis revealed that the Golgi apparatus was significantly enriched with the products from 8-oxoG genes of infertile sperm (q = 2.2 × 10-7). Proteomic analysis verified that acrosome-related proteins, including acrosin-binding protein (ACRBP), were downregulated in infertile sperm. These preliminary results suggest that 8-oxoG formation during spermatogenesis dysregulated the acrosome-related gene network, causing structural and functional defects of sperm and leading to infertility.

UNASSIGNED: Assessment of relationship between LC3II/LC3 and Autophagy-related 7 (Atg7) proteins, as markers of autophagy, as well as evaluating the sperm parameters and DNA fragmentation in spermatozoa of infertile men with globozoospermia.
UNASSIGNED: In this case-control study, 10 semen samples from infertile men with globozoospermia and 10 fertile individuals were collected, and the sperm parameters, sperm DNA fragmentation, and main autophagy markers (Atg7 and LC3II/LC3) were assessed according to World Health Organization (WHO) criteria, TUNEL assay, and western blot technique, respectively.
UNASSIGNED: The mean of sperm concentration and motility were significantly lower, while the percentage of abnormal spermatozoa and DNA fragmentation were significantly higher in infertile men with globozoospermia compared to fertile individuals (P<0.01). Unlike the relative expression of LC3II/LC3 that did not significantly differ between the two groups, the relative expression of ATG7 was significantly higher in infertile men with globozoospermia compared to fertile individuals (P <0.05). There was a significantly negative correlation between the sperm concentration (r=-0.679; P=0.005) and motility (r=-0.64; P=0.01) with the expression of ATG7, while a significantly positive association was founf between the percentage of DNA fragmentation and expression of ATG7 (0.841; P =0.018).
UNASSIGNED: The increased expression of ATG7 and unaltered expression of LC3II/LC3 may indicate that the autophagy pathway is initiated but not completely executed in spermatozoa of individuals with globozoospermia. A significant correlation of ATG7 expression with increased sperm DNA fragmentation, reduced sperm concentration, and sperm motility may associate with the activation of a compensatory mechanism for promoting deficient spermatozoa to undergo cell death by the autophagy pathway. Therfore, this pathway could act as a double-edged sword that, at the physiological level, is involved in acrosome biogenesis, while, at the pathological level, such as globozoospermia, could act as a compensatory mechanism.

The acrosome is an organelle unique to sperm cells, thought to be indispensable for fertilization. In globozoospermia the main defect is characterized by the absent or severely malformed acrosome. Males with severe globozoospermia, to have the ability to father their own children, are committed to one of the most advanced and expensive method of assisted conception: in vitro fertilization with intracytoplasmic sperm injection (ICSI). However, rates of successful ICSI fertilization remain still very poor because globozoospermic spermatozoa have not the ability to trigger oocyte activation. It is emerging that the acrosome is not only fundamental for the release of lytic content to penetrate oocyte\'s coats; the strategies of oocyte penetration and activation by mammalian spermatozoa uncover less conventional and unexplored roles for acrosomal components. Discovered as cellular organelle in the early 1920s [1], the nature of the acrosome and its biogenesis are still matter of debate. In this review, I discuss the distinct classifications proposed in the years for the acrosome, providing also a glance on the intimate relationship that exists between acrosomal constituents and function. Emphasis is particularly devoted to the recent work on the acrosome biogenesis.

In the present study, the quality of frozen-thawed epididymal and testicular sperm recovered from a Siamese Eld\'s deer was examined. The epididymal sperm quality was assessed in fresh, cold-stored at 4°C and frozen-thawed samples. Zona binding ability of the frozen-thawed epididymal samples with Burmese Eld\'s deer oocytes was also evaluated. Testicular sperm extracted from tissues frozen at -80 or -196°C for one month were examined for membrane and DNA integrity. Epididymal sperm retained their quality for up to 24 hr of cold storage at 4°C. The percentages of sperm motility, intact membrane, intact acrosome and intact DNA were 30, 46.5, 27 and 89.5% in the frozen and thawed epididymal sperm, and the average ability to bind with oocytes was 92.5 ± 64 sperm/oocytes. Around 70% of the sperm extracted from testicular tissues cryopreserved at -196 and -80°C for one month showed an intact membrane. In conclusion, epididymal and testicular sperm survived for more than 13 hr post-mortem. Furthermore, cold storage at 4°C and cryopreservation at -196 and -80°C maintain the quality of epididymal and testicular sperm. This study represents a model for male gamete rescue in endangered Eld\'s deer.

OBJECTIVE: To determine if fertilization and embryo cleavage can be achieved by artificial oocyte activation in circumstances of repeated failed fertilization with sperm that have an acrosome.
METHODS: A woman with three IVF cycles with intracytoplasmic sperm injection (ICSI) failed to fertilize any eggs. The sperm had severe oligoasthenoteratozoospermia with no sperm with normal morphology. In the fourth IVF cycle fertilization was evaluated by performing ICSI with the husband\'s sperm and egg activation with calcium ionophore, ICSI with the husband\'s sperm without artificial oocyte activation, and ICSI with donor sperm.
RESULTS: Five mature oocytes were retrieved. Of the four eggs having ICSI with the husband\'s sperm only one of the two activated by calcium ionophore fertilized and resulted in a cleaved day 3 embryo. Interestingly, the one egg fertilized by donor sperm did not fertilize.
CONCLUSIONS: The data could be consistent with conclusions that in some cases the failure to fertilize may be related to an oocyte activation factor/receptor problem in the oocyte that can be overcome by the use of calcium ionophore.

Sperm acrosome is known to play a role in the fertilization of the majority of animal species studied. As a general rule, the acrosome appeared as soon as the fertilization occurred out of aquaeous phase. The biochemical content of acrosome as well as its release mode could suggest it is a simple lysosome. But this would by pass its important morphogenic role in spermiogenesis. Its development is strongly linked to the development of the microtubules manchette system. Molecular data of animal mutagenesis contribute to the understanding of acrosome biogenesis mechanisms. Globozoospermia is a rare but severe human teratozoospermia, characterized by ejaculates entirely consisting of round-headed spermatozoa that lack an acrosome. It originates from a disturbed acrosome biogenesis. Recently, the genetic study of a familial globozoospermia led to highlight a homozygote mutation of the gene SPATA16, linked to the globozoospermic phenotype. This study contributes to the understanding of the mechanisms implied in human acrosome formation.

We report the successful outcome of intracytoplasmic sperm injection (ICSI) treatment in two siblings with familial globozoospermia. After controlled ovarian hyperstimulation and oocyte pick-up, retrieved oocytes were mechanically activated before ICSI and a fertilization rate of 33.3% was achieved in the first case. The second couple underwent ICSI without oocyte activation and a 9.1% fertilization rate was obtained. The transfer of two grade I embryos in the first couple and one grade I embryo in the second couple resulted in clinical pregnancies with healthy livebirths. It was concluded that the main problem of cases with globozoospermia is a low fertilization rate, and even though ICSI and oocyte activation can increase this rate it is not necessarily needed to achieve a pregnancy.

We report a case of oligoasthenoteratozoospermia in a 40 year-old patient with a familial history that revealed multiple cases of infertility and perinatal deaths. The patient\'s semen sample contained 2x10(6) spermatozoa/ml, with an overall progressively motile population of <5%. Cytological analysis revealed a teratozoospermia with 100% of abnormal macrocephalic sperm heads and an irregular acrosomal cap in 38% of cells. Moreover, 72% of spermatozoa carried multiple flagella (2-5). The midpiece was elongated and/or enlarged with cytoplasmic droplets in 15% of cells. The multiple anomalies index (MAI) was 3.3 (normal value = 1.6), reflecting the high incidence of spermatozoal morphological abnormalities in this patient. Ultrastructural analysis revealed the presence of 2 or 3 vacuolated nuclei per sperm head. The acrosome was abnormal and the chromatin, partially packaged, appeared rough. In some cases, a large amount of cytoplasm containing vacuoles was observed around the nucleus and the acrosome. The mitochondrial helix was disorganized. Chromosome analysis performed on blood cells revealed a normal karyotype. Three-colour fluorescence in-situ hybridization (FISH) analysis of 1148 spermatozoa showed 21.6% to be diploid, 62.4% triploid, 13.3% quadriploid and 2.7% hyperploid (<4n). In conclusion, we suggest that this case could result from a genetically induced spermiation failure, the origin of which is discussed.

In cases of severe teratozoospermia, the current morphological criteria used to assess chromosomal status is insufficient for the selection of spermatozoa for intracytoplasmic sperm injection (ICSI). Case histories are reported of four patients presenting 100% teratozoospermia, and the integrity of their individual chromosomal statuses is determined using a three-colour fluorescence in-situ hybridization (FISH) technique. Patient 1 presented shortened flagella syndrome, patient 2 globozoospermia, patient 3 spermatozoa with irregular acrosomes, and patient 4 macrocephalic spermatozoa with associated multiple flagella. Three-colour FISH analysis using chromosome X, Y and 1-specific probes showed that approximately 95% of the spermatozoa analysed from patients 1, 2 and 3 presented X,1 and Y,1 signals, X,Y ratios and aneuploidy/diploidy rates comparable with those observed in normal controls. In contrast, patient 4 showed a highly elevated Y to X sex ratio and a highly elevated aneuploidy/diploidy rate. Three-colour FISH analysis thus demonstrates an increased incidence of chromosomal abnormalities in association with macrocephalic spermatozoa. Moreover, the analysis shows that in patients affected with either globozoospermia, shortened flagella syndrome or a condition of abnormal acrosomal spermatozoa, no association exists between chromosomal status and phenotype. Since these patients display normal haploid, sex chromosome and aneuploidy status, ICSI can be conceivably offered as a treatment for their infertility.

A case of headless or decapitated spermatozoa of an infertile man is reported. The 33-year-old patient showed almost normal spermiogram except for a high percentage (more than 90%) of headless spermatozoa. Both hypoosmotic swelling and zona-free hamster egg tests showed that most headless spermatozoa appeared to be normal in their function as tails. The ultrastructure of the ejaculated spermatozoa was observed using a surface replica method and by conventional ultrathin sectioning. The headless spermatozoa were almost normal in structure except for the absence of the head. Tailless heads were rarely observed. Varieties of ultrastructural abnormalities were observed in spermatozoa heads. Among them, abnormalities in the nuclear membrane covering the posterior pole of the nucleus were prominent. The implantation fossa and the basal plate were not formed. Instead, the nuclear membrane with numerous nuclear pores was found in this region. The decapitation seemed to take place between the region of the basal plate and the proximal centriole.