Sperm selection in assisted reproduction: A review of established methods and cutting-edge possibilities

Glioma pathogenesis-related 1-like protein1 (GliPr1L1) was identified by liquid chromatography-tandem mass spectrometry analyses of proteins associated to bovine sperm lipid raft membrane domains. This protein belongs to the CAP superfamily including cysteine-rich secretory proteins, Antigen 5 and pathogenesis-related 1 protein. PCR analysis revealed that GliPr1L1 is expressed in testis and, at a much lower level, all along the epididymis. Western blotting showed a similar distribution of GliPr1L1 in testicular and epididymal tissue extracts. In the epididymal lumen, GliPr1L1 was associated with the maturing spermatozoa and epididymosomes all along the excurrent duct but was undetectable in the soluble fraction of epididymal fluid. The protein was detectable as multiple isoforms with a higher MW form in the testis and proximal caput. Treatments with PNGase F revealed that N-glycosylation was responsible of multiple bands detected on Western blots. These results suggest that the N-glycosylation moiety of GliPr1L1 is processed during the transit in the caput. Western blots demonstrated that GliPr1L1 was associated with the sperm plasma membrane preparation. GliPr1L1 is glycosyl phosphatidyl inositol (GPI) anchored to caput and cauda spermatozoa as demonstrated by the ability of phosphatidylinositol specific phospholipase C to release GliPr1L1 from intact sperm cells. Lipid raft membrane domains were separated from caput and cauda epididymal spermatozoa. GliPr1L1 was immunodetectable in the low buoyant density fractions where lipid rafts are distributed. GliPr1L1 was localized on sperm equatorial segment and neck. In vitro fertilization performed in presence of anti-GliPr1L1 showed that this protein is involved in sperm–zona pellucida interaction. J. Cell. Physiol. 227: 3876–3886, 2012. © 2012 Wiley Periodicals, Inc.

Bovine sperm raft membrane associated Glioma Pathogenesis-Related 1-like protein 1 (GliPr1L1) is modified during the epididymal transit and is potentially involved in sperm binding to the zona pellucida.

The aim of this study was to compare the effects of spermatozoa separation techniques on sperm quality and in-vitro fertilization (IVF) results for cryopreserved bovine semen. Sephadex, glass wool and Percoll gradient separation techniques were used for sperm separation and sperm motility, morphology and membrane integrity were evaluated before and after separation. Also, cleavage and blastocyst developmental rate were investigated after IVF with sperm recovered by each separation technique. The motility of samples obtained by the three separation techniques were greater compared to the control samples (p < 0.05). The percentage of spermatozoa with intact plasma-membrane integrity, identified by 6-carboxyfluoresceindiacetate/ propidium iodide fluorescent staining and the hypo-osmotic swelling test, was highest in the glass wool filtration samples (p < 0.05). The cleavage and blastocyst rate of total oocytes produced from glass wool filtration samples were also higher than the control and Sephadex filtration samples (p < 0.05), but were not significantly different from Percoll separation samples. However, a significantly greater number of cleaved embryos produced by glass wool filtration developed to blastocyst stage than those produced by Percoll separation (p < 0.05). These results indicate that spermatozoa with good quality can be achieved by these three separation techniques and can be used for bovine IVF. In particular, it suggests that glass wool filtration would be the most effective method of the three for improving sperm quality and embryo production for cryopreserved bovine spermatozoa.

/pdf/a-comparative-study-of-sephadex-glass-wool-and-percoll-7oow6ha0e2.pdf

A comparative study of Sephadex, glass wool and Percoll separation techniques on sperm quality and IVF results for cryopreserved bovine semen

The straight line velocity of 500 individual spermatozoa was measured immediately before their direct injection into the cytoplasm of 500 metaphase II oocytes. The straight line velocity (VSL) did not have a normal distribution (P < 0.01) and ranged between 0 and 35 microm/s. The fertilization rate (84%) was significantly (P < 0.008) higher in the quartile of the sperm population with the highest VSL compared to the fertilization rate (68%) in the quartile with the lowest VSL. Embryo cleavage and embryo quality were not different in the quartiles of the sperm population used for injection.

A first prospective study of the individual straight line velocity of the spermatozoon and its influences on the fertilization rate after intracytoplasmic sperm injection.

We investigated the possibility of ultrastructural damage to human spermatozoa induced by different sperm preparation techniques. Ejaculates from 20 normozoospermic men were divided into equal aliquots and processed by glass wool filtration, Percoll density gradient centrifugation, and a simple two-step centrifugation procedure which served as a control. The evaluation of 60 spermatozoa from each of 20 test subjects (in all, n = 1200) ensured that a sufficiently large number of spermatozoa were investigated. Ultrastructural damage was assessed by scanning electron microscopy. We investigated the state of the acrosome after sperm preparation and measured the percentage of intact spermatozoal structures compared with that of the control. Compared with Percoll density gradient centrifugation, glass wool filtration yielded a significantly increased proportion of intact acrosomes. However, both preparations gave significantly better results than the control. In conclusion, both glass wool filtration and Percoll centrifugation are efficient techniques for the accumulation of spermatozoa with intact acrosomes. Because of the significantly higher percentage of intact acrosomes, glass wool filtration appears to be the more appropriate method. The significance of the conspicuous bending of sperm tails after Percoll centrifugation is not yet known.

/pdf/glass-wool-filtration-leads-to-a-higher-percentage-of-3e8fwok5lg.pdf

Glass wool filtration leads to a higher percentage of spermatozoa with intact acrosomes: an ultrastructural analysis.

The characteristics of the glass wool filtration technique for the preparation of semen samples in an intracytoplasmic sperm injection programme (ICSI) were compared with a two-layer Percoll density gradient procedure. Half of each of 25 semen samples were prepared by each technique. The oocytes from each patient were randomly injected, half with spermatozoa prepared by glass wool filtration and half with Percoll-separated spermatozoa from the same semen sample. The percentage of recovered motile spermatozoa, the total motile sperm count, the percentage of morphologically normal forms, the fertilization rate, the cleavage rate and embryo quality obtained with both preparations were analysed. The 95% confidence intervals obtained through the Altman-Bland analysis showed that the percentage of motile spermatozoa recovered was 3.5-9.9% higher with the glass wool filtration technique, and this was highly correlated (r = 0.92, P = 0.0001) to the method. Similarly, the total number of spermatozoa available for ICSI was 0.18 x 10(6)-2.44 x 10(6) higher with the glass wool column and was highly correlated to the method (r = 0.956, P = 0.0001). Also, the percentage of normal forms was 1.25-3.31% higher after glass wool filtration but was poorly correlated to the method (r = 0.47, P = 0.017). Out of 100 metaphase II oocytes injected with glass wool-extracted spermatozoa, 77 fertilized and 72 cleaved. Out of 97 metaphase II oocytes injected with Percoll-selected spermatozoa, 71 fertilized and 69 cleaved. These results were not statistically different. The mean +/- SEM embryo quality score for the glass wool group (2.90 +/- 0.27) was the same as that for the Percoll group (2.80 +/- 0.24). No fertilization failures occurred and 11 patients (44% per oocyte retrieval) became pregnant. It was concluded that glass wool filtration for semen preparation in an ICSI programme offers higher sperm recovery and sperm morphology of superior quality than with the classic two-layer Percoll gradient method, without affecting the fertilization rate and embryo quality.

/pdf/glass-wool-column-filtration-an-advantageous-way-of-18pxp8ik67.pdf

Glass wool column filtration, an advantageous way of preparing semen samples for intracytoplasmic sperm injection: an auto-controlled randomized study.

Motivation SIA-BRA is a data set that compiles stable carbon (C) and nitrogen (N) isotope ratios of terrestrial and aquatic animals sampled in Brazilian biomes and coastal marine areas. Stable isotope ratios are helpful in animal ecology for several reasons; for instance, they can be used to investigate trophic niches, energy sources (diet tracing) and to track migration patterns. The Neotropics are considered one of the most undersampled regions of the world. Given that Brazil is a continental country where most of the dietary ecology of animal species is under-assessed, we believe that the SIA-BRA can provide important complementary information to address this gap in the literature. Additionally, the SIA-BRA data set allows future investigations to address many questions concerning diet tracing, habitat use, food webs, foraging ecology, physiological aspects and effects of phylogeny on dietary ecology. Main type of variable investigated Carbon and nitrogen stable isotope ratios for terrestrial and aquatic animals. Spatial location and grain The SIA-BRA included animal tissues sampled in 964 sites in the main Brazilian biomes and coastal marine areas. Time period The data represent information published and/or sampled from 1984 to 2021. Major taxa studied and measurement level The SIA-BRA contains isotopic data of c. 21,804 non-captive wildlife specimens, excluding livestock production or laboratory experiments. They were 13,881 vertebrates and 7,923 invertebrates. They were divided into the following habitats: terrestrial (30% of the total), freshwater (27%), oceanic (40%) and estuarine (4%). There were 11 phyla, with a clear dominance of Chordata (64%) and Arthropoda (29%), 36 classes, 154 orders, 473 families, 894 genera and 1,157 species. Software format Data are supplied as a comma-delimited text file (.csv). 

SIA‐BRA: A database of animal stable carbon and nitrogen isotope ratios of Brazil

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Glass wool column filtration, an advantageous way of preparing semen samples for intracytoplasmic sperm injection: an auto-controlled randomized study.

SIA‐BRA: A database of animal stable carbon and nitrogen isotope ratios of Brazil