Extracellular Vesicles Secreted by Pre-Hatching Bovine Embryos Produced In Vitro and In Vivo Alter the Expression of IFNtau-Stimulated Genes in Bovine Endometrial Cells.

The embryo-maternal interaction occurs during the early stages of embryo development and is essential for the implantation and full-term development of the embryo. In bovines, the secretion of interferon Tau (IFNT) during elongation is the main signal for pregnancy recognition, but its expression starts around the blastocyst stage. Embryos release extracellular vesicles (EVs) as an alternative mechanism of embryo-maternal communication. The aim of the study was to determine whether EVs secreted by bovine embryos during blastulation (D5-D7) could induce transcriptomic modifications, activating IFNT signaling in endometrial cells. Additionally, it aims to assess whether the EVs secreted by embryos produced in vivo (EVs-IVV) or in vitro (EVs-IVP) have different effects on the transcriptomic profiles of the endometrial cells. In vitro- and in vivo-produced bovine morulae were selected and individually cultured for 48 h to collect embryonic EVs (E-EVs) secreted during blastulation. E-EVs stained with PKH67 were added to in vitro-cultured bovine endometrial cells to assess EV internalization. The effect of EVs on the transcriptomic profile of endometrial cells was determined by RNA sequencing. EVs from both types of embryos induced several classical and non-classical IFNT-stimulated genes (ISGs) and other pathways related to endometrial function in epithelial endometrial cells. Higher numbers of differentially expressed genes (3552) were induced by EVs released by IVP embryos compared to EVs from IVV (1838). Gene ontology analysis showed that EVs-IVP/IVV induced the upregulation of the extracellular exosome pathway, the cellular response to stimulus, and the protein modification processes. This work provides evidence regarding the effect of embryo origin (in vivo or in vitro) on the early embryo-maternal interaction mediated by extracellular vesicles.

Semen parameter variability among users of at-home sperm testing kits.

BACKGROUND: Despite the generally accepted World Health Organization guidelines on semen analysis, an individual's results can display significant variation when performed across time or in different laboratories. Semen parameters are in fact highly variable measures that can differ significantly between various analyses. Numerous researchers have discovered a wide range of semen parameters within each individual male, but only a few studies included the analysis of semen parameters variability in patients with infertility. The aim of this study was to evaluate the inter- and intra-individual variability of semen parameters in men of reproductive age with normozoospermia and those with oligozoospermia. METHODS: Five hundred and thirteen who provided ≥ 2 semen samples (798 samples in total) using an at-home mail-in kit over a period of about 2 years were enrolled in the study. Semen samples collection using Give Legacy at-home mail-in semen collection kit; semen analysis at a CLIA-certified laboratory. RESULTS: The degree of intra-subject variation across all semen parameters was lower in men with normozoospermia compared to men with oligozoospermia. Men with normozoospermia furthermore demonstrated a level of intra-subject variation that was lower than inter-subject variation across all measured parameters. No association was observed between intra-subject coefficients of variation in any of the semen parameters, including sperm concentration, sperm count, motile sperm count, total motility, progressive motility, the percentage of sperm with normal morphology, and the age, duration of abstinence, and BMI of the men. CONCLUSION: The results of this observational study confirm the significant variability in semen parameters in men with normozoospermia and oligozoospermia, as measured from at-home semen collection kit samples. This further underscore the importance of securing multiple samples for analysis to provide a robust assessment of male fertility.

Lifestyle mediates the relationship between self-esteem and health-related quality of life in Chilean schoolchildren.

A healthy lifestyle, including food habits, physical activity (PA) and screen time (ST), is an important factor for well-being. The main purpose of this study was to analyze the association between lifestyle (i.e., PA, ST and food habits), self-esteem and health-related quality of life (HRQoL). A second objective was to determine the association between self-esteem with HRQoL, considering the mediating effect of lifestyle. A descriptive and cross-sectional study was performed, involving both girls (n = 282, 11.86 ± 0.82 years) and boys (n = 352, 12.02 ± 0.87 years). Lifestyle, self-esteem, HRQoL and anthropometrics parameters were evaluated. The study reported that self-esteem (ß; 0.04, P = 0.49) and PA (ß; 1.15, P < 0.001) had positive association with HRQoL. By contrast,ST was linked in an inverse way to HRQoL (ß; -1.82,, P < 0.001). According to the second objective, self-esteem had a significant association with HRQoL (total effect = 0.48, p < 0.01), and ST mediated this association negatively, instead, PA and food habits positively mediated this association. In conclusion, self-esteem presented association with HRQoL and lifestyle mediates this relationship positively (PA, MD adherence) and negatively (ST). Therefore, promoting healthy lifestyle among children should be a target of community- and school-based interventions to promote well-being.

Depression is associated with lower levels of physical activity, body image dissatisfaction, and obesity in Chilean preadolescents.

Mental health during the preadolescent years may be determinant for later periods of life; therefore, the factors that influence it need to be studied deeply. The aim of this study was to determine the association between depressive symptoms with physical activity (PA), body image dissatisfaction and weight status in preadolescents. A total of 269 girls and 329 boys (age 12.02 ± 0.98 years) were included in this study. Body mass index (BMI), waist circumference, body image dissatisfaction, PA and depressive symptoms were assessed. The PA reported an inverse association with depression in girls (ß:-2.12, P < 0.0001) and boys (ß:-1.13, P = 0.001). Instead, BMI in girls (ß: 1.00, P < 0.0001) and boys (ß: 0.70, P < 0.0001), and body image dissatisfaction in girls (ß: 0.05, P = 0.017) and boys (ß: 0.04, P = 0.003) reported positive association with depression. In the same way, depression was associated with obesity (odds ratio [OR] = 2.05, P = 0.020), but, the boys had higher risk (OR = 18.4, P < 0.001) than girls. In conclusion, depression in Chilean preadolescents presented association with low levels of PA, obesity and body image dissatisfaction. Therefore, schools should target and promote PA to improve psychological and physical health in preadolescents, which may reduce the future burden of mental illness.

PLCζ is the physiological trigger of the Ca2+ oscillations that induce embryogenesis in mammals but conception can occur in its absence.

Activation of the egg by the sperm is the first, vital stage of embryogenesis. The sperm protein PLCζ has been proposed as the physiological agent that triggers the Ca2+ oscillations that normally initiate embryogenesis. Consistent with this, recombinant PLCζ induces Ca2+ oscillations in eggs and debilitating mutations in the PLCZ1 gene are associated with infertility in men. However, there has been no evidence that knockout of the gene encoding PLCζ abolishes the ability of sperm to induce Ca2+ oscillations in eggs. Here, we show that sperm derived from Plcz1-/- male mice fail to trigger Ca2+ oscillations in eggs, cause polyspermy and thus demonstrate that PLCζ is the physiological trigger of these Ca2+ oscillations. Remarkably, some eggs fertilized by PLCζ-null sperm can develop, albeit at greatly reduced efficiency, and after a significant time-delay. In addition, Plcz1-/- males are subfertile but not sterile, suggesting that in the absence of PLCζ, spontaneous egg activation can eventually occur via an alternative route. This is the first demonstration that in vivo fertilization without the normal physiological trigger of egg activation can result in offspring. PLCζ-null sperm now make it possible to resolve long-standing questions in fertilization biology, and to test the efficacy and safety of procedures used to treat human infertility.

Capacitation increases glucose consumption in murine sperm.

Mammalian sperm acquire fertilization capacity in the female reproductive tract in a process known as capacitation. During capacitation, sperm change their motility pattern (i.e., hyperactivation) and become competent to undergo the acrosome reaction. We have recently shown that, in the mouse, sperm capacitation is associated with increased uptake of fluorescently labeled deoxyglucose and with extracellular acidification suggesting enhanced glycolysis. Consistently, in the present work we showed that glucose consumption is enhanced in media that support mouse sperm capacitation suggesting upregulation of glucose metabolic pathways. The increase in glucose consumption was modulated by bicarbonate and blocked by protein kinase A and soluble adenylyl cyclase inhibitors. Moreover, permeable cyclic adenosine monophosphate (cAMP) agonists increase glucose consumption in sperm incubated in conditions that do not support capacitation. Also, the increase in glucose consumption was reduced when sperm were incubated in low calcium conditions. Interestingly, this reduction was not overcome with cAMP agonists. Despite these findings, glucose consumption of sperm from Catsper1 knockout mice was similar to the one from wild type suggesting that other sources of calcium are also relevant. Altogether, these results suggest that cAMP and calcium pathways are involved in the regulation of glycolytic energy pathways during murine sperm capacitation.

The tyrosine kinase FER is responsible for the capacitation-associated increase in tyrosine phosphorylation in murine sperm.

Sperm capacitation is required for fertilization. At the molecular level, this process is associated with fast activation of protein kinase A. Downstream of this event, capacitating conditions lead to an increase in tyrosine phosphorylation. The identity of the tyrosine kinase(s) mediating this process has not been conclusively demonstrated. Recent experiments using stallion and human sperm have suggested a role for PYK2 based on the use of small molecule inhibitors directed against this kinase. However, crucially, loss-of-function experiments have not been reported. Here, we used both pharmacological inhibitors and genetically modified mice models to investigate the identity of the tyrosine kinase(s) mediating the increase in tyrosine phosphorylation in mouse sperm. Similar to stallion and human, PF431396 blocks the capacitation-associated increase in tyrosine phosphorylation. Yet, sperm from Pyk2(-/-) mice displayed a normal increase in tyrosine phosphorylation, implying that PYK2 is not responsible for this phosphorylation process. Here, we show that PF431396 can also inhibit FER, a tyrosine kinase known to be present in sperm. Sperm from mice targeted with a kinase-inactivating mutation in Fer failed to undergo capacitation-associated increases in tyrosine phosphorylation. Although these mice are fertile, their sperm displayed a reduced ability to fertilize metaphase II-arrested eggs in vitro.

Preventing metabolic syndrome in morbid obesity with resistance training: Reporting interindividual variability.

BACKGROUND AND AIMS: Resistant training (RT) improves health markers in obesity, but its effects in morbid obesity are unknown. We aimed to determine the effects of a RT-program in preventing/attenuating the metabolic syndrome (MetS) in patients with morbid obesity. A second aim was to report the interindividual variability in terms of improvements in MetS markers and other related co-variables. METHODS AND RESULTS: Twenty-one adults with obesity or morbid obesity were divided into two groups based on body mass index (BMI): a control obesity (CO, n = 7, BMI ≥35 < 40.0 kg/m2) and a morbid obese group (MO, n = 14, BMI ≥40 kg/m2). Participants completed a 20-week RT-program (3 sessions/week, 4-8 exercise) using free weights. Participants were assessed for MetS markers (waist circumference, systolic and diastolic blood pressure [BP], fasting glucose, high-density lipoproteins, and triglycerides) and other co-variables (total cholesterol, low-density lipoprotein, one-maximum repetition of biceps curl, and handgrip strength, 6 min walking test). Significant reductions in MetS markers were observed in both CO and MO groups (P < 0.05 to P < 0.0001), but significant reductions in diastolic BP and increases in HDL-C were noted only in the MO group (P < 0.0001). Changes in waist circumference, and systolic and diastolic BP were significantly greater only in the MO group (P < 0.001), but the CO group presented a greater fasting glucose decreases (P < 0.0001). The prevalence of non-responders between CO and MO groups was similar in the MetS outcomes. CONCLUSIONS: RT promotes greater improvements in overall MetS outcomes waist circumference, BP, and plasma triglycerides in patients with morbid obesity than in obese peers, with no overall differences in the prevalence of non-responders. CLINICAL TRIAL NUMBER: NCT03921853 at www.clinicaltrials.gov.

Tissue plasminogen activator (tPA) of paternal origin is necessary for the success of in vitro but not of in vivo fertilisation in the mouse.

Besides its fibrinolytic function, the plasminogen-plasmin (PLG-PLA) system is also involved in fertilisation, where plasminogen activators bind to plasminogen to produce plasmin, which modulates sperm binding to the zona pellucida. However, controversy exists, depending on the species, concerning the role of the different components of the system. This study focused its attention on the role of the PLG-PLA system on fertilisation in the mouse with special attention to tissue plasminogen activator (tPA). The presence of exogenous plasminogen reduced invitro fertilisation (IVF) rates and this decline was attenuated by the presence of plasmin inhibitors in combination with plasminogen. The incubation of spermatozoa with either oocytes or cumulus cells together with plasminogen did not change the acrosome reaction but reduced the number of spermatozoa attached. When spermatozoa from tPA-/- mice were used, the IVF rate decreased drastically, although the addition of exogenous tPA during gamete co-incubation under invitro conditions increased fertilisation success. Moreover, fertility could not be restored after invivo insemination of tPA-/- spermatozoa in the female ampulla, although tPA-/- males were able to fertilise invivo. This study suggests a regulatory role of the PLG-PLA system during fertilisation in the mouse with possible implications in human reproduction clinics, such as failures in tPA production, which could be partially resolved by the addition of exogenous tPA during IVF treatment.

The influence of cardiometabolic risk factors on cardiorespiratory fitness in volunteer Chilean firefighters.

OBJECTIVES: Maximum oxygen consumption (VO2max ) plays a fundamental role in firefighters' occupational activities due to the high intensity tasks they perform in their professional duties. In Chile, firefighters are volunteers (non-salary) and their lack of continuous and programmed physical activity may affect their physical fitness and health. The goal of this study was to determine the influence of anthropometric parameters and cardiometabolic risk (CMR) factors on the cardiorespiratory fitness (CRF) of volunteer Chilean firefighters. METHODS: Seventy-six volunteer male firefighters (median [5-95 percentiles]) aged 27.5 years [26-56], body mass index (BMI) 27.7 kg m-2 [19.9-35], and VO2max 44 mL kg-1 min-1 [36-56]) participated in the study. The following variables were assessed: BMI, fat mass%, body density, waist circumference (WC), waist-to-hip ratio (WHR), blood pressure, blood glucose, and VO2max . RESULTS: In total, 68% of the samples were overweight or obese. A total of 36% presented abdominal obesity (WC ≥102 cm). High blood pressure (HBP) was observed in 25% of firefighters and high blood glucose was found in about 20%. The presence of abdominal obesity was the strongest predictor of VO2max (OR = 12.35, 95% CI = 3.56-42.82, P < .001), followed by the WHR (OR = 11.5, 95% CI = 3.1-42.7, P < .001) and high blood glucose (OR = 2.87, 95% CI = 1.7-7.3, P = 0.019). CONCLUSION: This study showed that abdominal obesity in firefighters was the strongest predictor of low CRF. In addition, CRF was associated with CMR factors, except for HBP.

Feasibility of incorporating high-intensity interval training into physical education programs to improve body composition and cardiorespiratory capacity of overweight and obese children: A systematic review.

BACKGROUND/OBJECTIVE: High-intensity interval training (HIIT) can produce similar or improved results compare with traditional training, but the question as to whether HIIT can be used in the setting of physical education (PE) remains unanswered. The aim of this systematic review was to critically analyze the feasibility of incorporating HIIT programs into PE classes to improve the body compositions and cardiorespiratory fitness of overweight students. METHODS: We conducted database searches for literature dating between January 2012 and January 2017. Of the final six studies selected, three were conducted in children under 12 years old and three involved adolescents between 12 and 18 years old. RESULTS: The HIIT protocols consisted of 2-3 sessions per week, with intervals of 15 s and passive or active rests of 15 s, totaling up to 6 min of work with 4 min of rest. The duration of HIIT programs was 6-24 weeks. Significant changes were reported in body composition, body mass index, body fat (%), waist circumference, and sum of skinfolds; and increases in muscle mass were observed. The inclusion of HIIT programmes improved maximal oxygen uptake (VO2max), performance in the intermittent Yo-Yo test and maximal aerobic speed. CONCLUSIONS: The HIIT programmes showed improvements in the variables studied, with interventions two or three times weekly. Therefore, they can be used in schools, as a strategy to combat the childhood obesity pandemic and HIIT can be use alongside with existing PE activities within the same lesson or in specific periods during day school.

Discovery of LRE1 as a specific and allosteric inhibitor of soluble adenylyl cyclase.

The prototypical second messenger cAMP regulates a wide variety of physiological processes. It can simultaneously mediate diverse functions by acting locally in independently regulated microdomains. In mammalian cells, two types of adenylyl cyclase generate cAMP: G-protein-regulated transmembrane adenylyl cyclases and bicarbonate-, calcium- and ATP-regulated soluble adenylyl cyclase (sAC). Because each type of cyclase regulates distinct microdomains, methods to distinguish between them are needed to understand cAMP signaling. We developed a mass-spectrometry-based adenylyl cyclase assay, which we used to identify a new sAC-specific inhibitor, LRE1. LRE1 bound to the bicarbonate activator binding site and inhibited sAC via a unique allosteric mechanism. LRE1 prevented sAC-dependent processes in cellular and physiological systems, and it will facilitate exploration of the therapeutic potential of sAC inhibition.

Defective sperm head decondensation undermines the success of ICSI in the bovine.

The efficiency of intracytoplasmic sperm injection (ICSI) in the bovine is low compared to other species. It is unknown whether defective oocyte activation and/or sperm head decondensation limit the success of this technique in this species. To elucidate where the main obstacle lies, we used homologous and heterologous ICSI and parthenogenetic activation procedures. We also evaluated whether in vitro maturation negatively impacted the early stages of activation after ICSI. Here we showed that injected bovine sperm are resistant to nuclear decondensation by bovine oocytes and this is only partly overcome by exogenous activation. Remarkably, when we used heterologous ICSI, in vivo-matured mouse eggs were capable of mounting calcium oscillations and displaying normal PN formation following injection of bovine sperm, although in vitro-matured mouse oocytes were unable to do so. Together, our data demonstrate that bovine sperm are especially resistant to nuclear decondensation by in vitro-matured oocytes and this deficiency cannot be simply overcome by exogenous activation protocols, even by inducing physiological calcium oscillations. Therefore, the inability of a suboptimal ooplasmic environment to induce sperm head decondensation limits the success of ICSI in the bovine. Studies aimed to improve the cytoplasmic milieu of in vitro-matured oocytes and to replicate the molecular changes associated with in vivo capacitation and acrosome reaction will deepen our understanding of the mechanism of fertilization and improve the success of ICSI in this species.

Adaptaciones al ejercicio físico en el perfil lipídico y la salud cardiovascular de obesos mórbidos.

OBJECTIVE: To assess the effects of a physical exercise program on the lipid profile, weight status and the cardiovascular health of obese candidates for bariatric surgery. METHOD: 22 morbidly obese patients participated in a 6-month physical exercise program. 16 formed the adherent group (attendance ≥ 80%; age: 37.81 ± 11.90 years) and six the non-adherent group (attendance < 80%; age: 45.83 ± 14.23 years). Before and 72 h after the last session were evaluated: weight, body mass index (BMI), contour waist, blood pressure, fasting, cardio respiratory fitness, cholesterol and triglycerides. RESULTS: Weight, BMI and WC showed significant changes (p < 0.05) in both groups, with the results of the adherent group being better. In the adherent group, cardiorespiratory capacity (p = 0.001) was also increased, while diastolic pressure (p = 0.011), basal glycemia (p = 0.021) and triglycerides decreased significantly (p < 0.001). The non-adherent group did not present significant changes in these variables (p ≥ 0.05). CONCLUSIONS: The intervention was feasible to perform without negative effects for the participants, and it is possible to recommend improving the cardiovascular health of these of patients.

Biphasic role of calcium in mouse sperm capacitation signaling pathways.

Mammalian sperm acquire fertilizing ability in the female tract in a process known as capacitation. At the molecular level, capacitation is associated with up-regulation of a cAMP-dependent pathway, changes in intracellular pH, intracellular Ca(2+), and an increase in tyrosine phosphorylation. How these signaling systems interact during capacitation is not well understood. Results presented in this study indicate that Ca(2+) ions have a biphasic role in the regulation of cAMP-dependent signaling. Media without added Ca(2+) salts (nominal zero Ca(2+)) still contain micromolar concentrations of this ion. Sperm incubated in this medium did not undergo PKA activation or the increase in tyrosine phosphorylation suggesting that these phosphorylation pathways require Ca(2+). However, chelation of the extracellular Ca(2+) traces by EGTA induced both cAMP-dependent phosphorylation and the increase in tyrosine phosphorylation. The EGTA effect in nominal zero Ca(2+) media was mimicked by two calmodulin antagonists, W7 and calmidazolium, and by the calcineurin inhibitor cyclosporine A. These results suggest that Ca(2+) ions regulate sperm cAMP and tyrosine phosphorylation pathways in a biphasic manner and that some of its effects are mediated by calmodulin. Interestingly, contrary to wild-type mouse sperm, sperm from CatSper1 KO mice underwent PKA activation and an increase in tyrosine phosphorylation upon incubation in nominal zero Ca(2+) media. Therefore, sperm lacking Catsper Ca(2+) channels behave as wild-type sperm incubated in the presence of EGTA. This latter result suggests that Catsper transports the Ca(2+) involved in the regulation of cAMP-dependent and tyrosine phosphorylation pathways required for sperm capacitation.

Flow cytometry analysis reveals that only a subpopulation of mouse sperm undergoes hyperpolarization during capacitation.

To gain fertilizing capacity, mammalian sperm should reside in the female tract for a period of time. The physiological changes that render the sperm able to fertilize are known as capacitation. Capacitation is associated with an increase in intracellular pH, an increase in intracellular calcium, and phosphorylation of different proteins. This process is also accompanied by the hyperpolarization of the sperm plasma membrane potential (Em). In the present work, we used flow cytometry to analyze changes in sperm Em during capacitation in individual cells. Our results indicate that a subpopulation of hyperpolarized mouse sperm can be clearly distinguished by sperm flow cytometry analysis. Using sperm bearing green fluorescent protein in their acrosomes, we found that this hyperpolarized subpopulation is composed of sperm with intact acrosomes. In addition, we show that the capacitation-associated hyperpolarization is blocked by high extracellular K(+), by PKA inhibitors, and by SLO3 inhibitors in CD1 mouse sperm, and undetectable in Slo3 knockout mouse sperm. On the other hand, in sperm incubated in conditions that do not support capacitation, sperm membrane hyperpolarization can be induced by amiloride, high extracellular NaHCO3, and cAMP agonists. Altogether, our observations are consistent with a model in which sperm Em hyperpolarization is downstream of a cAMP-dependent pathway and is mediated by the activation of SLO3 K(+) channels.

Compartmentalization of distinct cAMP signaling pathways in mammalian sperm.

Fertilization competence is acquired in the female tract in a process known as capacitation. Capacitation is needed for the activation of motility (e.g. hyperactivation) and to prepare the sperm for an exocytotic process known as acrosome reaction. Although the HCO3(-)-dependent soluble adenylyl cyclase Adcy10 plays a role in motility, less is known about the source of cAMP in the sperm head. Transmembrane adenylyl cyclases (tmACs) are another possible source of cAMP. These enzymes are regulated by stimulatory heterotrimeric Gs proteins; however, the presence of Gs or tmACs in mammalian sperm has been controversial. In this study, we used Western blotting and cholera toxin-dependent ADP-ribosylation to show the Gs presence in the sperm head. Also, we showed that forskolin, a tmAC-specific activator, induces cAMP accumulation in sperm from both WT and Adcy10-null mice. This increase is blocked by the tmAC inhibitor SQ22536 but not by the Adcy10 inhibitor KH7. Although Gs immunoreactivity and tmAC activity are detected in the sperm head, PKA is only found in the tail, where Adcy10 was previously shown to reside. Consistent with an acrosomal localization, Gs reactivity is lost in acrosome-reacted sperm, and forskolin is able to increase intracellular Ca(2+) and induce the acrosome reaction. Altogether, these data suggest that cAMP pathways are compartmentalized in sperm, with Gs and tmAC in the head and Adcy10 and PKA in the flagellum.

Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation.

Mammalian sperm require time in the female tract in order to be able to fertilize an egg. The physiological changes that render the sperm able to fertilize are known as capacitation. Capacitation is associated with an increase in intracellular pH, an increase in intracellular calcium and phosphorylation of different proteins. This process is also accompanied by the hyperpolarization of the sperm plasma membrane potential. Recently, we presented evidence showing that epithelial Na+ channels (ENaC) are present in mature sperm and that ENaCs are blocked during capacitation. In the present work, we used flow cytometry to analyze changes in intracellular Na+ concentration ([Na+](i)) during capacitation in individual cells. Our results indicate that capacitated sperm have lower Na+ concentrations. Using sperm with green fluorescent protein in their acrosomes, it was shown that the lower [Na+](i) concentration only occurs in sperm having intact acrosomes. ENaC inhibition has been shown in other cell types to depend on the activation of cystic fibrosis transmembrane conductance regulator (CFTR). In non-capacitated sperm, amiloride, an ENaC inhibitor, and genistein, a CFTR activator, caused a decrease in [Na+](i), suggesting that also in these cells [Na+](i) is dependent on the crosstalk between ENaC and CFTR. In addition, PKA inhibition blocked [Na+](i) decrease in capacitated sperm. Altogether, these data are consistent with the hypothesis that the capacitation-associated hyperpolarization involves a decrease in [Na+](i) mediated by inhibition of ENaC and regulated by PKA through activation of CFTR channels.

Unveiling the Role of IGF-I in Fertility: Effect of Long-Acting Bovine Somatotropin (bST) on Terminal Follicular Development and Fertility during an Annual Reproductive Cycle in Sheep.

The study aimed to assess the effect of long-acting bST treatment, in a dose that only increases IGF-I plasma concentrations, on ovarian and fertility markers of estrous synchronized ewes that were fed to keep their bodyweight. Three experiments were designed to evaluate this effect: in Experiment 1, 18 ewes were distributed in groups (bST 0, 30, 50 mg) to measure plasma IGF-I and insulin for 15 days; in Experiment 2, 92 ewes (5 replicates) in two groups (0 and 30 mg bST) were synchronized using a 6-day progesterone protocol during the breeding season to assess the effect of bST on follicular and luteal performances, estrous and ovulation, and fertility after mating. In Experiment 3, 50 ewes (3 replicates) were used to repeat the study before but during anestrus. Results indicate that 50 mg bST increased IGF-I and insulin plasma concentrations, but 30 mg bST only increased IGF-I concentrations; and that only during the breeding season did 30 mg bST increase the number of lambs born and the reproductive success of ovulatory-sized follicles compared to controls. This occurred without it affecting any other reproductive marker. In conclusion, 30 mg bST treatment may improve oocyte competence for fertility during the breeding season.

A Subovulatory Dose of Human Chorionic Gonadotropin (hCG) May Sustain Terminal Follicle Development and Reproductive Efficiency during Anestrus in Sheep.

The study tested the hypothesis that a single administration of hCG supports the LH-dependent phase of terminal follicular development in synchronized sheep during anestrus, using eCG as a functional reference. Using a clinical approach, four experiments were designed to achieve the following: (1) Identify the inhibitory influence of anestrus on reproduction efficiency; (2) Assess the potential of hCG to keep functional blood concentrations after a single dose; (3) Characterize the effect of different doses of hCG on reproductive functional markers; (4) To compare the ability of hCG to that of eCG to support follicular development and fertility based on the same markers. The results showed that anestrus seems to affect follicular and luteal function under LH dependency as FSH-dependent markers are not compromised; hCG maintains higher blood concentrations than controls for at least 48 h; hCG improves follicular development and ovulatory rates compared to controls and at standards comparable to a breeding season; and ewes treated with hCG exhibit similar performance to those treated with eCG. Our results conclude that hCG can be used to support follicular function during anestrus in sheep, aiming to perfect its regulation in assisted reproduction.