Is sperm DNA fragmentation (SDF) involved in the lower fertilising ability of cryopreserved spermatozoa from cancer patients? In this study we evaluated SDF in two different cytometric sperm populations (PIbrighter and PIdimmer). In cryopreserved spermatozoa from cancer patients, PIbrighter SDF, which is more relevant to reproduction, was positively related to sperm motility and that the proportion of these cells was higher in samples from cancer versus non-cancer (oligozoospermic or affected by autoimmune pathologies) patients. The results of the study indicate that thawed motile spermatozoa of cancer patients may carry DNA damage.
Reproduction, Fertility and Development
Volume 29 Number 4 2017
RD15103Cytokine gene expression at the maternal–fetal interface after somatic cell nuclear transfer pregnancies in small ruminants
The reasons why somatic cell nuclear transfer (SCNT) pregnancies are at increased risk of pregnancy loss are poorly understood. This study investigated pregnancy losses and gene expression of inflammatory mediators in sheep and goat SCNT pregnancies. Sheep SCNT pregnancies had an increased incidence of pregnancy loss and increased expression of inflammatory cytokines in the placenta, which indicates the presence of an unfavourable environment for fetal development.
RD15182Follicular fluid and serum metabolites in Holstein cows are predictive of genetic merit for fertility
This study examined the metabolite profile of serum and follicular fluid on Day 7 of the oestrous cycle in Holstein cows with good or poor genetic merit for fertility. Differences between the two genotypes in the abundance of fatty acids and amino acids were highly predictive of fertility. The results demonstrate that metabolite profiles in cattle respond to genetic selection for fertility traits, and specific metabolites may be useful biomarkers for fertility
RD15157Is the association between insulin resistance and diabetogenic haematopoietically expressed homeobox (HHEX) polymorphism (rs1111875) affected by polycystic ovary syndrome status?
Insulin resistance (IR) is an underlying factor, common to both polycystic ovary syndrome (PCOS) and type 2 diabetes. Genetic factors have also been reported to play a role in the pathogenesis of these two conditions, indicting a common genetic aetiology. In this study we aim to determine whether the genetic association between IR and diabetogenic SNPs is affected by PCOS status in Iranian women and found that the association may be influenced by the condition.
RD15159Cumulus cell-conditioned medium supports embryonic stem cell differentiation to germ cell-like cells
The effect of cumulus cell-conditioned medium (CCM) on the differentiation of buffalo embryonic stem cells to germ cells were explored. A concentration of 20%–40% conditioned medium induced the highest expression of primordial germ cell, meiotic, spermatocyte and oocyte markers and led to the formation of oocyte-like structures. Further development of the cumulus conditioned medium induced differentiation strategy would be useful for understanding mammalian gametogenesis and transgenic animal production.
Infertility affects 1 in 10 couples or millions of people worldwide and while IVF is an important intervention, 75% of all embryos transferred into the uterus do not establish a pregnancy. We investigated how the uterus of infertile women may prevent pregnancy and discovered a protein released by uterine tissue that potentially blocks pregnancy. Targeting this protein may be useful as a new treatment option for infertile women.
The aquaporin family plays a fundamental role in water and solute transport across biological membranes. The present work has studied, for the first time, the presence and localisation of aquaglyceroporin 3 in boar spermatozoa. Although two different localisation patterns for this protein were identified, neither AQP3 content nor its localisation were found to be correlated with sperm quality parameters.
We studied for the first time the effect of high inbreeding levels in cattle sperm by automatised and objective methods. An extensive genetic analysis using an animal model showed that inbred bulls have a reduced field fertility and an increased calving interval compared with non-inbred bulls. Sperm from high inbred animals showed an increased ‘hyper-active like’ movement that could explain their decreased fertility.
RD15350In vitro maturation affects chromosome segregation, spindle morphology and acetylation of lysine 16 on histone H4 in horse oocytes
Mammalian eggs are often affected by aneuploidy, leading to failure in embryo implantation, miscarriages and genetic diseases. In horse oocytes the onset of aneuploidy is correlated with the disruption of an epigenetic mark that occurs without changes in gene expression. A deeper knowledge of the molecular basis of aneuploidy will allow the development of safer treatments to obtain healthy eggs.
To determine whether body weight has an effect on semen quality, the present study explored the possible association between body weight (in terms of body mass index) and sperm function. The results indicate that both morbid obesity and being underweight have a negative effect on sperm quality, probably as a result of changes to epididymal function. These findings highlight the importance of an adequate body weight as the best option for fertility.
RD15349Elimination of methylation marks at lysines 4 and 9 of histone 3 (H3K4 and H3K9) of spermatozoa alters offspring phenotype
The function of epigenetic marks present in retained histones of mature spermatozoa is largely unknown. By reducing the methylation level at lysines 4 and 9 of histone 3 of mature spermatozoa we aimed to shed light on their significance during development. We discovered that these marks are important for normal embryo development and their reduction leads to abortions, premature death and morphological abnormalities in the offspring.
RD15334Effects of prepartum oilseed supplements on subclinical endometritis, pro- and anti-inflammatory cytokine transcripts in endometrial cells and postpartum ovarian function in dairy cows
Dietary fats are known to influence immune cell function and fertility. The effects of pre-calving diets containing oilseeds (sunflower or canola vs. no oilseed) on uterine inflammation and ovarian function in dairy cows were investigated. Adding sunflower seed to pre-calving diets positively influenced ovarian function without affecting uterine inflammation, suggesting a role for omega-6 fatty acids in nutritional management of reproduction.
The average age for a primipara has been increased and maternal age affects the quality of oocytes. The present study addressed whether age-associated deterioration of oocytes stems from the oocyte itself or the surrounding environment and found that follicular fluid plays a major role. On the basis of the results, given optimum conditions, oocytes derived from aged females could give rise to high-quality oocytes.
RD15230Mammalian target of rapamycin/eukaryotic initiation factor 4F pathway regulates follicle growth and development of theca cells in mice
Follicular growth is mediated by gonadotropic hormones, growth factors and signalling pathways, such as the mammalian target of rapamycin (mTOR) signalling pathway. However, the molecular mechanisms underlying the development of theca cells have rarely been studied. Herein we provide unbiased, detailed and convincing evidence (microarray, in vitro and in vivo studies) to demonstrate that mTOR/eukaryotic initiation factor 4F signalling plays an important role in follicular growth, particularly the development of thecal cells.
RD15243Active 3ʹ–5ʹ cyclic nucleotide phosphodiesterases are present in detergent-resistant membranes of mural granulosa cells
Lipids rafts are specialised membrane microdomains involved in cell signalling and phosphodiesterases are key enzymes involved in cyclic AMP signalling in the ovary. We found that in mural granulosa cell membranes, the cyclic AMP–phosphodiesterase activity is mainly contributed by the rafts. These results suggest that in mural granulosa cells, lipid rafts exist as a cell-signalling platform.
RD15406LIM kinase activity is required for microtubule organising centre positioning in mouse oocyte meiosis
In female meiosis, accurate chromosome separation is driven by the spindle apparatus, which is organised under the regulation of unique microtubule organising centres (MTOCs). LIM kinase 1 activity is required for the establishment and maintenance of MTOC bipolarity in mouse oocytes during meiotic division. The results of the present study provide an insight into the molecular mechanism governing bipolar spindle formation in oocytes during meiotic maturation.
RD15289Effect of spermatozoa motility hyperactivation factors and gamete coincubation duration on in vitro bovine embryo development using flow cytometrically sorted spermatozoa
This study evaluated the effects of sperm motility enhancers, media and different IVF times on embryo development. The sperm motility enhancers caffeine and theophylline improved cleavage and embryo development rates, synthetic oviducal fluid fertilisation medium outperformed M199 fertilisation medium and shortening the oocyte–sperm coincubation time (from 18 to 8 h) resulted in similar overall embryo performance rates. This study provides important information for optimising IVF protocols.
RD15480Obesity causes weight increases in prepubertal and pubertal male offspring and is related to changes in spermatogenesis and sperm production in rats
Obesity, a global problem, adversely affects human health including reproduction. This study investigated whether exposure to an obesogenic environment during prenatal and postnatal periods affects the initiation and progress of spermatogenesis in rats. It is evident that an obese condition decreases counts of spermatids and spermatozoa and serum levels of testosterone and increases the proportion of abnormal spermatozoa, which might reduce reproductive potential.
RD15367Growth arrest specific 1 (Gas1) and glial cell line-derived neurotrophic factor receptor α1 (Gfrα1), two mouse oocyte glycosylphosphatidylinositol-anchored proteins, are involved in fertilisation
In addition to oocyte Cd9 tetraspanin, Juno, a glycosylphosphatidylinositol (GIP)-anchored protein, has recently been discovered to be essential to mammalian fertilisation, as a sperm receptor for Izumo1. The present study demonstrates the involvement of another two other oocyte GPI-anchored proteins, namely growth arrest specific 1 (Gas1) and glial cell line-derived neurotrophic factor receptor α1 (Gfrα1), in fertilisation. The results reinforce the implication of several molecules forming probably complexes.
Monozygotic (MZ) twins are of great interest to elucidate the contributions of pre- and postnatal environmental factors on the expression of complex traits and diseases. In this study we investigated variations in sperm DNA methylation between MZ twin bulls to understand the observed differences in offspring productivity of MZ twin bulls despite their identical genetic backgrounds. Variation in sperm DNA methylation between MZ twin bulls was observed and may contribute to incongruous diverging performances of daughters sired by these bulls.
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RD16144Prostaglandin F2α–PTGFR signalling activation, growth factor expression and cell proliferation in bovine endometrial explants
The association between bovine endometrial growth and prostaglandin (PG) F2α–PGF2α receptor (PTGFR) signalling activation is unknown. In the present study, cell proliferation and the expression of growth factors essential for endometrial growth were upregulated by PGF2α-PTGFR signalling activation by fluprostenol in bovine endometrial explants in vitro. These results indicate that PGF2α-PTGFR signalling activation could be involved in endometrial growth.
RD16477Tissue-specific inhibition of urokinase-type plasminogen activator expression in the testes of mice by inducible lentiviral RNA interference causes male infertility
uPA shRNA lentivirus system induced by Dox inhibited the expression of uPA in TM4 cells. Through injecting the lentivirus into mice testis, we found that downregulation of uPA in mice testis decreased the fertility of male mice, which may be caused by a reduction in sperm motility. The inducible uPA shRNA system is safe and reversible to regulate male fertility.
RD16362Evaluation of SmartFlare probe applicability for verification of RNAs in early equine conceptuses, equine dermal fibroblast cells and trophoblastic vesicles
Live cell RNA imaging in mammalian embryos would represent a quantum leap in understanding cell differentiation and early embryonic development. Applicability of RNA imaging by SmartFlare technology was tested in equine embryos surrounded by an embryonic capsule and/or the zona pellucida, nude trophoblastic vesicles and fibroblast cell cultures. Encapsulated embryos block SmartFlare uptake, but stages surrounded by the zona pellucida only and nude vesicles incorporate the probes, leading to specific fluorescence suitable for tracing gene expression cascades necessary for further embryo development.
RD16426Effects of antifreeze glycoprotein 8 (AFGP8) supplementation during vitrification on the in vitro developmental capacity of expanded bovine blastocysts
Antifreeze glycoprotein 8 (AFGP8) supplementation during vitrification beneficially protects bovine blastocysts against chill-induced injury. AFGP8 supplementation during vitrification enhances the cryosurvival of bovine blastocysts. AFGP8 supplementation during vitrification improves the subsequent in vitro developmental capacity of bovine blastocysts.
RD16478Expression of protocadherin 11Yb (PCDH11Yb) in seminal germ cells is correlated with fertility status in men
Knowledge of molecular defects in spermatozoa becomes critical during assisted reproduction to treat male factor infertility. Herein we show that the absence or low expression of protocadherin 11Yb (PCDH11Yb) in spermatozoa is associated with male factor infertility. Profiling of a panel of differentially expressed sperm proteins, including PCDH11Yb, in men with infertility may be of diagnostic and therapeutic value in infertility management.
RD16383Localisation of stem cell factor, stanniocalcin-1, connective tissue growth factor and heparin-binding epidermal growth factor in the bovine uterus at the time of blastocyst formation
Limited information is available about the role of growth factors in very early embryonic development. This study demonstrates the presence of stem cell factor, stanniocalcin-1, connective tissue growth factor and heparin-binding epidermal growth factor-like growth factor in bovine uterine fluid, endometrium and embryos during blastocyst formation. The results suggest a role for these proteins in early embryo development in cattle.
RD16186Identification of differentially expressed placental transcripts during multiple gestations in the Eurasian beaver (Castor fiber L.)
It is still unspecified if and how the number of embryos or fetal sex affect pregnancy outcome. The present study determined whether there are differences in transcriptome profiles of the beaver placenta as a result of fetal sex or multiplicity of gestation. The results indicate that the number of fetuses affects the expression profile in the subplacental transcriptome. Enhancement of transcriptome resources will improve our understanding of the pathways relevant to proper placental development and successful reproduction in general.
RD16302Polydatin improves the developmental competence of bovine embryos in vitro via induction of sirtuin 1 (Sirt1)
Polydatin (PD) has antioxidant activity and is useful for embryo development in vitro. Supplementation of IVM medium with PD scavenged reactive oxygen species and improved embryo developmental competence via induction of sirtuin 1 (Sirt1).
RD16294Sperm gamma-aminobutyric acid type A receptor delta subunit (GABRD) and its interaction with purinergic P2X2 receptors in progesterone-induced acrosome reaction and male fertility
The mechanism underlying the non-genomic action of progesterone in sperm functions and related Ca2+ mobilisation remains elusive. Herein we report the expression of gamma-aminobutyric acid type A receptor delta subunit (GABRD) in human and rodent spermatozoa and its involvement in mediating the progesterone-induced acrosome reaction through its interaction with purinergic P2X2 receptors. GABRD represents a novel progesterone receptor or modulator in sperm responsible for the progesterone-induced Ca2+ influx required for the acrosome reaction.
RD16333Restoration of immune and renal function in aged females by re-establishment of active ovarian function
Immune and renal function, both critical for reproductive success, were evaluated to determine if exposure of postreproductive female mice to young ovaries would influence these non-reproductive physiological functions. Postreproductive mice received new (60-day-old) ovaries at 12 months of age and were evaluated at 16 months of age. Age-related declines in immune and renal function in 16-month-old control mice were restored to levels found in 6-month-old mice by ovarian transplantation. Re-establishment of reproductive function in aged female mice positively influenced non-reproductive functions.
RD16473In vitro addition of docosahexaenoic acid improves the quality of cooled but not frozen–thawed stallion semen
Improvements in the quality of stallion semen are required to increase the success of artificial insemination. The aim of this study was to assess the effects of the addition of docosahexaenoic acid to semen diluents on the quality of stallion semen. Docosahexaenoic acid increased spermatozoa motility and membrane fluidity in cooled semen and thus can be used to increase the quality of spermatozoa.
RD16327Gene-specific profiling of DNA methylation and mRNA expression in bovine oocytes derived from follicles of different size categories
The present study investigated DNA methylation profiles and mRNA expression of imprinted and non-imprinted candidate genes in bovine oocytes from antral follicles of three different size classes (≤2 mm, 3–5 mm, ≥6 mm) to unravel the epigenetic contribution to follicular and oocyte growth. We observed an increased number of aberrantly methylated alleles in bH19, bSNRPN and bDNMT3 Lo of oocytes from small antral follicles and an increased frequency of CpG sites with an unclear methylation status for DNMT3 Ls in large follicles. Results indicate that a follicle diameter of ~2 mm is critical for establishing DNA methylation profiles.
RD16193Microtubule stabilisers docetaxel and paclitaxel reduce spindle damage and maintain the developmental competence of in vitro-mature bovine oocytes during vitrification
The aim of this study was to compare the efficacy of paclitaxel (PT) and docetaxel (DT) in reducing spindle damage in bovine oocytes during vitrification. The important finding of the study was that pretreatment of bovine oocytes with 0.05 µM DT for 30 min before vitrification reduced spindle damage to a greater degree than pretreatment with 1.0 µM PT. This finding will be useful in improving oocyte cryopreservation in the future.
RD16233Extracellular-like matrices and leukaemia inhibitory factor for in vitro culture of human primordial follicles
The possibility of growing immature human ovarian follicles in laboratories will assist fertility restoration. The aim of the present study was to investigate whether growth of immature follicles can be promoted by using different culture surfaces and supplementation with a growth factor. It was unclear whether any of the culture surfaces had an advantage over the others. In addition, the growth factor did not promote growth. Further studies should test other culture surfaces and identify additional fluid supplements, so that an optimal system can be developed.
RD16283Poly(A)-binding proteins are required for translational regulation in vertebrate oocytes and early embryos
Poly(A)-binding proteins function in the timely regulation of gene expression during oocyte maturation, fertilisation and early embryo development in vertebrates. In this review, we comprehensively evaluate and discuss the expression patterns and particular functions of the embryonic poly(A)-binding protein (EPAB) and poly(A)-binding protein, cytoplasmic 1 (PABPC1) genes, especially in mouse and human oocytes and early embryos.
RD16276Epigenetic changes of hepatic glucocorticoid receptor in sheep male offspring undernourished in utero
Evidence exists that early life nutritional insults can predispose to metabolic disorders, like Type 2 diabetes, for which excessive hepatic glucose production is a contributing factor. Maternal undernutrition alters the epigenetic status of the glucocorticoid receptor and increases the expression of gluconeogenic enzymes in the liver of male offspring. Undernutrition in the womb programs long-lasting changes in specific gene expression that leads to persistent metabolic abnormalities.
RD16277Maternal obesity in the rat impairs male offspring aging of the testicular antioxidant defence system
Maternal obesity leads to premature aging of male offspring reproductive capacity and increases testes oxidative stress in male rat offspring. Our results show that maternal obesity modifies the antioxidant enzyme system in the testes of offspring, which, in turn, affects testicular physiological functions and leads to premature aging of male reproductive capacity.
Mitochondrial ferritin is an iron-storage protein highly expressed in human and mouse spermatozoa. This work showed that male mice, but not female, in which mitochondrial ferritin was genetically deleted have reduced fertility without significant alterations in sperm parameters. These data show that mitochondrial ferritin has a role in male fertility.
RD16198Ontogeny of clock and KiSS-1 metastasis-suppressor (Kiss1) gene expression in the prepubertal mouse hypothalamus
Kisspeptin neurons in the brain are crucial for the hormone surge required for ovulation, but are established before puberty. This study examined kisspeptin and brain clock gene rhythms (known to restrict kisspeptin action) in young mice. The data show that the kisspeptin peak is not seen prior to puberty and there is incomplete development of clock gene rhythmicity at this time.
RD16284Intrafollicular oestradiol production, expression of the LH receptor (LHR) gene and its isoforms, and early follicular deviation in Bos indicus
In this study we investigated the expression of the LH receptor (LHR) gene during dominant follicle selection in Bos indicus cattle. The main finding was that the early deviation of the dominant follicle in this subspecies is driven by the differential expression of full-length LHR and its isoforms in small follicles. This finding will contribute to explaining the differences in ovarian physiology between B. indicus and Bos taurus.
RD16087Gel-coated tubes extend above-freezing storage of honey bee (Apis mellifera) semen to 439 days with production of fertilised offspring
Honey bees, which are vital agricultural pollinators, are at risk from numerous factors. The ability to artificially inseminate these species provides a means to strengthen them genetically by collecting semen from bees of different regions throughout the world and then inseminating bees at distant locations. Our study aimed to substantially improve the viability of spermatozoa between collection and insemination.
RD15476Prepartum maternal diets supplemented with oilseeds alter the fatty acid profile in bovine neonatal plasma possibly through reduced placental expression of fatty acid transporter protein 4 and fatty acid translocase
Preferential transportation of essential fatty acids over non-essential fatty acids occurs from the mother to fetus in humans. However, little is known about placental transfer of fatty acids in the bovine. In the present study, prepartum dietary fat supplementation of cows reduced maternal long-chain polyunsaturated fatty acids, neonatal total omega-3 fatty acids, eicosapentaenoic and docosahexaenoic acids and total, fat possibly through reduced expression of certain placental fatty acid transporter genes.
RD16321Presence of vascular endothelial growth factor during the first half of IVM improves the meiotic and developmental competence of porcine oocytes from small follicles
Ovaries contain a large number of small follicles, but the meiotic and developmental competence of oocytes from these follicles is quite low. In the present study we demonstrated that the addition of 200 ng mL–1 vascular endothelial growth factor to the IVM medium increased the maturation rate of porcine oocytes from small follicles and that blastocyst formation following parthenogenetic activation also increased. These findings may contribute to efficient animal production and human assisted reproductive technology.
RD16181Modulation of adiponectin system expression in the porcine uterus during early pregnancy by prostaglandin E2 and F2?
Adiponectin is a hormone perceived as the link between energy metabolism and the reproductive system, but its interactions with factors responsible for regulation of uterine homeostasis during pregnancy remain unknown. Herein we demonstrated the effects of prostaglandins E2 and F2α on adiponectin and its receptors during early gestation. Recognition of these interactions will contribute to our understanding of the physiology of early pregnancy.
A diabetic pregnancy leads to distinct changes in maternal cholesterol metabolism, whereas embryonic cholesterol concentrations and cholesterol metabolism genes are not affected. The trophoblast : embryoblast cholesterol ratio suggests a higher metabolic activity of the embryoblast in diabetes. The findings of the present study imply independent and functional cholesterol metabolism in the mammalian embryo before implantation.
RD16382The current state of reproductive biology research in Australia and New Zealand: core themes from the Society for Reproductive Biology Annual Meeting, 2016
Reproductive science is central to our understanding of biology, translating to biological processes across many organ systems. The Society for Reproductive Biology conference review provides a focused summary of symposia at the 2016 annual meeting. The presentations highlight the breadth of reproductive research in Australia and New Zealand and its potential to affect the fertility and health of future generations.
RD16213Protein in culture and endogenous lipid interact with embryonic stages in vitro to alter calf birthweight after embryo vitrification and warming
Intracellular lipid is involved in long-term blastocyst competence to survive vitrification. We studied how protein removal affects bovine embryo development in vitro, lipid content and pregnancy outcomes, and found that calf weight at birth was altered depending on specific embryonic kinetics within vitrified–warmed embryos. Therefore, it is possible to modify calf phenotypes by introducing simple changes in culture conditions combined with selection of embryos.
RD16174Rosmarinic acid reverses the effects of metronidazole-induced infertility in male albino rats
Metronidazole, an essential medicine, adversely affects male fertility. We aimed at investigating the capability of Rosmarinic acid to reverse metranidazole induced male infertility. We found that certain doses of Rosmarinic acid could reverse metronidazole’s effects on sperm count, motility, morphology and testis ultrastructure. Our findings have great clinical implications where Rosmarinic acid might have a potential use in reversing Metronidazole induced male infertility.
RD15504Regulation of heat-inducible HSPA1A gene expression during maternal-to-embryo transition and in response to heat in in vitro-produced bovine embryos
Early mammalian embryos are sensitive to heat stress (HS) despite abundant oocytic iHSPA protein chaperones and mRNAs typically induced by stress. In this work, bovine HSPA1A gene transcription was detectable during embryo genome activation but induced by HS only after this stage, which coincides with better embryonic tolerance. Absence of stress-induced gene expression may therefore contribute to embryo sensitivity.
RD16297Effect of nutritionally induced hyperlipidaemia on in vitro bovine embryo quality depends on the type of major fatty acid in the diet
Hyperlipidemia associated with unbalanced diet and obesity is directly linked with reduced fertility. This study shows that substituting saturated fat with omega-3 fat in high caloric diets can alleviate detrimental effects of hyperlipidemic serum on bovine early embryo development and quality in vitro. This suggests that dietary manipulation of serum fatty acid profile may enhance fertility in patients with metabolic disorders.
The deleterious effects of heat stress on fertility are more pronounced in dairy cows than beef cows. The transcriptome profile in Nelore and Holstein oocytes subjected to heat shock during IVM and the mRNA abundance of selected genes in cumulus cells were determined. The analyses indicated that the deficiency in dairy cattle in terms of cellular protection against heat stress is regulated at the post-transcriptional level.
RD15535Sleep restriction in Wistar rats impairs epididymal postnatal development and sperm motility in association with oxidative stress
Sleep alterations are a public health problem worldwide that affect adolescents during puberty because of early school hours and interactive activities. In the present study, sleep restriction during peripuberty in rats affected epididymal postnatal development and sperm motility, in association with oxidative stress and a decrease in the epithelial compartment in the cauda epididymidis. The results of this study suggest a possibility for improving fertility after sleep restriction.
RD16223Effect and possible mechanisms of melatonin treatment on the quality and developmental potential of aged bovine oocytes
This research expands our understanding of the regulators that delay oocyte aging. Treatment of aged oocytes with exogenous melatonin improves oocyte quality, rearranges the cytoskeleton and improves developmental capacity. Melatonin is a good agent for delaying oocyte aging in vitro and preventing poor developmental competence of bovine oocytes in assisted reproductive technologies.
RD16183Induced sub-lethal oxidative damage affects osmotic tolerance and cryosurvival of spermatozoa
If the physiological balance between production and scavenging of reactive oxygen species (ROS) is shifted towards production of ROS this may result in accumulation of cell damage. The aim of this study was to establish whether sub-lethal oxidative damage negatively affects cryosurvival and pre-freeze sperm osmotic tolerance. Generation of defined levels of intracellular ROS can be used for testing strategies designed to counteract oxidative damage.
RD16310Reproductive dysfunction after mercury exposure at low levels: evidence for a role of glutathione peroxidase (GPx) 1 and GPx4 in male rats
Mercury has been identified as a risk factor for declines in sperm quality and infertility in humans. The present study delves further into the mechanisms of action of mercury, showing a role for glutathione peroxidase (GPx) 1 and GPx4 in the reproductive toxicity of mercury. The involvement of the glutathione system highlights this antioxidant system as a potential target of mercury after its accumulation in reproductive organs.
RD16037Ovariectomy increases the phenotypic plasticity of the female prostate epithelium in the Mongolian gerbil (Meriones unguiculatus)
After hormonal deprivation the female gerbil prostate undergoes several morphophysiological changes. This work identified and characterised the cellular heterogeneity of the female prostate epithelium in normal conditions and after ovariectomy. Secretory cells of the female prostate are not morphologically and functionally uniform, presenting a phenotypical plasticity according to the hormonal environment in which they operate.
RD16280Spatiotemporal expression profile of proteases and immunological, angiogenic, hormonal and apoptotic mediators in rat placenta before and during intrauterine trophoblast migration
The current literature does not provide a comprehensive profile of the factors involved in placental development in rats during intrauterine trophoblast migration. In this study we evaluated the gene and/or protein expression of several placental factors during the second half of pregnancy in rats and demonstrated that the expression of proteases and immunological, angiogenic, hormonal and apoptotic mediators is variable and depends on the stage of trophoblast migration. These changes in the expression profile are probably related to alterations in placental morphogenesis and function.
Infusion of the ovine uterus with hyaluronan (HA) during the pre-attachment period changes the expression of some adhesion markers at the embryo–maternal interface, resulting in reduced endometrial receptivity and failure of embryo attachment. In contrast, local inhibition of HA production in the uterus enhances embryo attachment.
RD15537Circulating anti-Müllerian hormone concentrations in relation to age and season in male and female beluga (Delphinapterus leucas)
Anti-Müllerian hormone (AMH) has been proposed as a biomarker of ovarian reserve in women and has been detected in many species; however, this is the first characterisation of AMH in a cetacean. AMH was described in males and females from the year of birth to over 30 years of age during different reproductive seasons, and a decrease in AMH in older animals (>30 years old) was detected. AMH may be of value for reproductive health monitoring of the species and as a biomarker of reproductive senescence in both sexes.
RD16200Non-surgical sterilisation methods may offer a sustainable solution to feral horse (Equus caballus) overpopulation
Culling feral horses is currently met with public criticism, which has created an urgent need to develop new methods of population control. Delivery of a contraceptive agent that causes sterility, without the need for surgery, is currently under investigation. These novel techniques may provide a sustainable and effective alternative for feral horse control.
RD16122A combination of insulin-like growth factor I (IGF-I) and FSH promotes proliferation of prepubertal bovine Sertoli cells isolated and cultured in vitro
Sertoli cells, which support and coordinate developing sperm in the testes, were isolated from 8-week-old Holstein bull calves. A combination of two hormones (IGF-I and FSH) enhanced multiplication of Sertoli cells. We concluded that increased IGF-I concentrations in bull calves fed high-levels of nutrition during early life increases Sertoli cell number, supports testicular development and results in earlier puberty.
RD16190Effects of freezing and activation on membrane quality and DNA damage in Xenopus tropicalis and Xenopus laevis spermatozoa
Xenopus frogs are key organisms for studying basic biology and for modelling human disease. Genetically altered lines of frogs are therefore made, and it is important for animal welfare and cost-effectiveness to store these as frozen spermatozoa. In this study we show that Xenopus sperm cryopreservation damages the DNA and plasma membrane of spermatozoa and start to identify factors affecting damage levels.
RD15553In vivo and in vitro maturation of rabbit oocytes differently affects the gene expression profile, mitochondrial distribution, apoptosis and early embryo development
Understanding the cellular and molecular mechanisms implicated in oocyte competence during rabbit in vivo and in vitro maturation is necessary to improve IVM systems in these species. This study showed for the first time that gene expression profile of in vivo-matured rabbit COCs is different from that of IVM COCs and these changes are clearly accompanied by stage-specific cytoplasmic remodelling of mitochondria, a lower rate of apoptosis of in vivo-matured CCs and better early embryo development, indicating that oocyte quality is compromised during IVM.
RD16269Sperm antioxidant defences decrease during epididymal transit from caput to cauda in parallel with increases in epididymal fluid in the goat (Capra hircus)
The antioxidant defences of epididymal fluid and spermatozoa are modulated from the caput to cauda epididymidis in a region-specific manner in the goat (Capra hircus), and loss of some of the antioxidant enzymes from spermatozoa during their epididymal transit is compensated for through a simultaneous increase in the expression and/or activity of these enzymes in the epididymal fluid of the respective segments.
Reports of the effects of birthweight on testicular development and implication for sperm production in boars are scarce. These effects were investigated in this study. It seems that high birthweight boars have the potential to produce more spermatozoa and semen doses per ejaculate. Hence, the selection of those boars would be predictive of better lifetime productivity in the boar stud.
RD16135Fish oil and wheat-germ oil supplementation restores ovarian function in streptozotocin-diabetic rats
Diabetes has a profound impact on women’s reproductive health. We aimed to investigate the protective effect of fish oil and wheat-germ oil supplementation on ovarian function in streptozotocin-diabetic rats. These natural supplements counteracted oxidative stress and inflammation induced by diabetes and regained ovarian function by increasing FSH, E2, LH and AMH levels and restoring follicle counts.
Several voltage-gated K+ (Kv) channel subunits are expressed in testis, but their contribution to testicular function remains largely unknown. We provide direct evidence of the relevance of Kv6.4 for male fertility since the targeted deletion of this gene in mice disrupted spermiogenesis, resulting in male sterility. This makes Kv6.4 a potentially interesting pharmacological target for the development of non-hormonal male contraceptives.
RD16063Proliferation and apoptosis processes in the seasonal testicular development of the wild Daurian ground squirrel (Citellus dauricus Brandt, 1844)
Male seasonal breeders show synchronised cycles of testicular growth and involution throughout the breeding cycle. The aim of the present study was to elucidate the regulatory role of cell proliferation and apoptosis in this dynamic testicular development. The study demonstrates, for the first time, that both cell proliferation and apoptosis are stimulated during the prehibernation period in wild Daurian ground squirrels. These findings advance our current understanding of the regulatory mechanisms of seasonal reproduction.
RD15524Analysis of in vitro follicle development during the onset of premature ovarian insufficiency in a mouse model
The developmental potential of follicles from a mouse model of premature ovarian insufficiency (POI) was assessed using follicle culture. Although, follicles cultured from 3- and 6-week-old mice had decreased survival compared to Controls, a population of follicles retain the potential of full development in vitro. These results indicate that in vitro follicle culture is a promising method to rescue follicle development after the onset of POI in this model.
Mouse oocyte maturation is a unique asymmetric division and a lot of polarity proteins may play vital roles during this progress. Our study described an evolutionarily conserved polarity protein Discs-large homologue 1 becomes highly polarized by associating with cytoskeleton and lose-of-function of Dlg1 compromises oocyte maturation. It may provide a new theoretic framework for mammalian meiosis research.
RD16238Imprinted and DNA methyltransferase gene expression in the endometrium during the pre- and peri-implantation period in cattle
A comprehensive understanding of biological processes in the endometrium is vital for identifying the key factors required for successful embryo recognition and implantation. The aim of the present study was to compare the expression of imprinted genes and DNA methyltransferases in the endometrium of cyclic and pregnant beef heifers. Results from this investigation provide new evidence that, in addition to their role in fetal and placental development, imprinted genes and DNA methyltransferases may have a role in establishing uterine receptivity.
RD15489Expression of C-type lectin receptors and Toll-like receptors in decidua of patients with unexplained recurrent spontaneous abortion
This study aimed to explore the mechanisms underlying the pathogenesis of unexplained recurrent spontaneous abortion (URSA). The results suggest that the balance between C-type lectin-like receptors and Toll-like receptors (TLRs) was tilted towards a TLR-dominant response in URSA patients, which may disrupt maternal–fetal immune tolerance, resulting in spontaneous abortion.
MicroRNAs (miRNAs) control several reproductive functions, including oocyte maturation, implantation and early embryonic development. We developed a method for accurate miRNA identification in a small sample of bovine blastocysts. Using pools of only 30 embryos, we identified miRNAs that regulate target genes and metabolic pathways that had not been associated with early embryogenesis before, and are potential biomarkers of developmental competence.
RD15514Role of insulin-like growth factor 1 on cross-bred Bos indicus cattle germinal vesicle oocytes exposed to heat shock
Germinal vesicle oocytes are susceptible to heat stress. The present study demonstrated that the physiological IGF-I concentration of 12.5 ng/ml was able to revert the deleterious effects of heat shock on bovine GV oocyte cellular function and ability to became a blastocyst. This thermoprotective role of IGF1 warrants further in vivo IGF1 manipulation to improve dairy cow fertility during summer.
RD15488In vitro oocyte maturation in a medium containing reduced sodium chloride improves the developmental competence of pig oocytes after parthenogenesis and somatic cell nuclear transfer
The quality of oocytes is a crucial factor in the field of reproductive biotechnology. We observed that maturation of pig oocytes in hypotonic medium with reduced NaCl improves the developmental competence of oocytes by affecting the cytoplasmic microenvironment. An oocyte maturation system using a medium with reduced NaCl could potentially improve the efficiency of reproduction in livestock species after further optimisation.
We recently discovered that the male seminal vesicles harbour a novel seminal fluid microbiome (SFM). The present study shows that short-term feeding of a high-fat diet (HFD) changes the bacterial composition of the seminal fluid and fecal microbiomes. HFD-induced SFM alterations might lead to detrimental health consequences in males, their reproductive partners and resulting offspring.
Serum affects lipid metabolism in oocytes and embryos, and cGMP has been shown to affect lipolysis in adipocytes. The present study shows that serum in the maturation medium interferes with lipid metabolism in oocytes and cumulus cells and these effects depend, at least in part, on the cGMP pathway. Evidence of involvement of this pathway in lipolysis in cumulus–oocyte complexes could contribute to the development of better culture systems for the in vitro production of oocytes and embryos with lower lipid accumulation and better cryotolerance.
RD15530Thawing boar semen in the presence of seminal plasma improves motility, modifies subpopulation patterns and reduces chromatin alterations
Seminal plasma could benefit boar semen when added during thawing. This study shows that that the addition of 10% or 50% seminal plasma during post-thawing incubation improved total motility and the motility pattern (motile subpopulations) of spermatozoa. Chromatin changes were almost non-existent for samples from some male boars, both after thawing and during the incubation, but in males susceptible to these sorts of chromatin changes, seminal plasma reduced or abolished them.
RD16124Periconceptional undernutrition affects in utero methyltransferase expression and steroid hormone concentrations in uterine flushings and blood plasma during the peri-implantation period in domestic pigs
A restricted diet applied to females during the peri-conceptional period may have consequences during the peri-implantation period. In this study we examined DNA methyltransferase 1 (DNMT1) and DNMT3a expression in endometrium and myometrium and oestradiol 17β (E2) and progesterone concentrations in the uterus and blood plasma in pigs during the peri-implantation period. Undernutrition during the peri-conceptional period affects maintenance and de novo DNA methylation in the endometrium, de novo DNA methylation in the myometrium and intrauterine E2 concentrations during the peri-implantation period.
RD16205Changes in testicular function proteins and sperm acrosome status in rats treated with valproic acid
The mechanism by which valproic acid (VPA) causes male infertility is not known. In the present study, testicular proteins and acrosome status were investigated in VPA-treated rats. Acrosome reacted spermatozoa and the expression of Ki-67, cholesterol side-chain cleavage enzyme (P450scc) and tyrosine phosphorylated proteins, except for steroidogenic acute regulatory protein, were decreased in VPA-treated rats. These changes to testicular proteins responsible for spermatogenesis result in male infertility after VPA treatment.
RD16163Potential risks to offspring of intrauterine exposure to maternal age-related obstetric complications
Several hypotheses have been proposed to explain the negative effects of delayed motherhood on an offspring’s morbidity later in life, but they are not supported by clinical and epidemiological evidence. In the present study, we show that the negative effects on offspring of delayed motherhood may be induced by embryo or fetal epigenetic DNA reprogramming associated with maternal age-related obstetric complications. This outcome has direct clinical implications for those women wanting to enrol in a fertility preservation program to offset age-related declines in fertility.
RD16056Evidence of proteinuria, but no other characteristics of pre-eclampsia, in relaxin-deficient mice
This paper reports that pregnant mice deficient in relaxin (Rln–/–) did not exhibit characteristics associated with preeclampsia; plasma sFlt-1 concentrations and blood pressure in pregnant Rln–/– mice were no different compared with age-matched wildtype mice. Although there was evidence of proteinuria in older pregnant Rln–/– mice, this was also observed in non-pregnant mice and was not a consequence of pregnancy. Therefore, pregnant Rln–/– mice are not a suitable animal model to study preeclampsia.
RD16007Circumventing the natural, frequent oestrogen waves of the female cheetah (Acinonyx jubatus) using oral progestin (Altrenogest)
We found that cheetahs do not display typical estrous cycles, but rather frequent waves of estrogenic activity every 7 to 10 days. For effective ovulation induction and the most mature oocytes, gonadotropin treatments had to be given at least 3 days after an estrogen peak. Follicular activity can be suppressed by progestin supplementation, thereby providing a way to quiesce the ovary beforehand.
RD15463Beneficial effects of diazepin-quinazolin-amine derivative (BIX-01294) on preimplantation development and molecular characteristics of cloned mouse embryos
Somatic cell nuclear transfer is frequently associated with abnormal epigenetic modifications. The present study investigated the effects of BIX-01294 on cloned mouse preimplantation embryos and the results indicated that BIX-01294 treatment of cloned embryos has beneficial effects in terms of correcting abnormal epigenetic modifications. It will be investigated further whether BIX may affect postimplantation development by correcting epigenetic modifications.
RD15546Expression of macrophage migration inhibitory factor (MIF) in bovine oviducts is higher in the postovulatory phase than during the oestrus and luteal phase
Macrophage migration inhibitory factor (MIF) may be the molecular link between maternal obesity and the quality of early embryos. This study revealed that MIF expression in the oviduct, where mammalian embryos spend their first 3 days, is higher during the postovulatory phase than during oestrus and the luteal phase in heifers. Therefore, oviduct MIF may be essential for producing good-quality embryos.
RD15539Stathmin 1 plays a role in endometrial decidualisation by regulating hypoxia inducible factor-1α and vascular endothelial growth factor during embryo implantation
The aim of this study was to explore the mechanism of action of stathmin 1 (Stmn1), a protein identified by two-dimensional electrophoresis (2-DE) and mass spectrometry (MS) in a previous proteomic analysis, in regulating embryo implantation, a critical process in pregnancy. Stmn1 was found to play a role in decidualisation and embryo implantation by inhibiting the expression of phosphorylated Akt, hypoxia-inducible factor-1α and vascular endothelial growth factor. These findings contribute to a better understanding of the potential role of Stmn1 during embryo implantation.
This review describes the involvement of the sodium pump Na+/K+-ATPase in various aspects of animal reproduction, including critical roles in embryonic development and specific sperm functions. In addition to regulating ion concentrations, Na+/K+-ATPase participates in several cell signalling pathways. Understanding the role of this molecule creates new opportunities to enhance or suppress various aspects of reproductive biology.
RD16031Methylated oligonucleotide (MON)-induced promoter hypermethylation is associated with repression of CDH1 expression and contributes to the migration and invasion of human trophoblast cell lines
Aberrant DNA methylation has been linked to deficiencies in trophoblast motility and invasion in pregnancy complications. We increased trophoblast cell motility and invasiveness through inducing site-specific hypermethylation and repressing gene expression of CDH1 by sense-methylated oligonucleotides (MONs). The results indicate that MON-mediated DNA methylation on the CDH1 promoter may alter gene expression and contribute to trophoblast motility and invasion.
The use of sex-sorted fresh spermatozoa has the potential to yield higher pregnancy rates than sex-sorted frozen spermatozoa. This study sought to characterise the in vitro quality of X and Y sex-sorted bull spermatozoa and identified the need for tailored diluents for fresh sex-sorted spermatozoa to avoid head-to-head agglutination. The use of fresh sex-sorted spermatozoa, when managed correctly, has enormous potential in the dairy industry.
This work demonstrates that it is necessary for progressive sperm motility that flagellar proteins, like ODF1, maintain reduced cysteines after epididymal transit. To determine how oxidative changes in flagellar proteins affect sperm motility, a sulfhydryl blocker was used; sperm progressive motility was abolished without affecting the level of ATP. Incomplete oxidation in flagellar proteins of asthenozoospermic samples was also observed. Inadequate maturation or an unbalanced thiol status leads to non-progressive motility.
RD16104GRIM-19, a gene associated with retinoid-interferon-induced mortality, affects endometrial receptivity and embryo implantation
We present a new hypothesis of the molecular mechanisms responsible for implantation, which could provide an important basis for further understanding of embryo implantation. Our study shows that spontaneous elimination of GRIM-19 is required for endometrial receptivity and the embryo implantation process. Our results demonstrate the involvement of GRIM-19 in the embryo implantation process by regulating adhesion, apoptosis and immune tolerance.
Using a hamster model, we found significant changes in mitochondrial number, structure and function as well as fecundity, ova production and follicular number in aged compared to younger animals. Old hamsters had decreased fertility rates, smaller litters, and decreased numbers of preantral follicles and MII oocytes. In addition, old ova contained large areas of collapsed, non-luminal cytoplasmic lamellae. Finally, old hamster ova contained higher levels of reactive oxygen species, lower mitochondrial membrane potential and decreased mitochondrial numbers, all of which could be contributing to the age-related decline in mammalian fertility rates.
Androgens (male steroids) interact with the key regulator of female reproduction, FSH, but the nature of their interaction is not clear. Examination of mutant androgen-insensitive mice with and without the expression of human transgenic FSH (Tg-FSH) revealed that subfertility in androgen-insensitive mice is rescued by the presence of Tg-FSH. These findings demonstrate that androgens and FSH actions interact in a synergistic manner to maintain optimal female fertility.
RD16061Putrescine supplementation during in vitro maturation of aged mouse oocytes improves the quality of blastocysts
Poor egg quality underlies reproductive aging in mice and in women. Oocyte maturation is a critical stage in determining the quality of eggs and hence embryonic potential. We demonstrated that putrescine supplementation during in vitro maturation of aged mouse oocytes resulted in healthier embryos. Peri-ovulatory putrescine supplementation may be an effective therapy in reproductive aging.
RD16071Stem cell factor promotes in vitro ovarian follicle development in the domestic cat by upregulating c-kit mRNA expression and stimulating the phosphatidylinositol 3-kinase/AKT pathway
Stem cell factor (SCF) and epidermal growth factor (EGF) play key roles in ovarian folliculogenesis. Herein we report that SCF promotes cat follicle growth by upregulating c-kit mRNA expression and AKT phosphorylation. EGF suppresses the stimulating effect of SCF, leading to a downregulation of FSH receptor mRNA expression. The ability to grow premature follicles in vitro could become a pivotal tool for preserving fertility in animal disease models and endangered felids.
RD15391Iloprost supports early development of in vitro-produced porcine embryos through activation of the phosphatidylinositol 3-kinase/AKT signalling pathway
Despite evidence of the presence of PGI2 in mammalian oviducts, its role in development of early embryos is largely unknown. We revealed that supplementation of PGI2 analog enhanced early development of IVP porcine embryos through activation of AKT cascade. These results indicate that iloprost can be a useful IVC supplement for production of IVP early porcine embryos with high quality.
Diabetes mellitus (DM) has been associated with male reproductive impairment but the underlying mechanisms are poorly understood owing to the diversity of factors involved in the disease. In this study we focused on the in vitro effects of high glucose levels, the primary recognised signature of DM, on spermatogenesis. We found that while mouse testicular luminal area was decreased, Sertoli cell number was increased. These findings suggest an impairment of Sertoli cell function in high-glucose conditions that may impact spermatogenesis.
RD16098Sex of co-twin affects the in vitro developmental competence of oocytes derived from 6- to 8-week-old lambs
The effect of birth type (single, twin, triplet) and co-twin sex on in vitro developmental competence of oocytes from 6 to 8-week old lambs significantly increased blastocyst development rates for lambs born with a female co-twin compared with a male co-twin. This indicates a female co-twin enhancement effect on oocyte development. Selection of lambs for embryo production programs based on co-twin sex is warranted.
Spermatozoa maturation is a necessary step for the best success in artificial reproduction. This study focused on the role of calcium ions in the acquisition of testicular sturgeon sperm to be able to swim: our results show that an influx of Ca2+ ions is required for finalisation of this maturation process.
RD15309Maternal food restriction in rats of the F0 generation increases retroperitoneal fat, the number and size of adipocytes and induces periventricular astrogliosis in female F1 and male F2 generations
Transgenerational food restriction and overweightAdverse environmental factors during gestation, such as under- or overnutrition, may predispose individuals to certain diseases later in life by altering their fetal programming. The present study investigated whether prenatal 40% food restriction was able to induce weight, metabolic and neuroimmune disturbances in the next two generations, even if they were normally fed. We showed a transgenerational transmission effect for overweight over two generations, associated with changes in immune and inflammatory patterns, revealing the severe impact of undernutrition over generations.
RD16123Cryptic male choice: experimental evidence of sperm sex ratio and seminal fluid adjustment in relation to coital rate
Mammalian males do not produce equal numbers of X- and Y-chromosome-bearing sperm, but factors influencing the variation are unknown. We show that sperm sex ratios and ejaculate traits vary with mating rate in laboratory mice, and hypothesise that males that mate frequently are usually more attractive, and would be advantaged by producing sons that inherit their attractiveness.
RD15520Temporary undernutrition during early gestation, corpora lutea morphometrics, ovarian progesterone secretion and embryo survival in gilts
This study looked at how off-feed incidents or competition for feed can affect hormonal regulation of luteal function and early pregnancy in pigs. We found no effect of a day’s fasting on LH secretion, but progesterone secretion was reduced and fewer piglets were born at subsequent farrowing. Malnutrition in group-housed sows or in periods of hot weather can potentially reduce ovarian progesterone output and fertility.
RD15357Non-genomic action of vitamin D3 on N-methyl-D-aspartate and kainate receptor-mediated actions in juvenile gonadotrophin-releasing hormone neurons
Vitamin D deficiency has been linked to impaired pubertal development affecting reproductive physiology. The effects of vitamin D3 were investigated on the juvenile gonadotrophin-releasing hormone (GnRH) neuron, a central regulator of the hypothalamic–pituitary–gonadal axis. Exposure to vitamin D3 suppressed the N-methyl-D-aspartate and kainate-mediated responses of juvenile GnRH neurons. We suggest that vitamin D is essential for normal and timely functioning of pubertal development.
RD16077Aquaglyceroporins 3 and 7 in bull spermatozoa: identification, localisation and their relationship with sperm cryotolerance
For the first time the presence and localisation of AQP3 and AQP7 in bull spermatozoa was determined, together with their relationship to sperm cryotolerance. Our findings show that neither the relative amounts nor the localisation of these proteins was altered by cryopreservation procedures. Furthermore, the abundance of AQP7 in fresh semen was found to predict bull sperm cryotolerance.
RD15414Membrane lipid profile of in vitro-produced embryos is affected by vitrification but not by long-term dietary supplementation of polyunsaturated fatty acids for oocyte donor beef heifers
In this study we evaluated the effect of feeding oocyte donor beef heifers with a protected fat enriched in linoleic acid on embryonic developmental rates after IVF, embryo cryotolerance and the membrane lipid composition of fresh and vitrified–warmed embryos. Although embryo yield, cryotolerance and membrane lipid profile were unaffected by dietary supplementation, the membrane lipid profile was affected by the cryopreservation process. Because the lipid ion abundances of fresh and vitrified–warmed embryos were distinct, they can be used as potential markers of post-cryopreservation embryonic survival.
RD15330The effects of sildenafil citrate on feto–placental development and haemodynamics in a rabbit model of intrauterine growth restriction
Current treatments for intrauterine growth restriction are not particularly effective. Sildenafil citrate (SC), a vasodilator and angiogenic molecule, could be an option to enhance conceptus growth. Here we studied, in a rabbit model, whether the administration of SC could improve or ameliorate diet-induced defects in conceptus development and offspring outcome. SC seems to benefit pregnancies complicated by placental insufficiency; however, further research is needed.
RD15483Variation among individual bulls in the distribution of acrosomal tyrosine-phosphorylated proteins in epididymal and ejaculated spermatozoa
A valid biomarker for sperm molecular characterisation was found in Japanese black cattle. The distribution of acrosomal tyrosine-phosphorylated proteins largely differs among individual bulls. The grades of acrosomal freezability and results of AI with cryopreserved spermatozoa can be predicted by assessing the distribution of acrosomal tyrosine-phosphorylated proteins in spermatozoa before cryopreservation.
RD15538Expressions of lipoprotein receptors and cholesterol efflux regulatory proteins during luteolysis in bovine corpus luteum
Progesterone, which is essential for pregnancy in mammals, is synthesised from cholesterol in the corpus luteum (CL). We investigated the involvement of cholesterol uptake receptors and efflux transporters in bovine CL regression (luteolysis), and found that the expression of scavenger receptor B1, a cholesterol uptake receptor, decreased during luteolysis. The decline in cholesterol uptake may reduce progesterone production in the CL during luteolysis.
The effect of cytokines on human male fertility is attracting more attention. In this in vitro study, the effects of TNF-α and its antagonist, etanercept, in ejaculated human spermatozoa were analysed. The negative influence of TNF-α on sperm function and the positive action of etanercept were demonstrated. This data may have clinical relevance for the management of chronic inflammatory diseases related to male reproduction.
The occurrence of intrauterine growth restriction is linked to a plethora of non-communicable disorders during postnatal life. Our study, using a translational swine model, provides evidence that prenatal developmental is strongly modulated by fetal genotype and sex. Such findings stress the necessity for considering ethnicity and sex effects in the study of the developmental origins of adult health and disease.
RD16033Actions of activin A, connective tissue growth factor, hepatocyte growth factor and teratocarcinoma-derived growth factor 1 on the development of the bovine preimplantation embryo
The reproductive tract secretes molecules collectively known as embryokines that regulate embryonic growth and development. In this study we tested four growth factors expressed in the endometrium for actions on the bovine preimplantation embryo. All four molecules affected embryonic development or gene expression and may therefore function in the endometrium as embryokines.
RD15415Temporal regulation of extracellular signal-regulated kinase 1/2 phosphorylation, heat shock protein 70 and activating transcription factor 3 during prostaglandin F-induced luteal regression in pseudopregnant rats following heat stress
High temperatures are correlated with reduced fertility. Ovarian function in pseudopregnant rats was determined in a laboratory setting of repeated heat stress. Heat stress altered the activity of mitogen-activated protein kinases and the expression of heat shock protein 70 and activating transcription factor 3 in the corpus luteum. A deeper understanding of the effects of heat stress on ovarian function will enable better approaches to promote female fecundity and sustainable animal production.
ISG15 mRNA exists in bovine embryo from one cell stage, which is earlier than IFNT mRNA. When the expression of ISG15 was knockdown, the development of blastocyst was negatively affected, , and the expressions of Ets2, IFNT and connexion 43 were inhibited in bovine blastocysts.
RD16026p21-activated kinase 1 activity is required for histone H3 Ser10 phosphorylation and chromatin condensation in mouse oocyte meiosis
Histone H3 phosphorylation is essential for chromatin condensation, which is a prerequisite for accurate chromosome separation in mitosis or meiosis. The present study demonstrates that p21-activated kinase 1 (Pak1) activity is required for histone H3 phosphorylation and consequent chromatin condensation in mouse oocytes upon the resumption of meiotic progression.
RD16020Implantation rates subsequent to the transfer of embryos produced at different phases during double stimulation of poor ovarian responders
Feasibility of luteal phase ovarian stimulation has already be certified in poor ovarian responders. Embryos produced during the luteal phase resulted in higher implantation rates compared with those produced during the follicular phase. Luteal phase ovarian stimulation should be recommended for poor ovarian responders.
Fipronil, an insecticide widely used in agriculture, veterinary medicine and public health, has been listed as a potential endocrine disrupter. The present study showed that fipronil can affect the sperm motility and compromise sperm quality of the animals. The perinatal exposure to fipronil has long-term effects on sperm parameters, and the epididymis can be a target organ.
RD15492Identification of motility-associated progesterone-responsive differentially phosphorylated proteins
Progesterone is a steroid present in the female reproductive tract that aids sperm motility. In the present study we identified human sperm proteins that are modified in response to progesterone and propose a mechanism by which progesterone would aid motility. This information may be useful for the development of drugs to improve sperm motility and contraceptives.
RD15432Prominin-1 glycosylation changes throughout early pregnancy in uterine epithelial cells under the influence of maternal ovarian hormones
The prominin-1 protein has previously been broadly investigated for its potential role in cancer and stem cell biology. This is the first investigation into the role of prominin-1 in early pregnancy. The study localises prominin-1 to the apical region of uterine epithelial cells and shows that maternal ovarian hormones significantly affect the glycosylation and abundance of this protein.
RD15290Anti-Müllerian hormone reduces growth rate without altering follicular survival in isolated caprine preantral follicles cultured in vitro
Investigations into the relationship between FSH and anti-Müllerian hormone (AMH) during in vitro culture of isolated preantral follicles are lacking. The aim of the present study was to evaluate the effect of AMH, with and without FSH, on the in vitro development of isolated caprine follicles. We demonstrated that AMH reduces follicular growth without altering follicular survival. Thus, AMH may play an important role in regulating the growth of caprine preantral follicles.
RD15253Variable duration of reproductive suppression in male coyotes (Canis latrans) treated with a high dose of the gonadotrophin-releasing hormone agonist deslorelin
A unique approach to managing coyote (Canis latrans) depredation is the development of a one-time deliverable chemical sterilant. We hypothesised that exposure to a high dose of a gonadotrophin-releasing hormone (GnRH) agonist would cause prolonged suppression of the reproductive axis. The results of the present study suggest that a plasma concentration of 100 pg mL–1 deslorelin must be maintained and that elevations beyond this threshold are likely to be unimportant.
RD15261Papain and its inhibitor E-64 reduce camelid semen viscosity without impairing sperm function and improve post-thaw motility rates
Sperm freezing has advanced animal breeding and genetics in numerous species but is not successful in alpacas due to the gel-like nature of the semen. This research aimed to reduce the viscosity of alpaca semen without affecting sperm function. The enzyme papain (0.1 mg mL–1 for 20 min at 37°C) followed by the papain inhibitor E-64 (10 µm for 5min at 37°C) eliminated viscosity and aided cryopreservation of alpaca spermatozoa. These findings will help to advance the use of reproductive technologies in camelids.
RD15494Optineurin suppression activates the mediators involved in the terminal effector pathways of human labour and delivery
Preventing spontaneous preterm birth, which is the leading cause of perinatal mortality and morbidity, is one of the most important issues facing perinatal medicine today. In this study, we found that spontaneous labour is associated with an increase in optineurin (OPTN) expression in fetal membranes and the myometrium, where it functions as a negative regulator of prolabour mediators. Elucidating the role of OPTN in labour processes enhances our understanding of the mechanisms that drive preterm birth and could perhaps lead to the development of a clinical therapeutic strategy.
Early development of the equine embryo occurs in the oviduct, and information on the composition of this oviducal environment is very valuable. We identified the proteins in the oviducal fluid of the horse, and proteins that are important for pregnancy were found in higher amounts when an equine embryo was present. These findings provide a solid basis for further studies on embryo–maternal interactions in the horse.
RD15474Sperm function during incubation with oestrus oviductal fluid differs in bulls with different fertility
Prediction of breeding bull fertility is of major economic value to the cattle-breeding industry. We proved that spermatozoa from high-fertile bulls showed a low degree of tyrosine phosphorylation and high degree of capacitation and acrosome reaction upon exposure to oviductal fluid. These findings may help in developing tools for assessing fertility of bulls, once validated in more animals.
RD15336Carnitine-mediated antioxidant enzyme activity and Bcl2 expression involves peroxisome proliferator-activated receptor-γ coactivator-1α in mouse testis
Because carnitine treatment is used in many cases of male infertility, the aim of the present study was to evaluate the mechanism by which carnitine improves spermatogenesis in mice. In the study, carnitine treatment of mice stimulated peroxisome proliferator-activated receptor-γ coactivator-1α expression, which, in turn, regulated steroidogenesis by enhancing expression of Bcl2 and antioxidant enzymes in the testis.
RD15361Oestrogenic action of neonatal tamoxifen on the hypothalamus and reproductive system in female mice
Tamoxifen is widely used in the treatment of breast cancer and can interfere with the reproductive system. The present study showed that tamoxifen administration during the neonatal period has oestrogenic actions on oestrogen-sensitive nuclei in the brain controlling the reproductive axis in female mice. Therefore, tamoxifen may have adverse effects on brain differentiation and reproductive function.
RD15214Prion protein 2 (dublet) gene (PRND): role in ovine semen capacitation, cryopreservation and fertility
The physiological functions of prion and prion-like proteins are overlapped by their pathological features, namely in the invariably fatal neurodegenerative diseases of both humans and animals, such as Kuru, Creutzfeldt-Jakob disease and new variant, bovine spongiform encephalopathy and sheep scrapie. The aim of the present study was to investigate the role of the prion-like Doppel (PRND) protein and different PRND genotypes in the sperm capacitation process, cryoresistance, and fertility potential. Both factors were crucial for sperm capacitation potential, cryoresistance, and embryo production. Thus, an important physiological function of Doppel protein and genotype in male fertility has been demonstrated.
RD15485Short- and long-term outcomes of the absence of protein during bovine blastocyst formation in vitro
Removal of protein from culture medium improves analytical reliability, although bovine embryo development in vitro could be affected. However, protein removal over a 24-h period in culture led to embryos with reduced miscarriage rates without affecting birthweight, gestation length or calf morphology. This finding represents direct access to culture medium as a substrate for non-invasive analysis, leading to potential biomarker discovery without compromising embryonic viability.
RD15440Effect of Single Layer Centrifugation on reactive oxygen species and sperm mitochondrial membrane potential in cooled stallion semen
Stallion spermatozoa lose viability and fertilising capability relatively quickly when cooled; robust spermatozoa can be selected by colloid centrifugation. In the present study, selected spermatozoa were shown to metabolise differently to controls, producing less hydrogen peroxide, which is known to damage spermatozoa. These results indicate that colloid centrifugation could be used to improve the quality of stallion semen for AI.
Cloned concepti on Day 225 of gestation were characterised by lower pregnancy rates, high gestational losses, a distinctive biphasic growth pattern, with early growth restriction followed by accelerated growth and foetal weight on Day 225, and significant morphological changes in placental mass and shape when compared with in vivo-derived pregnancies. Conversely, Day-225 IVF-derived concepti were widely similar to controls.
Prostaglandins induce contraction and relaxation of oviductal smooth muscle, which are essential for transport of gametes and embryo. The present results demonstrate that lysophosphatidic acid significantly stimulated prostaglandin production by cultured isthmic stromal cells. It is suggested that locally produced lysophosphatidic acid contributes to oviductal motility in the isthmus by regulating prostaglandin production in cows.
RD15306Increased frequency of chromosome congression defects and aneuploidy in mouse oocytes cultured at lower temperature
In vitro manipulation of oocytes is a standard procedure used in assisted reproduction. The aim of the present study was to analyse the effect of suboptimal culture temperatures on various aspects of meiotic division, including chromosomal congression and segregation defects. The results revealed that even a small deviation from optimal culture temperature has a marked effect on chromosome congression and aneuploidy.
RD15424Lower dietary n-6:n-3 ratio and high-dose vitamin E supplementation improve sperm morphology and oxidative stress in boars
Dietary PUFA and vitamin E are known to influence sperm morphology and oxidative stress. The effects of dietary n-6 : n-3 and vitamin E on the sperm morphology and oxidative stress in boars were investigated. An n-6 : n-3 ratio of 6 :1 and 400 mg kg–1 vitamin E have beneficial effects on sperm morphology by improving oxidative stress.
RD15430Expression of 17β-hydroxysteroid dehydrogenase and the effects of LH, FSH and prolactin on oestrone and 17β-oestradiol secretion in the endometrium of pigs during early pregnancy and the oestrous cycle
The uterus produces steroids and primary steroidogenic enzymes are expressed in the uterus of pregnant and cyclic pigs. In this study we examined 17β-hydroxysteroid dehydrogenase expression and the effects of LH, FSH and prolactin on oestrone and 17β-oestradiol release in vitro in endometrial explants. We show that LH, FSH and prolactin may regulate oestrogen release by the endometrium.
RD15390Immunolocalisation of aromatase regulators liver kinase B1, phosphorylated AMP-activated protein kinase and cAMP response element-binding protein-regulated transcription co-activators in the human testis
The hormone oestrogen is essential for testis development and its synthesis is dependent on the enzyme aromatase. However, the molecular mechanism of aromatase regulation is still poorly understood. This study describes for the first time the cellular localisation of proteins important for aromatase regulation, which are well known in breast tissue, in the normal testis. This new finding may help develop new therapies for oestrogen-mediated infertility.
RD15378Lonidamine-ethyl ester-mediated remodelling of the Sertoli cell cytoskeleton induces phosphorylation of plakoglobin and promotes its interaction with ?-catenin at the blood–testis barrier
We are in need of safe, effective and affordable male contraceptives. Indazole carboxylic acids such as lonidamine and adjudin disrupt adhesion between Sertoli and germ cells in the rat testis, resulting in infertility. One of the key findings of this study was that lonidamine-ethyl ester also displayed contraceptive effects. Additional studies are required to determine whether indazole carboxylic acids can enter the clinical pipeline
For the first time, a minimal invasive method that allows ovarian tissue collection from sows is described in detail. This method, a transvaginal ultrasound-guided biopsy, is quick, can be performed by one person, and has no major effects on the sow’s reproductive performance. It can be applied on the farm to study effects of housing or nutrition on luteal function.
RD15315Insulin exposure during in vitro bovine oocyte maturation changes blastocyst gene expression and developmental potential
Metabolic imbalance impairs female fertility. We wanted to find out how metabolism affects bovine oocyte quality, and demonstrated that insulin exposure during in vitro oocyte maturation affects embryo development, morphology and gene expression. Even short-term exposure to a metabolic imbalance has lasting effects on embryo development, and this metabolic programming may have consequences later in life.
RD15425Greater sperm complexity in the Australasian old endemic rodents (Tribe: Hydromyini) is associated with increased levels of inter-male sperm competition
Most species of Australasian old endemic rodents have a highly complex sperm head morphology that contains an apical hook together with two ventral processes of variable length and orientation. This study tested whether this variation is a sexually selected trait using relative testes mass as a proxy for the intensity of inter-male sperm competition and controlling for phylogeny. The angle of these processes, but not length, was positively correlated with relative testes mass; its possible functional significance is briefly discussed.
RD15319Fateful triad of reactive oxygen species, mitochondrial dysfunction and lipid accumulation is associated with expression outline of the AMP-activated protein kinase pathway in bovine blastocysts
In vitro-derived bovine embryos frequently exhibit lipid accumulation, mitochondrial dysfunction and enhanced levels of reactive oxygen species. With the aim of unravelling the role of the AMP-activated protein kinase cytalytic subunit 1alpha (AMPKα), the present study identified AMPKα activity and distinct expression patterns of AMPKα-related genes to be correlated with that phenotype. These results prove the important role of AMPKα in embryonic energy metabolism.
RD15321Detection of cross-sex chimerism in the common marmoset monkey (Callithrix jacchus) in interphase cells using fluorescence in situ hybridisation probes specific for the marmoset X and Y chromosomes
Bone marrow chimerism is a unique and normal feature of Callithrichidae development that originates through shared placental circulation. A new X and Y chromosome-specific fluorescent in situ hybridisation protocol for Callithrix jacchus has been developed and validated to detect and investigate chimerism between male and female littermates. One of the strengths of this method is that it is useful in interphase cells, and allows cell morphology to remain intact to facilitate identification of cell type.
RD15358Dietary-induced gestational iron deficiency inhibits postnatal tissue iron delivery and postpones the cessation of active nephrogenesis in rats
Gestational iron deficiency can cause developmental programming of adult hypertension, but the critical developmental window during nephrogenesis has not been studied. The present study examined renal development during perinatal iron deficiency in rats and found altered timing of kidney maturation and histological renal adaptations that may contribute to dysfunctional glomeruli and renal hyperfiltration, which themselves contribute to hypertension.
Finding novel biomarkers of ovarian follicle function would be of benefit. This study investigated the follicle biomarker potential of microRNAs (miRNAs) and found that levels of miR-873 and miR-202 can be used as indicators of steroidogenic capacity in cattle. These miRNAs, in addition or alternatively to commonly used steroid level measurements, could be used to accurately classify follicles based on steroidogenic capacity.
Information about follicle density is critical to determine the quantity of fragments during biopsy procedures in women and domestic animals. This study evaluated the effects of age and determined the minimum number of ovarian fragments and histological sections required to estimate equine follicle density in ovarian biopsy fragments. The results show that follicle density is negatively influenced by increasing age and, independent of follicle heterogeneity, three to four ovarian fragments combined with 65 histological sections are required to accurately determine follicle density in equine ovarian tissue.
RD15356Glucocorticoid-induced changes in glucocorticoid receptor mRNA and protein expression in the human placenta as a potential factor for altering fetal growth and development
Glucocorticoids (GCs) sustain life and the development of human embryo; however, fetoplacental exposure to high maternal GC levels may lead to placental and fetal abnormalities, with the adverse consequences of GC excess manifested later in life. The mechanisms through which GC excess alters placental and fetal development are yet to be identified; however, changes in placental GR levels and subcellular localization and ratios may modify placental function, in utero growth of the fetus and lifelong health.
These articles have been peer reviewed and accepted for publication. They are still in production and have not been edited, so may differ from the final published form.
Supplementation of culture medium with L-carnitine improves the development and cryotolerance of in vitro produced porcine embryos
Development of Pre-implantation Porcine Blastocysts cultured within Alginate Hydrogel Systems either supplemented with Secreted Phosphoprotein 1 or conjugated with Arg-Gly-Asp Peptide
Comparative analysis of granulosa cellsâ gene expression in superstimulated Holstein cows with sub-optimal embryo yield
Andrographolide disrupts meiotic maturation by blocking cytoskeletal reorganization and decreases the fertilization potential of mouse oocytes
Melatonin potentially acts directly on swine ovary by modulating granulosa cell function and angiogenesis
Differential effects of high and low glucose concentrations during lipolysis-like conditions on bovine in vitro oocyte quality, metabolism and subsequent embryo development
Sperm motility activation in the critically endangered booroolong frog: the effect of medium osmolality and phosphodiesterase-inhibitors
Low doses of Bisphenol A can impair postnatal testicular development directly, without affecting hormonal or oxidative stress levels
Steroid hormones interact with natriuretic peptide type C to delay nuclear maturation, to maintain oocyte-cumulus communication and to improve quality of in vitro produced embryos in cattle
Glutamine protects rabbit spermatozoa against oxidative stress via glutathione synthesis during cryopreservation
Neuro-modulating effect of oestradiol in the metabolism of ovarian progesterone and oestradiol on dioestrus II day. Participation of the superior mesenteric ganglion
Endogenous lysophosphatidic acid participates in vascularization and decidualization at the maternal-fetal interface in the rat
Cerium oxide nanoparticles (CeO2 NPs) improve the developmental competence of in vitro matured prepubertal ovine oocytes
Time within reproductive season, but not age or inbreeding coefficient, influences seminal and sperm quality in the whooping crane (Grus americana)
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