Sex-specific differences have been reported in most of the diseases. This review provides data on differential adaptation of male and female fetuses in utero in response to maternal nutritional, physiological and environmental insults resulting in sex-specific differences. Because sex differences are associated with fetal outcome and survival, it is of importance to study the underlying biological mechanisms to improve both pre-natal and neonatal care.
Reproduction, Fertility and Development
Volume 29 Number 11 2017
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.
RD16235Endogenous lysophosphatidic acid participates in vascularisation and decidualisation at the maternal–fetal interface in the rat
Decidualisation and vascular remodelling are fundamental for placentation and pregnancy success. Disruption of endogenous lysophosphatidic acid signalling modified the development of uterine vessels with consequences in the formation of the decidua and placenta, compromising the growth of embryos. Defects in these mechanisms could appear as obstetric complications, like implantation failure, positioning lysophosphatidic acid as a regulatory key lipid at implantation.
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.
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.
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.
Endo-siRNAs play an important role in mouse oocyte meiosis but the function of endo-siRNAs in other species is unknown. In this study, we found that an endo-siRNA deficiency resulted in oocyte spindle abnormalities and apoptosis, suggesting that endo-siRNAs are indispensible for oocyte maturation in pigs. This study unequivocally demonstrates an essential function for endo-siRNAs in mammals.
RD16378Neuromodulatory effect of oestradiol in the metabolism of ovarian progesterone and oestradiol during dioestrus II: participation of the superior mesenteric ganglion
This work demonstrates that estradiol through the peripheral nervous system modulates the ovarian physiology, favoring the physiological luteolysis. The objective of this work was to demonstrate whether estradiol in superior mesenteric ganglion modifies the release of ovarian progesterone, estradiol, nitric oxide and noradrenaline on DII day. The ganglionic stimulation produces a decrease in the ovarian release of progesterone and estradiol by an increase of nitric oxide and decrease of noradrenaline.
RD17020Glutamine protects rabbit spermatozoa against oxidative stress via glutathione synthesis during cryopreservation
The addition of 20 mM glutamine to the freezing extender significantly improved post-thaw sperm motility, acrosome integrity, membrane integrity and mitochondrial activity. The exposure of spermatozoa to glutamine enhanced their γ-GCS activity and GSH synthesis, while decreasing ROS accumulation, LPO and DNA damage. Therefore, glutamine protects spermatozoa against ROS-induced cryodamage by enhancing GSH synthesis.
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.
RD16289Successful chimera production in the Hungarian goose (Anser anser domestica) by intracardiac injection of blastodermal cells in 3-day-old embryos
The aim of this study was to develop a new genome preservation technique for goose species. According to our results, goose primordial germ cells (PGCs) migrate between 69 and 84 h of development. The goose blastoderm contains PGCs and by isolating and injecting these blastodermal cells we were able to generate chimeras. Altogether, this method could contribute to the cryopreservation of this species.
RD16320Steroid hormones interact with natriuretic peptide C to delay nuclear maturation, to maintain oocyte–cumulus communication and to improve the quality of in vitro-produced embryos in cattle
This study provides evidence that natriuretic peptide C and steroids interact to slow oocyte nuclear maturation and to enhance oocyte-cumulus communication in cattle. Moreover, it indicates that exposure of the cumulus–oocyte complex to physiological concentrations of these hormones is beneficial to oocyte developmental competence. Therefore, the dataset is of relevance for the progress of oocyte biology and for the improvement of in vitro maturation outcomes.
This paper describes findings during a unique opportunity to monitor placental structure and function throughout pregnancy in the under-researched zebra. Zebras are more donkey-like than horse-like in placental structure, rate of fetal development and gestation length. The results highlight the retention of unique reproductive mechanisms across the Genus Equus despite many other evolutionary changes within the member species.
A combination of permeating and non-permeating cryoprotectants and freezing rates was used to successfully cryopreserve saltwater crocodile spermatozoa. Although glycerol concentrations >1.35 M had an inhibitory effect on motility before freezing, 0.68 M glycerol combined with 0.2 M sucrose and post-thaw washing of spermatozoa in capacitating media resulted in motility of between 12% and 14.2%, the highest ever recorded for this species.
RD16432Low doses of bisphenol A can impair postnatal testicular development directly, without affecting hormonal or oxidative stress levels
The peripubertal period is considered a critical stage of reproductive development and more vulnerable to the action of toxic agents, such as BPA, which can lead to impaired sexual development. We investigated the exposure to BPA during the peripubertal period and eEvidencing a reduction in the number of Sertoli cells, histopathological abnormalities and changes in sperm morphology. Therefore, BPA may silently impair testicular postnatal development in rats.
High embryo loss during early pregnancy in pigs is still an unresolved issue. Comparing the proteomic profile of the endometrium harvested from crucial stages of early pregnancy (Days 12–13 and 15–16) with the proteomic profile of tissue from corresponding days of the oestrous cycle revealed the presence of proteins unique for early pregnancy that are involved in adhesion, angiogenesis and cytoskeletal organization. The results of the present study may help us understand the causes of early pregnancy loss in pigs.
Pannexins (Panx) are proteins that form functional single membrane channels that have not yet been described in dogs. The aim of the present study was to detect Panx1, Panx2 and Panx3 in frozen–thawed dog semen. All three Panx channels were present in dog spermatozoa and increased propidium iodide (PI) permeability in frozen–thawed dog spermatozoa, suggesting that the percentage of PI-positive spermatozoa used as an indicator of non-viable cells may lead to overestimation of non-viable cells.
RD17012Sperm motility activation in the critically endangered booroolong frog: the effect of medium osmolality and phosphodiesterase inhibitors
Successful artificial fertilisation primarily depends on two characteristics of male quality: 1) the maintenance of high sperm viability during storage and 2) the effective activation of sperm motility. This study identified optimal conditions for sperm motility activation in the critically endangered booroolong frog. Results reported make a significant contribution to the growing field of amphibian reproduction technologies.
RD16474Differential effects of high and low glucose concentrations during lipolysis-like conditions on bovine in vitro oocyte quality, metabolism and subsequent embryo development
Maternal metabolic disorders (e.g. obesity and diabetes in women or a negative energy balance in dairy cows) are associated with altered metabolic profiles, which can impact oocyte quality and thus fertility. We showed that altered glucose can differentially affect oocyte quality in the presence of elevated free fatty acids. These results further highlight the importance of maternal metabolic health for fertility.
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RD16309The influence of dopaminergic system inhibition on biosynthesis of gonadotrophin-releasing hormone (GnRH) and GnRH receptor in anoestrous sheep; hierarchical role of kisspeptin and RFamide-related peptide-3 (RFRP-3)
This research provides new insight into central molecular mechanisms regulating GnRH/LH secretion in anestrous sheep. To better explain this problem, herein, we demonstrated that RF-amide peptides are involved in dopamine-induced inhibition of GnRH/LH secretion. Studies expanded an important issue in the field of neuroendocrinology of reproduction, especially in cognitive appraisals of mechanisms participating in physiological regulation of GnRH/LH biosynthesis.
RD17184Expression of vascular endothelial growth factor A isoforms is dysregulated in women with endometriosis
Angiogenesis is a vital process in the development and survival of lesions during endometriosis. In the present study we identified upregulation of several isoforms of vascular endothelial growth factor (VEGF) A in the endometrium of women with endometriosis during menstruation and, for the first time, identified the natural expression of the VEGF111 isoform in humans. These findings contribute to our understanding of how the endometrium differs in women with endometriosis, and indicates a dysregulation at the critical time of menstruation.
Do extracellular vesicles have a biological role in the communication between the embryo and its microenvironment during in vitro culture? In this study we showed that embryos were able to incorporate microvesicles obtained from endometrium and amnion, but only amniotic progenitor cell-derived microvesicles resulted in better blastocyst production rate and quality compared to control. This study could be a useful starting point for further studies related to paracrine mechanisms of communication between embryos and the culture medium.
The role of melatonin as an endocrine regulator of reproductive activity is well known; however, whether it stimulates or inhibits reproductive function varies across species. We demonstrated that in vitro exposure to melatonin reduced steroid production by ovarian theca cells in cattle, implicating melatonin as a regulator of reproductive function in cattle. Because reduced reproductive capacity leads to economic losses, an understanding of the effects of melatonin on ovarian function may lead to new ways to increase reproductive efficiency in cattle.
RD17024Phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) and Janus kinase/signal transducer and activator of transcription (JAK/STAT) follicular signalling is conserved in the mare ovary
Signalling pathways responsible for follicular activation in the mare remain uncharacterised. Messenger RNA expression and protein localisation of members of the PI3K/AKT and JAK/STAT pathways were analysed using tissue from fetal and adult mare ovaries. Findings suggest that members of these pathways are indeed present and may serve as useful biomarkers for assessment of ovary development in the horse.
RD17241Effect of lipopolysaccharide on the expression of inflammatory mRNAs and microRNAs in the mouse oviduct
Uncontrolled inflammation of the oviduct can result in tubal occlusion and infertility. This inflammation is often in response to infection with sexually transmitted bacterial pathogens. This study describes how the endotoxin from Escherichia coli-derived lipopolysaccharide affects the expression of inflammatory mRNAs and post-transcriptionally modifying miRNAs within the oviduct, providing novel insight into the molecular regulation of inflammation within this key reproductive organ.
RD17076Modification of membrane cholesterol and desmosterol in chicken spermatozoa improves post-thaw survival and prevents impairment of sperm function after cryopreservation
During cryopreservation, spermatozoa undergo cryodamage because of membrane changes, including sterol loss, which results in diminished post-thaw fertilisation ability. The present study showed that loading of cholesterol and desmosterol into chicken spermatozoa prior to cryopreservation enhanced their post-thaw quality by inhibiting apoptotic changes and premature acrosome reaction, providing new mechanistic insight into cryodamage in chicken spermatozoa.
RD17049Identification and expression of forkhead box genes in the Chinese giant salamander Andrias davidianus
The forkhead box (Fox) gene family encodes multiple forkhead proteins that exhibit marked functional diversity and are involved in biological processes, especially sex differentiation. The identification, characterisation, selective pressure, protein structure and expression pattern of Fox genes in response to temperature were investigated in the present study in the Chinese giant salamander Andrias davidianus to enrich the sexual development and reproduction mechanism in amphibian.
To improve current infertility treatments, it is necessary to evaluate uterine receptivity in each menstrual cycle. If this can be evaluated beforehand, when uterine receptivity is inadequate eggs could be stored using cryopreservation. In this study in a mouse model, we identified an electrophysiological parameter that can evaluate uterine receptivity prospectively.
Kisspeptin peptide is a major regulator of initiation of pubertal actions in the central nervous system, but its development in growing animals is obscure. The presence of kisspeptin neurons in the brain from birth to puberty in relation to luteinising hormone release pattern was analysed in female lambs. Activity of kisspeptin neurons increased gradually parallel with augmentation of luteinising hormone release and reached maturation several weeks before puberty. The participation of kisspeptin in the development of the ovine reproductive processes is suggested.
RD17255Systemic adiponectin treatment reverses polycystic ovary syndrome-like features in an animal model
Adiponectin may be a potent therapeutic agent for recovery of the polycystic ovary syndrome (PCOS) condition. The aim of the present study was to unravel the role of adiponectin in regulating reproduction, metabolism and fertility status in mice in which PCOS had been induced. Reversal of PCOS-like features following adiponectin treatment in mice is a major breakthrough with regard to the effective management of PCOS.
RD16486Insulinotropic nucleobindin-2/nesfatin-1 is dynamically expressed in the haemochorial mouse and human placenta
The placenta is a multifunctional link between mother and fetus vital for nutrient transport. In this study we determined when and where nucleobindin-2/nesfatin-1 were located in the placenta and found them particularly in secretory trophoblast cells of the mouse and human placenta. The placenta should be considered a source for nucleobindin-2/nesfatin-1 during pregnancy, with possible roles in maternal glucose control and nutrient sensing.
RD17044Association of the cysteine-rich secretory protein-3 (CRISP-3) and some of its polymorphisms with the quality of cryopreserved stallion semen
Some genes are involved in stallion semen tolerance to cryopreservation. This work evaluated the relationship between CRISP-3 protein and some of its single nucleotide polymorphisms, with post-thawing semen quality in stallions. This study demonstrated that different parameters of seminal quality are influenced by the CRISP-3 genotype and the concentration of this protein in seminal plasma.
RD17094Milk-fat globule epidermal growth factor 8 (MFGE8) is expressed at the embryo– and fetal–maternal interface in equine pregnancy
Milk fat globule-EGF factor 8 protein (MFGE8) is a protein that promotes cell to cell adhesion and is thought to play an essential role in embryo implantation in mice and humans. We have described the expression of MFGE8 in the equine uterus and found particular strong expression of the protein by the fetal portion of the placenta. These results imply a role for MFGE8 in maintaining an intact placenta during mare pregnancy.
RD17095Mouse double minute homologue 2 (MDM2) downregulation by miR-661 impairs human endometrial epithelial cell adhesive capacity
A major limitation of IVF success is failure of the embryo to successfully attach to the womb. We have identified a critical new mechanism of communication between the human embryo and endometrium (inner lining of the uterus). Overall it is likely that this bi-directional communication facilitates the critical adhesion process during the initiation of successful pregnancy.
RD17146Spermatogenic phenotype of testis-specific protein, Y-encoded, 1 (TSPY1) dosage deficiency is independent of variations in TSPY-like 1 (TSPYL1) and TSPY-like 5 (TSPYL5): a case-control study in a Han Chinese population
The testis-specific protein, Y-encoded, 1 (TSPY1) gene is a newly confirmed regulator of sperm production, and dosage deficiency of the multicopy gene confers increased risk of spermatogenic failure. The aim of the present study was to evaluate whether the autosomal homologous genes originating from TSPY1 modify the spermatogenic phenotype of TSPY1. The results suggest that phenotypic expression of TSPY1 defects is independent of variations TSPY-like 1 (TSPYL1) and TSPY-like 5 (TSPYL5), highlighting the significance of TSPY1 in the modulation of spermatogenic efficiency.
The types and copy numbers of very short so-called miRNA molecules were measured in eggs and embryos of cattle. Results highlight the importance of a non-canonical miRNA during stages before embryonic transcription begins, the influence of the parental origin of miRNAs and the importance of measuring absolute as opposed to relative levels of these molecules.
RD16399Production of inbred offspring by intracytoplasmic sperm injection of oocytes from juvenile female mice
Intracytoplasmic sperm injection (ICSI) has been successfully applied to hybrid mice, but not to inbred mice due to low efficiency. Adding an activation step to ICSI of oocytes from juvenile mice results in the birth of healthy inbred pups. The results of this study show that the modified protocol can potentially improve the efficiency of ICSI in inbred mice.
A prolonged inflammatory response to breeding severely decreases fertility in horses. The aim of the present study was to evaluate differences between fertile and subfertile mares in the expression of selected genes associated with the immune response. Three of the genes analysed proved to be very sensitive and specific diagnostic markers predicting prolonged inflammation, and likely reduced fertility, prior to breeding.
Genetic background can help to explain idiopathic cases of recurrent pregnancy loss (RPL), which make up about 50% of total cases. Here, we have evaluated the genetic profiles of women experiencing RPL by analysing polymorphisms in the NOS2, PTGS2 and VEGFA genes. We found a higher incidence of a polymorphic variant in the NOS2 gene in the RPL group, suggesting a role for this gene in RPL risk, possibly due to oxidative stress mechanisms.
RD17037Oral propylene glycol modifies follicular fluid and gene expression profiles in cumulus–oocyte complexes and embryos in feed-restricted heifers
Ovum pick-up and in vitro embryo production can be used in dairy heifers. However, production results are often disappointing when animals are underfed. A short-term dietary supplement with propylene glycol (400 mL of PG or water/drench, daily drenching at 1600 hours for the first 9 days of the oestrous cycle), given to feed-restricted heifers, was able to restore the expression of selected genes in cumulus cells and blastocysts to levels seen in correctly fed heifers.
One of the predominant causes of anoestrus is a low level of ovarian oestrogens. Herein, we investigated the association of the oestrogen biosynthesis regulator CYP19A1 with the risk of anoestrus in buffaloes and found three SNPs. The risk alleles in these SNPs were significantly associated with lower levels of estradiol and antioxidants and higher incidence of oxidative ovarian damage and anoestrus.
RD16518Breeding in the fat-tailed dunnart following ovarian suppression with the gonadotrophin-releasing hormone agonist Lucrin® Depot
The application of genome storage in conservation is severely constrained by our limited access to reliable assisted breeding techniques. This is particularly so for marsupials, in which the corpus luteum, and thus cycling, is not affected by agents used to control ovarian function in eutherian mammals. Herein we report progress on the use of a single-treatment long-acting suppressor of pituitary function to reset cycling and ovulation in a marsupial.
RD17180Can resveratrol attenuate testicular damage in neonatal and adult rats exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin during gestation?
Humans may be exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), an environmental pollutant, via contaminated food, inhalation or skin contact. The effects of TCDD are exerted via binding to aryl hydrocarbon receptors (AhR). In this study we investigated the possible protective effect of resveratrol, an AhR antagonist, against testicular damage caused by TCDD exposure during pregnancy. The findings suggest that TCDD in combination with resveratrol results in more severe toxicity than administration of either drug alone.
RD17051Development of fresh and vitrified agouti ovarian tissue after xenografting to ovariectomised severe combined immunodeficiency (SCID) mice
For wild rodents such as the agoutis, the combination of gonadal tissue cryopreservation and xenotransplantation could help promote germplasm conservation and reproduction, especially in the case of genetically valuable individuals. The present study demonstrates that xenografted agouti ovarian tissue, fresh or vitrified, is able to promote the return of ovarian activity in ovariectomised SCID C57B1/6 mice.
RD16345Changes in urinary androgen concentration indicate that male giant pandas (Ailuropoda melanoleuca) respond to impending female oestrus during and outside the typical spring breeding season
Although giant pandas have been described as mono-oestrus spring breeders, males exposed to oestrous females outside this timeframe exhibit breeding behaviours and interest in mating. Urinary androgens and sperm parameters were quantified for males exposed to females expressing oestrus during spring, autumn, or winter. The data from this study showed a rapid physiological readiness of male giant pandas to mate in response to female oestrous cues within or outside of the normal breeding season and may suggest a facultative seasonal reproduction with a “female-induced rut”.
Reduced fetal growth can contribute to a greater risk of later metabolic disease, but the key mechanisms remain unknown. Herein we report findings from a large animal model (sheep) in which fetal growth was restricted by reducing maternal food intake and show that once the offspring become obese, they exhibit increased abundance of molecular markers of metabolic stress in the fat and liver. This could represent a primary mechanism by which growth-restricted offspring are at greater risk of metabolic ill health in later life.
RD16487In vitro growth and development of isolated secondary follicles from vitrified caprine ovarian cortex
In vitro growth of secondary follicles isolated from the vitrified ovarian cortex allows a better understanding of their complex development and of obtaining healthy follicles. The aim of this study was to investigate how the medium composition affects the viability and growth rate of the isolated secondary follicles. This technique could become a strategy to preserve the reproductive potential of females.
RD17217The contribution of p53 and Y chromosome long arm genes to regulation of apoptosis in mouse testis
Tumor suppressor protein p53, known widely as the guardian of genome preventing cancer formation, is also involved in physiological apoptosis of germ cells in testes. The study used a mouse model to reveal that germ cell apoptosis during the first round of spermatogenesis can be efficiently mediated by p53-independent mechanisms, but activation of these mechanisms requires engagement of multicopy Y-chromosome long arm genes. The role of Y-chromosome genes in regulation of testicular apoptosis was demonstrated for the first time.
It is not known whether female accessory reproductive organs that play a major role in reproduction are vulnerable to chronic stress effects. This study revealed irreversible oxidative stress in the Fallopian tube following short or long-term chronic stress exposure, whereas uterus changes were reversible following short-term exposure but not after long-term exposure. This study indicates that stress-induced reproductive failure may also be due to alterations in the accessory reproductive organs.
RD16536Impact of equine assisted reproductive technologies (standard embryo transfer or intracytoplasmic sperm injection (ICSI) with in vitro culture and embryo transfer) on placenta and foal morphometry and placental gene expression
Assisted reproduction techniques such as in vitro fertilisation and embryo culture have been shown to affect the size and anatomy of the newborn and the placenta in some species, but no information is available on this area in the horse. We evaluated foals and placentas from pregnancies resulting from natural conception, embryo transfer and transfer of in vitro-produced (intracytoplasmic sperm injection) embryos and found no significant differences between groups in size or weight of the foal or placenta and no differences in expression of 17 genes controlling placental function. Our findings suggest that the horse embryo differs from that of ruminants in its sensitivity to the in vitro environment, supporting the continued use of assisted reproduction techniques for commercial foal production.
The testis is the organ essential for spermatogenesis and male fertility. This paper profiles differences in DNA methylation levels during prepubertal porcine testis development, and provides new insights into mammalian testis development and represents a foundation for additional research using the pig as a model animal.
RD17041Quantitative changes of extravillous trophoblast cells in heavy smoker mothers compared with healthy controls
Extravillous trophoblast cells are one of the main elements that are affected by maternal smoking during pregnancy. Quantitative changes in extravillous trophoblast cells could be evidence for the hypothesis that changes in these cells could affect fetal health by changes in normal placentation.
RD16338Calcium-sensing receptor (CASR) is involved in porcine in vitro fertilisation and early embryo development
Fertilisation and development of the mammalian egg is still not well understood, illustrated by low efficiencies of assisted reproductive technologies. The role of a cell membrane receptor for calcium (CASR) was studied, using pig spermatozoa and eggs as models. Significant effects were demonstrated by stimulating this receptor at different steps of the process. These findings are useful for future basic and applied research.
RD17081Markers of protein synthesis are increased in fetal membranes and myometrium after human labour and delivery
Inflammation plays an important role in the initiation of preterm birth, which remains the largest cause of neonatal deaths. In the present study, components of the protein synthesis machinery were significantly increased with labouring compared to non-labouring fetal membranes and myometrium, and blockade of protein synthesis significantly reduced inflammation in the myometrium. These results may provide further insight into the pathways involved in the initiation of preterm birth.
Toll-like receptors (TLRs) contribute to an exclusive protection system in the uterus during the oestrus cycle; however, the importance of TLRs in tissue repair within the involuting uterus is not known. Herein we show that TLRs are constitutively expressed in uterine cells and that the localisation pattern of TLRs in the endometrium varies with structural changes in the involuting uterus. These results suggest that TLRs have a physiological role in the uterine remodelling events during the post partum involution period.
RD17045Unique oestrogen receptor ligand-binding domain sequence of native parrots: a possible link between phytoestrogens and breeding success
Hormones in plants may provide an important trigger for activating breeding activity in Kākāpō, a critically endangered New Zealand parrot. By comparing native and Australian parrots for their ability to respond to steroid hormones, we discovered novel sequences in the receptivity of parrots to oestrogenic compounds. Receptor modeling revealed that altered interactions with plant oestrogens in parrots may enhance the physiological pathways linked to fertility and successful breeding.
RD17068Efficient generation of goats with defined point mutation (I397V) in GDF9 through CRISPR/Cas9
Taking advantages of the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) 9 system, we achieved a precise single-nucleotide substitution in the growth differentiation factor 9 (GDF9) gene using a single-stranded oligo deoxynucleotide as a homology-dependent repair template with a relatively high efficiency in order to increase litter size in goats. The present study provides an alternative approach for introducing defined point mutations in large animal models for the improvement of significant traits, as well as for the functional validation of key variants.
RD16441Expanded equine cumulus–oocyte complexes exhibit higher meiotic competence and lower glucose consumption than compact cumulus–oocyte complexes
Equine immature oocytes are classified as expanded or compact and are matured in the laboratory to obtain offspring; higher maturation rates are reported for expanded oocytes. The aim of this study was to assess whether differences in the maturation capacity of these oocytes is related to differences in glucose uptake, metabolism or apoptosis. Consistent metabolic and genomic differences were found between expanded and compact oocytes: compact oocytes consumed more glucose but matured less than expanded oocytes. This indicates that the maturation requirements vary for compact and expanded oocytes.
RD17174Linoleic (LA) and linolenic (ALA) acid concentrations in follicular fluid of prepubertal goats and their effect on oocyte in vitro maturation and embryo development
Juvenile in vitro embryo transfer (JIVET) can increase the gain rate by reducing the generation interval, but the low quality of these oocytes leads to poor embryo production. In this study we hypothesised that oocyte quality could be related to omega 3 (linolenic acid, ALA) and omega 6 (linoleic acid, LA) concentrations in follicular fluid. No differences were found in ALA and LA concentrations in relation to follicle diameter. The addition of a high LA (omega 6) concentration to the IVM medium impaired oocyte competence to develop up to the blastocyst stage.
The zona pellucida (ZP) is an extracellular matrix formed by few glycoproteins that covers the oocyte and plays an important role during fertilisation. The identification of ZP glycoproteins in carnivores is relevant for an understanding of sperm–ZP interactions and species specificity. In this study, molecular and in silico analyses demonstrated the presence of the ZP1 gene in four families (Felidae, Mustelidae, Odobenidae and Ursidae), whereas in three families (Canidae, Otariidae and Phocidae) ZP1 is pseudogenised. This information could be relevant for the development of more specific and efficient contraceptive vaccines for carnivores.
RD16301Visfatin and resistin in gonadotroph cells: expression, regulation of LH secretion and signalling pathways
Resistin and visfatin, two adipokines, modulate reproduction in the gonads, but their actions at the hypothalamic–pituitary level are not known. We showed in vitro that these hormones decreased LH secretion from mouse gonadotroph cells and that in vivo visfatin may be a myokine as well as an adipokine in mice. Our findings suggest that these hormones may influence mouse female fertility by regulating LH secretion at the pituitary level.
RD17107Delayed onset of puberty in male offspring from bisphenol A-treated dams is followed by the modulation of gene expression in the hypothalamic–pituitary–testis axis in adulthood
The developing fetal brain is a potent target for endocrine chemical disruption by bisphenol A (BPA) because of its lipophilicity. In the present study, perinatal exposure to BPA changed the expression of genes regulating the hypothalamic–pituitary–testis (HPT) axis at the hypothalamic, pituitary and testes levels. Maternal exposure to BPA during fetal differentiation of the sexually dimorphic nucleus of the hypothalamus caused reprogramming of the HPT axis in adult male offspring
Juvenile in vitro embryo transfer (JIVET) can increase the genetic gain rate by reducing the generation interval, but the low quality of the oocytes and higher oxidative levels leads to poor embryo production. In the present study we hypothesised that melatonin could be a suitable tool for improving oocyte competence in juvenile goats. The results show that melatonin is present in the follicular fluid in juvenile goats and its addition to the in vitro maturation medium reduces oxidation and increases embryo development showing its potential role for JIVET.
RD16282Increased high molecular weight adiponectin, but decreased total adiponectin and kisspeptin, in central precocious puberty compared with aged-matched prepubertal girls
The factors initiating puberty have not been clearly revealed. The present study compared serum total and high molecular weight (HMW) adiponectin levels between girls with central precocious puberty (CPP) and age-matched controls. Total adiponectin levels were low, but HMW adiponectin levels were high in girls with CPP, which probably leads to decreased inhibition of gonadotrophin-releasing hormone neuronal activity. This may provide an insight into how adipokines are involved in the initiation of puberty.
RD16290Novel insights into the role of cell-free seminal mRNAs on semen quality and cryotolerance of spermatozoa in bulls (Bos taurus)
The selection of fertile males is important to augment fertility in cows, and breeding soundness examinations may not provide information on the fertility potential of bulls. Expression levels of seminal plasma RNAs may be used to predict the fertilising ability and cryotolerance of bull semen. Such non-invasive tests using seminal plasma may aid in the assessment of the health of the reproductive tract and in the selection of males for breeding.
RD17089Temperature gradients in vivo influence maturing male and female gametes in mammals: evidence from the cow
Is the temperature in a preovulatory follicle lower than in adjacent tissues? Using a novel non-invasive approach it was demonstrated that bovine follicular fluid before ovulation is cooler than the uterine surface and deep rectal tissues, whereas temperature is not reduced in follicles suffering ovulation failure. Follicular antral temperature could become a more valuable indicator of oocyte potential than a purely morphological assessment.
RD17136Effect of seminal plasma from high- and low-fertility bulls on cauda epididymal sperm function
Seminal plasma bathes the sperm upon ejaculation but its function has yet to be fully defined. This study sought to characterise the effect of seminal plasma from bulls of high or low fertility on sperm function. Seminal plasma reduced osmotic resistance of epididymal sperm, regardless of whether it was from high or low fertility bulls.
Enzymatic disaggregation of murine ovaries yielded higher numbers of morphologically intact follicules than previously reported. The molecular markers DAPI, CMXRos and alpha-tubulin were applied to assess post-harvest follicular morphology, mitochondrial function and cellular cytoskeleton respectively. A new preparation of animal origin-free collagenase IV caused more damage to follicular integrity, mitochondrial function and the cytoskeleton than enzyme-free mechanical isolation, but another preparation of collagenase IV ovarian disaggregation gave the highest yield of morphologically intact follicles containing viable mitochondria.
RD17111Splicing-related single nucleotide polymorphism of RAB, member of RAS oncogene family like 2B (RABL2B) jeopardises semen quality in Chinese Holstein bulls
Bull semen quality traits are vital parameters in evaluating the fertility potential of bulls. In the present study we demonstrated that a single nucleotide polymorphism within the exonic splicing enhancer motif caused aberrant splicing of the RAB, member of RAS oncogene family like 2B (RABL2B) gene, and that semen quality differed among Chinese Holstein bulls with different RABL2B genotypes. These results also provide a novel research idea for further elucidating the complex molecular mechanism underlying the regulation of bull fertility.
RD17100Mouse minipuberty coincides with gonocyte transformation into spermatogonial stem cells: a model for human minipuberty
Subsequent infertility and testicular cancer are major problems for boys with undescended testis. This study aims to examine whether mouse has similar postnatal hormonal changes as boys, so it can be used as a model to study human testis development. Our results demonstrated that mouse can be used as a model for human minipuberty.
The importance of the oviduct for different reproductive processes is known; however, the molecular mechanisms underlying oviductal regulation are not fully understood. The results of the present study demonstrate that gene expression in the oviduct is regulated during the oestrous cycle, with some oviductal proteins that could be related to several reproductive processes being described for the first time. These findings are very important in our understanding of the different processes that take place in the oviduct as fertilisation or early embryo development.
The expression of IL-11 is stimulated by LH/hCG in theca cells of preovulatory follicles via PKA-MAPK pathway and TLR4 activation. IL-11 increases progesterone production by stimulating StAR expression.
RD16386Effects of vitrification of cumulus-enclosed porcine oocytes at the germinal vesicle stage on cumulus expansion, nuclear progression and cytoplasmic maturation
Although oocyte cryopreservation is desired for the preservation of female germplasm, the developmental competence of vitrified porcine oocytes has been notoriously low due to unclear reasons. We investigated the effects of vitrification of porcine oocytes at the immature stage. Vitrification triggered premature nuclear resumption and nucleolar fragmentation from which some oocytes could recover, suggesting that enhancement of recovery processes might be a possible way to improve competency.
RD16366Development of pre-implantation porcine blastocysts cultured within alginate hydrogel systems either supplemented with secreted phosphoprotein 1 or conjugated with Arg-Gly-Asp Peptide
Deficiencies in the initiation of embryo elongation play a major role in early embryonic mortality in the pig. This study demonstrates that embryos encapsulated and cultured in alginate hydrogels with the integrin-binding Arg-Gly-Asp (RGD) sequence had increased embryonic survival and modulation of gene expression to support increased steroid production and adequate immune responsiveness, with a larger number exhibiting the morphological changes compared to other in vitro treatments in this study. These results illustrate the importance of the integrin-binding RGD peptide sequence for stimulating the initiation of embryo elongation and improving embryo survival.
RD16428Intracytoplasmic oxidative stress reverses epigenetic modifications in polycystic ovary syndrome
In PCOS, elevated levels of plasma androgens affects the quality of oocytes and ovulation rate, and causes anovulation in women of reproductive age. We studied the mechanism in a mouse model of PCOS, and showed that intracytoplasmic oxidative stress in oocyte via increase in histone acetylation and decrease in DNA methylation and histone dimethylation alters epigenetic modification. Consistently, mRNA expression of DNA methyltransferase-1 and histone deacetylase-1 was decreased with PCOS. There was a significant correlation between ROS production and increased histone acetylation. We concluded that antioxidant therapy in PCOS is a viable approach to reduce the adverse effects of epigenetic alterations.
RD17046Protective effects of polydatin on experimental testicular torsion and detorsion injury in rats
The notable pathophysiological event that occurs in testicular torsion–detorsion (T/D) is ischaemia–reperfusion (I/R), which lacks efficient clinical treatment. Herein we describe, for the first time, the protective effects of polydatin, a traditional Chinese medicine, against testicular T/D injury in the rat. Our findings may provide new clues for male infertility associated with testicular torsion.
RD16343Andrographolide disrupts meiotic maturation by blocking cytoskeletal reorganisation and decreases the fertilisation potential of mouse oocytes
Andrographolide should be prudently consumed by women attempting to conceive. The aim of the present study was to investigate the effects of andrographolide on oocytes. The findings indicate that andrographolide may disrupt mouse oocyte meiotic maturation and fertilisation potential by blocking cytoskeletal reorganisation. This study provides an important platform for understanding the potential adverse effects of andrographolide on female fertility.
RD16459Comparative analysis of granulosa cell gene expression in association with oocyte competence in FSH-stimulated Holstein cows
In cattle, a major limitation of the success of IVF is the developmental competence that oocytes acquire during ovarian stimulation. The aim of the present study was to better understand the follicular conditions associated higher versus lower embryo yields. We revealed that the level of follicular differentiation (timing) is the main cause of failure and provide new biomarkers to identify such follicular conditions to adapt the ovarian stimulation protocol to individual animals.
RD16455The egg coat zona pellucida 3 glycoprotein – evolution of its putative sperm-binding region in Old World murine rodents (Rodentia: Muridae)
Binding of spermatozoa to the egg coat is a key step in fertilisation and provides a mechanism for reproductive isolation and sexual selection. To test this theory we investigated the molecular evolution of the ZP3 gene and its encoded sperm-binding region in a diverse murine rodent group. Our analyses provides evidence for positive selection on the sperm-binding region in several murine lineages, but the nature of the selective force remains unresolved.
RD16442Supplementation of culture medium with L-carnitine improves the development and cryotolerance of in vitro-produced porcine embryos
Porcine embryos contain substantial amounts of lipid, with little known regarding its metabolic role during development. This study investigated the association of lipid metabolism with carbohydrate substrates in the culture medium and revealed positive effects of adding L-carnitine, a limiting co-factor required for lipolysis, on early cleavage and blastocyst quality. Supplementing medium with L-carnitine is a useful strategy to improve the success of porcine blastocyst vitrification.
RD16513Melatonin potentially acts directly on swine ovary by modulating granulosa cell function and angiogenesis
The reproductive effects of melatonin have been extensively studied, but its involvement in the local control of ovarian physiology has not yet been clarified. The present research, which was undertaken to study the effects of physiological levels of melatonin on the main parameters of swine granulosa cell function, documents that the hormone is involved in the modulation of ovarian follicle growth and development.
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.
Thyroid hormone receptors de novo transcription is essential for early bovine embryo development in vitro
Effect of exogenous progesterone on embryo size and ewe uterine gene expression in an ovine dam size model of maternal constraint
Changes in bone turnover and calcium homeostasis during pregnancy and lactation in mammals: A meta-analysis
Proteinaceous sperm motility inhibitory factor from the female Indian garden lizard, Calotes versicolor
Molecular cloning and epigenetic change detection of Kiss1 during seasonal reproduction in Chinese indigenous sheep
Seminal plasma and cryopreservation alter ram sperm surface carbohydrates and interactions with neutrophils
Effect of lemon essential oil (Citrus limon) and its major components (Limonene (R)-(+) and Limonene (S)-(-)) on cryopreservation of bull spermatozoid
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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Presence of vascular endothelial growth factor during the first half of IVM improves the meiotic and developmental competence of porcine oocytes from small folliclesReproduction, Fertility and Development 29 (10)Tra M. T. Bui, Khánh X. Nguyễn, Asako Karata, Pilar Ferré, Minh T. Trần, Takuya Wakai, Hiroaki Funahashi
Reproduction, Fertility and Development 29 (1)
Differential effects of high and low glucose concentrations during lipolysis-like conditions on bovine in vitro oocyte quality, metabolism and subsequent embryo developmentReproduction, Fertility and Development 29 (11)J. De Bie, W. F. A. Marei, V. Maillo, L. Jordaens, A. Gutierrez-Adan, P. E. J. Bols, J. L. M. R. Leroy
In vitro addition of docosahexaenoic acid improves the quality of cooled but not frozen–thawed stallion semenReproduction, Fertility and Development 29 (10)D. M. Silva, S. A. Holden, A. Lyons, J. C. Souza, S. Fair
Reproduction, Fertility and Development (Online Early)Anusha Singh, Puran Bora, Amitabh Krishna
Epigenetic changes of hepatic glucocorticoid receptor in sheep male offspring undernourished in uteroReproduction, Fertility and Development 29 (10)Stella Chadio, Basiliki Kotsampasi, Stylliani Taka, Emmanouil Liandris, Nikolaos Papadopoulos, Elias Plakokefalos
Low doses of bisphenol A can impair postnatal testicular development directly, without affecting hormonal or oxidative stress levelsReproduction, Fertility and Development 29 (11)Fernanda M. Ogo, Glaucia E. M. L. Siervo, Géssica D. Gonçalves, Rubens Cecchini, Flavia A. Guarnier, Janete Ap. Anselmo-Franci, Glaura S. A. Fernandes
Protein in culture and endogenous lipid interact with embryonic stages in vitro to alter calf birthweight after embryo vitrification and warmingReproduction, Fertility and Development 29 (10)E. Gómez, S. Carrocera, S. Uzbekova, D. Martín, A. Murillo, M. Alonso-Guervós, F. Goyache, M. Muñoz
Ontogeny of clock and KiSS-1 metastasis-suppressor (Kiss1) gene expression in the prepubertal mouse hypothalamusReproduction, Fertility and Development 29 (10)Cassandra C. Yap, Peter J. Mark, Brendan J. Waddell, Jeremy T. Smith
Gel-coated tubes extend above-freezing storage of honey bee (Apis mellifera) semen to 439 days with production of fertilised offspringReproduction, Fertility and Development 29 (10)Brandon K. Hopkins, Susan W. Cobey, Charles Herr, Walter S. Sheppard
Sperm gamma-aminobutyric acid type A receptor delta subunit (GABRD) and its interaction with purinergic P2X2 receptors in progesterone-induced acrosome reaction and male fertilityReproduction, Fertility and Development 29 (10)Wenming Xu, Ke Wang, Yan Chen, Xiao Tong Liang, Mei Kuen Yu, Huanxun Yue, M. Louise Tierney
Polydatin improves the developmental competence of bovine embryos in vitro via induction of sirtuin 1 (Sirt1)Reproduction, Fertility and Development 29 (10)Imran Khan, Sung Woo Kim, Kyung-Lim Lee, Seok-Hwan Song, Ayman Mesalam, M. M. R. Chowdhury, Zia Uddin, Ki Hun Park, Il-Keun Kong
Intrafollicular oestradiol production, expression of the LH receptor (LHR) gene and its isoforms, and early follicular deviation in Bos indicusReproduction, Fertility and Development 29 (10)S. Wohlres-Viana, E. K. N. Arashiro, M. A. Machado, L. S. A. Camargo, L. G. B. Siqueira, M. P. Palhao, J. H. M. Viana