Pathogenic missense variation in PABPC1L/EPAB causes female infertility due to oocyte maturation arrest at the germinal vesicle stage.

Women undergoing controlled ovarian hyperstimulation prior to in vitro fertilization (IVF) are treated using various protocols to induce multiple follicular growths. Complete failure of all oocytes to mature during IVF cycles is rare; however, it is a known cause of primary female infertility. Recently, pathogenic variations in a few genes have been identified in women with oocyte maturation defects; however, the underlying genetic causes remain largely unknown.This study included a Turkish family comprising three sisters with recurring oocyte maturation arrest at the germinal vesicle stage after multiple ovarian stimulations. Exome sequencing revealed a homozygous missense variant (c.1037C>T, p.Ala346Val) in the EPAB gene (also known as PABPC1L) in all three affected sisters, which was either absent or heterozygous in the unaffected family members. Functional experiments confirming the pathogenicity of the variant were performed by transfecting HEK293T cells and demonstrated the instability and increased rate of proteolysis of the mutated PABPC1L/EPAB protein. The identified variant, located in the well-conserved fourth RNA recognition motif (RRM4), in silico 3D modelling suggested changes in the physical properties of the pathogenic variant of PABPC1L/EPAB. Our findings validate PABPC1L/EPAB as an essential genetic contributor to the oocyte maturation process in humans and have direct implications for the genetic counselling of patients and their family members.

Evaluation of α5ß1 integrin as a candidate marker for fertility in bull sperm samples.

With the progressive increase in the use of reproductive biotechnologies in the cattle industry, like artificial insemination and in vitro embryo production, the accurate determination of fertilizing competence of cryopreserved sperm samples is an essential issue. The routine methodology to assess bull sperm quality relies primarily on count, viability and motility of spermatozoa. However, these parameters do not tightly predict the reproductive success of samples. Therefore, identification of complementary markers of sperm functionality to strengthen the predictability of traditional spermogram is desirable to improve livestock reproduction practices. Previous results from our laboratory indicated that α5ß1 integrin plays a key role in bovine sperm function and mediates their interaction with the female reproductive tract. Thus, this study aimed to investigate whether the localization of α5ß1 held a correlation with fertilizing ability of bovine cryopreserved semen samples. Firstly, we assessed the quality of samples from six different bulls (A-F). We determined motility and viability of sperm samples after thawing and selection. Additionally, we measured the capacitation state of the samples by chlortetracycline (CTC) assay in the presence or absence of heparin, as an indicator of their responsiveness to a capacitating stimulus. Based on these assays, samples were classified being A the bull with the lowest quality and F the bull with the highest quality. Then, we studied the presence and localization of α5ß1 integrin. This protein showed a distribution pattern in the acrosomal (A), post-acrosomal (P) and acrosomal + post-acrosomal (A + P) regions with different localization percentages among the studied samples. Next, we determined the fertilizing ability of the samples in in vitro fertilization (IVF) assays and performed correlation analyses between IVF outcome and the routine spermogram parameters or α5ß1 integrin localization patterns. When the percentage of cells showing α5ß1 integrin was compared to fertilization rate, no correlation was observed. However, the presence of α5ß1 integrin in P and A + P regions (PA pattern), positively correlated with IVF rate (p < 0.05). These results suggest that while routine semen analyses failed to predict sperm reproductive competence, integrin localization in post-acrosomal region (PA pattern) showed a positive correlation with IVF outcome, thus posing an attractive marker to predict more accurately the reproductive performance of an individual.

MRP4-mediated cAMP efflux is essential for mouse spermatozoa capacitation.

Mammalian spermatozoa must undergo biochemical and structural changes to acquire the capacity for fertilization, in a process known as capacitation. Activation of PKA enzymes is essential for capacitation, and thus cAMP levels are tightly regulated during this process. Previously, we demonstrated that during capacitation, bovine spermatozoa extrude cAMP through multidrug resistance-associated protein 4 (MRP4, also known as ABCC4), which regulates intracellular levels of the nucleotide and provides cAMP to the extracellular space. Here, we report the presence of functional MRP4 in murine spermatozoa, since its pharmacological inhibition with MK571 decreased levels of extracellular cAMP. This also produced a sudden increase in PKA activity, with decreased tyrosine phosphorylation at the end of capacitation. Blockade of MRP4 inhibited induction of acrosome reaction, hyperactivation and in vitro fertilization. Moreover, MRP4 inhibition generated an increase in Ca2+ levels mediated by PKA, and depletion of Ca2+ salts from the medium prevented the loss of motility and phosphotyrosine inhibition produced by MK571. These results were supported using spermatozoa from CatSper Ca2+ channel knockout mice. Taken together, these results suggest that cAMP efflux via MRP4 plays an essential role in mouse sperm capacitation.This article has an associated First Person interview with the first author of the paper.

Sciatic nerve compression related to ossification of the sacrospinous ligament secondary to pelvic balance abnomalities.

The authors report an isolated case of right sacrospinous ligament ossification causing sciatic pain because of compression of the proximal sciatic nerve. The initial conservative treatment of injections in the conflict zone and the intervertebral joints was insufficient. Surgical exploration was undertaken via a posterior approach, demonstrating the conflict between the nerve and the ossified ligament. Resection of the ossified ligament eliminated the symptoms at the cost of transitory buttock hypesthesia. The anatomic and pathologic exam identified simple osseous metaplasia. Lumbar-pelvic coxometry analysis showed sagittalization of the acetabular roofs, excessive bilateral femoral torsion, and a step-off attenuation at the anterior cervicocephalic junction (low anterior cervico-cephalic femoral offset). In addition, reduced lumbar lordosis and accentuated sacral slope were noted, factors that could be related to modifications in the static balance of the lumbar-pelvic complex. This imbalance could be correlated to an increase in the forces applied to the pelvic ligament layers. The etiological hypothesis retained was that this osseous metaplasia was a reaction to excessive stresses on the sacrospinous ligament.

Climate change, flow regulation and land-use effects on the hydrology of the Peace-Athabasca-Slave System; findings from the Northern Rivers Ecosystem Initiative.

The Northern Rivers Ecosystem Initiative (NREI) was established in the late 1990s to address important science questions resulting from previous studies undertaken by the Northern Rivers Basin Study (NRBS). This manuscript summarizes the results from a number of reports on hydrologic research conducted on the Peace-Athabasca-Slave river and lake systems. Specific concerns expressed by the NRBS and subsequent NREI focused on how these systems were being affected by climate change, flow regulation and land-use changes. Issues addressed in this report include: the fate of aquatic perched basins within the Peace-Athabasca Delta under historical and future climate trends; the sources of major floods that replenish these basins and how the frequency, magnitude and source areas of such events have changed over time; the synoptic weather patterns and atmospheric teleconnections that are responsible for the generation of major snowmelt runoff that drive major floods; the potential effect that climate and land-use changes might have on basin runoff and delta lake levels; the specific hydro-climatic conditions required to produce major ice-jam floods on the Peace River and how these may be altered by climate change; remote-sensing methods to document delta flooding and vegetation change; and the dual effect of climate and flow regulation on the water levels of Great Slave Lake and how these may affect other nearshore processes, particularly wind seiches, that influence flooding of the Slave River Delta. A review of the major findings and recommendations for future research concludes the report.