Equine embryo research focuses on the early developmental stages of horses, encompassing the formation, growth, and differentiation of the embryo. This area of study is significant for understanding reproductive biology, improving breeding programs, and advancing assisted reproductive technologies in equines. Key aspects include the processes of fertilization, embryonic development, and implantation. Researchers investigate factors influencing embryonic viability, such as genetic and environmental influences, to enhance reproductive success rates. This page aggregates peer-reviewed research studies and scholarly articles that explore the biological mechanisms, technological advancements, and applied methodologies related to equine embryos.
Tsopp E, Kilk K, Gambini A, Tvrdà E, Viljaste-Seera A, Tuppits U, Jaakma Ü.Equine in vitro embryo production (IVP) via ovum pick-up (OPU) and intracytoplasmic sperm injection (ICSI) has gained prominence in sport-horse breeding due to high success rates, efficient semen utilization, and year-round embryo production, including from mares with fertility challenges or postmortem. However, assessing viability of early-stage ICSI-produced equine embryos remains challenging due to subjective morphological evaluations and limited visibility of developmental features. Non-invasive metabolomic analysis of culture media offers a promising alternative to enhance embryo selectio...
Cortez JV, Cervi D, Ruiz AJ, Grupen CG.Nicotinic acid (NA) treatment during in vitro maturation (IVM) has been shown to elevate nicotinamide adenine dinucleotide (NAD) levels and improve oocyte developmental competence. Suboptimal equine oocyte IVM systems currently limit the efficiency of viable embryo in vitro production. This study evaluated NA supplementation during IVM for cloned equine embryo production, using oocytes from abattoir-sourced ovaries and live mares via ovum pick-up (OPU). Abattoir-derived oocytes (n = 694) were treated without or with 50 or 200 μM NA during the 18 h holding period (Pre-IVM). Next, OPU-deriv...
Hedia M, Leroy JLMR, Loomans S, Benedetti C, Angel-Velez D, Chiers K, Govaere J, Van Soom A, Smits K.Lipopolysaccharide (LPS) in follicular fluid impairs steroid production and oocyte developmental competence in cows and mice. This study assessed LPS concentrations in equine follicular fluid and their association with steroid and some cytokine levels. Additionally, we evaluated whether LPS exposure during in vitro maturation (IVM) affects equine oocyte developmental competence. In experiment 1, follicular fluid from large follicles (>30 mm in diameter) was collected from 16 slaughterhouse mares, and concentrations of LPS, estradiol, progesterone, TNF-α, and IL-6 were measured. In experiment...
Martin-Pelaez S, de la Fuente A, Takahashi K, Monteiro H, Mendes M, Meyers S, Dini P.Despite significant advances, in vitro production (IVP) of equine embryos continues to lack standardised embryo classification criteria and is associated with increased rates of early pregnancy loss compared with in vivo-derived blastocysts. Objective: To evaluate morphokinetic characteristics of the first mitotic division and early embryonic development in IVP blastocysts and their association with embryo development, as well as pregnancy rate and early pregnancy loss following embryo transfer. Methods: Retrospective analysis of archived material and clinical records. Methods: We retrospectiv...
Segabinazzi L, Lisboa FP, Zutter B.Limited response to conventional therapies and rising antibiotic resistance emphasize the need for alternative treatments for infectious endometritis. A recent in vitro study showed promising antimicrobial activity of a commercial uterine cleaning sanitizer (UCS) against endometritis pathogens. However, its safety and efficacy in mares remain untested. Objective: This study aimed to assess the clinical safety of intrauterine infusion of UCS in reproductively sound mares. Methods: Exp. 1. Six healthy mares with endometrial biopsies classified as I or IIA had two intrauterine infusions of UCS pe...
Nemoto M, Kawanishi N, Kambayashi Y, Bannai H, Yamanaka T, Garvey M, Cullinane A, Yamayoshi S, Kawaoka Y, Tsujimura K.Reverse genetics (RG) technology is useful for quickly updating influenza vaccine strains. A high-yield backbone (i.e., six segments other than hemagglutinin and neuraminidase) derived from A/Puerto Rico/8/1934 (PR8) has been developed to improve the growth of avian and human influenza viruses. However, for equine influenza virus (EIV), an EIV-derived backbone may have better growth properties due to more-natural segment combinations. We compared the growth properties in eggs between the PR8 backbone and an EIV backbone from A/equine/Ibaraki/1/2007, a vaccine strain in Japan. The results showe...
Scarlet D, Schuler G, Malama E, Bollwein H, Bocci C, Colleoni S, Lazzari G, Galli C, Kowalewski MP.Despite recent improvements in equine ovum pick-up (OPU) combined with intracytoplasmic sperm injection (ICSI), there is still significant inter-individual variability. In this study, serum concentrations of the oocyte-secreted factors growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), as well as the GDF9/BMP15 complex, anti-Müllerian hormone (AMH), progesterone (P4), and free and conjugated estradiol-17β (E2), were determined in 59 mares (3-24 years) at the time of OPU. Mares were classified retrospectively, based on the number of embryos obtained, into low or...
Couto G, Grippo A, Ismer A, Hoogewijs M, Pedro B, Vasconcelos L, Santos G, Wilsher S.Vitrified in vitro-produced embryos can be successfully warmed in isotonic media at room temperature (RT; 22°C). However, this protocol has not been reported for in vivo embryos, which are more challenging to vitrify and warm. Study objectives were to see if vitrified in vivo embryos warmed in RT isotonic medium gave equivalent pregnancy rates to stepwise serial dilution warming, and if embryo size influenced the results. One hundred and seventeen embryos were divided into groups by size (G1:≤ 300 μm, n = 59; G2:> 300-400 μm, n = 33; G3:> 400-500 μm, n = ...
Wilsher S, Ismer A, Grippo A, Hoogewijs M, Bussade P, Kovacsy S.Different cryoprotectants can influence the ability of embryos to successfully survive vitrification and subsequent warming before transfer. Objective: To compare pregnancy rates for embryos ≤500 μm vitrified, without puncture or aspiration of the blastocoele cavity, with one of three commercial human embryo vitrification kits containing the same penetrating cryoprotectants (DMSO and EG) but varying in their non-penetrating cryoprotectants (NPCPAs; sucrose, trehalose, dextran serum supplement [DSS], and hydroxypropyl cellulose [HPC]). Methods: In vivo experiments. Methods: Embryos (n =â...
Barandalla M, Colleoni S, Perota A, Galli C, Lazzari G.The introduction of ovum pick-up, Intracytoplasmic Sperm Injection (ICSI) and in vitro embryo production in equine reproduction offers significant advantages, including overcoming infertility, high blastocyst yields, embryo cryopreservation, and preimplantation genetic testing (PGT) by collecting few cells extruded spontaneously during embryo growth using a mouth pipette. PGT enables embryo selection before transfer and is particularly useful in breeds with a preference for specific offspring genders, coat color, and detecting genetic diseases. Here, we present the first large-scale validation...
Fakhar-I-Adil M, Angel-Velez D, Araftpoor E, Amin QA, Hedia M, Bühler M, Gevaert K, Menten B, Van Soom A, Chuva de Sousa Lopes SM, Stoop D, De Roo C....In vitro maturation (IVM) of oocytes retrieved from ovum pick-up (OPU) or ovarian tissue (OT) is a standard approach for patients with specific conditions where prior hormonal stimulation is contraindicated. However, the developmental competence of oocytes matured in vitro is still inferior to that of oocytes matured in vivo. Capacitation IVM (CAPA-IVM) includes an extra step of pre-maturation culture (PMC) with c-type natriuretic peptide (CNP) as a meiotic arrestor to better synchronize cytoplasmic and nuclear maturity in oocytes by allowing the cytoplasm additional time to acquire essential ...
Hannan MA, Watanabe H, Takeyama A, Yoshida S, Wudamu D, Lkhagvasuren N, Claes A, Stout TAE, Cheong SH, Haneda S, Nambo Y.This study evaluated the viability of in vitro embryo production using ovum pick-up (OPU) and intracytoplasmic sperm injection (ICSI) as breeding techniques for pure and crossbred Hokkaido native ponies (n = 9). Oocytes were collected using transvaginal ultrasound-guided follicle aspiration. ICSI was performed on in vitro matured oocytes using frozen semen. Embryonic cultures were monitored using time-lapse cinematography. Blastocysts were cryopreserved and, after thawing, were transferred non-surgically into recipient mares. Over nine OPU sessions, the mean number of aspirated follicles was 2...
Jones CJP, Wilsher S, Aplin JD.We report a lectin histochemical study of oviductal ampulla and isthmus of thirteen mares euthanised at anestrus (1), estrus (5), 1-day post-ovulation (5) or diestrus (2). Staining with a panel of 5 lectins recognizing N-Acetylgalactosamine (GalNAc) residues revealed mosaicism in glycan expression in some specimens of ampulla and isthmus, peaking at estrus and 1-day post-ovulation (ov+1). In the ampulla, this presented as positively stained areas of both ciliated and non-ciliated cells in otherwise negatively-stained ampullae, ranging from small foci (diestrus) to areas up to 200 microns in le...
Kushida C, Usui T, Tamura N, Kasashima Y, Sato K, Arai K.This study evaluated the effects of cell adhesion substrates that lead to the generation of equine-induced pluripotent stem cells (eiPSC) from embryonic skin fibroblasts by lipofection of plasmid vectors expressing five reprogramming factors. The reprogramming efficiency of cells induced on the E8 fragment of laminin-511 (eiPSC-511) was higher than that on Geltrex containing laminin-111 as a major laminin (eiPSC-111), and supplementation with a cocktail of small molecular compounds increased the number of iPSC colonies on both substrates. In the cell proliferation assay, eiPSC-511 showed highe...
Cuervo-Arango J, Sala-Ayala L, Márquez-Moya A, MartÃnez-Bovà R.The objective of this study was to determine the effect of two aspiration pressures (75 vs. 150 mmHg), the follicle flushing method (injection pump controlled by a foot pedal vs. a plastic syringe) and the twisting of the OPU needle on oocyte recovery and in vitro embryo production. OPU data from a total of 104 warmblood sport mares belonging to a commercial OPU-ICSI program were collected as part of a prospective study split into three experiments. Each mare was used only once for OPU. In Experiment 1, the mares' follicles were aspirated using either a high aspiration pressure (flow rate of 1...
Felix MR, Dobbie T, Woodward E, Linardi R, Okada C, Santos R, Hinrichs K.We recently reported successful equine IVF using fresh semen pre-incubated for a prolonged period (22Â h) before co-culture with oocytes. In this study, we evaluated the feasibility of equine IVF with frozen-thawed sperm and evaluated capacitation-related changes in these sperm over the pre-incubation period. Sperm selected via a commercial sperm separation device (SSD) yielded significantly higher fertilization than did sperm selected by swim-up or by colloid centrifugation. Using the SSD method, fertilization rates with sperm pre-incubated for 15Â min, 3Â h, 6Â h, and 9Â h were 7.1, 22.2, 38...
Fedorka CE, Scoggin KE, Coleman SJ, Hatzel JN, Burleson MD, Troedsson MHT.Alterations during the early stages of embryo development have been associated with long-term effects on the fetus, neonate, and adult, but this has not been investigated in horses. In recent years, intracytoplasmic sperm injection (ICSI) has gained in commercial popularity in the equine population. Research suggests an association between ICSI-produced embryos and placental malformations, but there exists little understanding of the physiology involved. Therefore, we aim to produce a complete transcriptomic analysis of chorioallantois and provide potential pathways that may be impacted follow...
Ulaangerel T, Mu S, Sodyelalt J, Yi M, Zhao B, Hao A, Wen X, Han B, Bou G.The success or failure of embryo fixation is crucial for embryo attachment and later development. As an epithelial chorioallantoic placenta-type animal, the horse has a special process of embryo implantation, and the mechanism of embryo fixation in horses is still unclear. Methods: In this study, the structural and transcriptomic characteristics of endometrial tissue from the fixed and nonfixed sides of 20-day gestation embryos in Mongolian horses were investigated to search for important genes and potential molecular markers associated with the fixation phase of equine embryos. Results: A com...
Perera TRW, de Ruijter-Villani M, Gibb Z, Nixon B, Sheridan A, Stout TAE, Swegen A, Skerrett-Byrne DA.Embryo-maternal signaling during the establishment of pregnancy in horses remains one of the biggest mysteries in large animal physiology. Early pregnancy loss represents a major source of economic loss to the breeding industry. This study aimed to investigate the systemic changes associated with early pregnancy by mapping the proteome of blood plasma at 14 days in pregnant and non-pregnant mares. Methods: Plasma proteomes were analysed in commercially bred pregnant (n = 17) and non-pregnant (n = 17) Thoroughbred mares at 14 days after ovulation, using high-resolution mass spectrometry. D...
Jaworska J, Tobolski D, Salem SE, Kahler A, Wocławek-Potocka I, Mestre AM.Embryo implantation in the mare occurs just over one month after fertilization, coinciding with the production of chorionic gonadotropin. The factors that regulate this late implantation in the mare, and whether they are unique to horses or shared with more invasive embryo implantation in other species, remain poorly understood. This study aimed to determine and compare the transcriptome and subpopulations of endometrial cells before and after embryo implantation in the horse. Single-cell RNA sequencing was used to characterize the transcriptome of nearly 97,000 endometrial cells collected fro...
Arnold LC, Stout TAE, Beitsma MM, Deelen CC, Rizzo M, Leemans B, Galli C, Colleoni S, Lazzari G, Barandalla M, Claes ANJ.It is not known whether the likelihood of pregnancy after transfer of cryopreserved equine in vitro produced (IVP) blastocysts is related to post-thaw morphological characteristics, or could be impacted by other factors such as embryo biopsy prior to cryopreservation. This study examined associations between post-thaw morphological appearance of equine IVP embryos, and biopsy prior to slow-freezing, on the likelihood of pregnancy. Biopsy of IVP blastocysts was performed by harvesting cells herniated through the intra-cytoplasmic sperm injection (ICSI)-derived hole in the zona pellucida (ZP). M...
Sala-Ayala L, Cuervo-Arango J, Martinez-Bovi R.The case report discusses the implications of using different volumes of flushing media during embryo flushing (EF) procedures in a single donor mare that experienced multiple unintended pregnancies following negative EF outcomes. The study involved two sets of EF experiments over two years on a non-lactating Spanish purebred mare. The initial experiments (group 1, n=6) used 1L of flushing medium (Ringer´s Lactate), and subsequent tests (group 2, n=5) increased the volume to 2L, to assess whether a larger volume could enhance embryo recovery rates. After every EF procedure, no PGF2α was admi...
Broothaers K, Pascottini OB, Hedia M, Angel-Velez D, De Coster T, Peere S, Polfliet E, Van den Branden E, Govaere J, Van Soom A, Smits K.Previous studies in the horse highlight the potential benefit of prolonged in vitro maturation (IVM) (34Â h) compared to short IVM (24Â h) with or without prior oocyte holding, but little is known about the optimal IVM duration within this interval. To determine the effect of oocyte holding and duration of IVM ranged between 28 and 34Â h on nuclear maturation, cleavage, blastocyst formation, and pregnancy rates, a retrospective study was performed in an equine clinical OPU-ICSI setting. The study included data of 2114 aspirated oocytes from 201 OPU-ICSI sessions. Duration of IVM was divided in...
Du M, Li X, Bayinnamula , Wang N, Liu Y, Zhang L, Zhao Y, Dugarjaviin M.An important method for preserving equine germplasm is the cryopreservation of equine oocytes. Due to its ease, rapidity and affordability, vitrification freezing has taken over as the primary method of horse oocyte cryopreservation. The vitrification cryoprotectants utilized in this investigation were Ethylene glycol (E), Dimethyl sulfoxide (D), Sucrose (S), and Ficoll (F). According to the oocyte volume alteration, the treatment time was 39 s in equilibrium solution ED10 (10 % EG + 10 % DMSO), 32 s in equilibrium solution ED15 (15 % EG + 15 % DMSO), while 20 s in equilibrium so...
Kehlbeck A, Blanco M, Venner M, Freise F, Gunreben B, Sieme H.Fragile foal syndrome (FFS) or warmblood FFS Type 1 (WFFS) is a recessive, autosomal, hereditary, genetic defect causing late abortions, stillbirths and non-viable foals. Whether early pregnancy losses occur is unknown. Objective: To investigate how WFFS affects pregnancies and whether early pregnancy losses occur in WFFS matings and if there is a difference in pregnancy success between matings where both parents were allele carriers and those where only mare or stallion were WFFS carriers. Methods: Retrospective cohort study. Methods: Breeding records from a Warmblood stud farm were evaluated...
GabryÅ› J, Pietras N, Kowal-Mierzwa W, Karnas E, Andronowska A, Nowak A, Kochan J, Bugno-Poniewierska M.The efficacy of in vitro embryo production (IVEP) in equines is relatively limited compared to other species due to the lack of a reliable superovulation technique, limited availability of cumulus oocyte complexes (COCs), low in vitro oocyte maturation (IVM) and fertilization rates. Extracellular vesicles (EVs), which are nanoparticles involved in intercellular signaling in the ovarian environment, have shown potential as supplements to improve oocyte development during IVM. This study tested the hypothesis that EVs from small (< 20Â mm) ovarian follicles could enhance fertilization rates in m...
Kenney RM.Normal histologic changes that occurred in concert with the ovarian cycle were evaluated. When combined with behavioral and physical findings, this evaluation enabled determination when there was synchrony between ovarian steroids (estrogen and progesterone) and their target tissues of endometrium. A system for classification as well as definitions of patterns types and degrees of severity of inflammatory and fibrotic changes was developed to facilitate a uniform system for description and communication. An attempt was made to relate the pathologic changes to their effect on fertility when fer...
Ezashi T, Yuan Y, Roberts RM.This review deals with the latest advances in the study of embryonic stem cells (ESC) and induced pluripotent stem cells (iPSC) from domesticated species, with a focus on pigs, cattle, sheep, goats, horses, cats, and dogs. Whereas the derivation of fully pluripotent ESC from these species has proved slow, reprogramming of somatic cells to iPSC has been more straightforward. However, most of these iPSC depend on the continued expression of the introduced transgenes, a major drawback to their utility. The persistent failure in generating ESC and the dependency of iPSC on ectopic genes probably s...
Klein C, Scoggin KE, Ealy AD, Troedsson MH.Establishment and maintenance of pregnancy are critically dependent on embryo-maternal communication during the preimplantation period. To gain new insights into this complex process in the horse, transcriptional profiling of Day 13.5 pregnant and cyclic endometrial tissue samples was carried out using custom-designed microarrays. Selected array data were validated using quantitative RT-PCR, and proteins of interest were localized using immunohistochemistry. One hundred and six transcripts were up-regulated, whereas 47 transcripts showed lower expression levels in pregnant mares, that is, were...
Muller B, Lewis N, Adeniyi T, Leese HJ, Brison DR, Sturmey RG.Mitochondria provide the major source of ATP for mammalian oocyte maturation and early embryo development. Oxygen Consumption Rate (OCR) is an established measure of mitochondrial function. OCR by mammalian oocytes and embryos has generally been restricted to overall uptake and detailed understanding of the components of OCR dedicated to specific molecular events remains lacking. Here, extracellular flux analysis (EFA) was applied to small groups of bovine, equine, mouse and human oocytes and bovine early embryos to measure OCR and its components. Using EFA, we report the changes in mitochondr...
Leemans B, Gadella BM, Stout TA, De Schauwer C, Nelis H, Hoogewijs M, Van Soom A.In contrast to man and many other mammalian species, conventional in vitro fertilization (IVF) with horse gametes is not reliably successful. The apparent inability of stallion spermatozoa to penetrate the zona pellucida in vitro is most likely due to incomplete activation of spermatozoa (capacitation) because of inadequate capacitating or fertilizing media. In vivo, the oviduct and its secretions provide a microenvironment that does reliably support and regulate interaction between the gametes. This review focuses on equine sperm-oviduct interaction. Equine sperm-oviduct binding appears to be...
Choi YH, Varner DD, Love CC, Hartman DL, Hinrichs K.Work with lyophilized sperm helps delineate the factors required for successful fertilization. We investigated the use of lyophilized sperm in equine embryo production. In Experiment 1, sperm DNA fragmentation index was not affected by three freeze/thaw or lyophilization cycles. In Experiment 2, oocytes injected with lyophilized sperm or with sperm from a treatment in which lyophilized sperm were suspended in sperm cytoplasmic extract (SE) yielded blastocyst development rates of 0 and 28% respectively (P < 0.05). In Experiment 3, blastocyst development rate was 6-11% after injection of sper...
Iqbal K, Chitwood JL, Meyers-Brown GA, Roser JF, Ross PJ.Formation of the inner cell mass (ICM) and trophectoderm (TE) marks the first differentiation event in mammalian development. These two cell types have completely divergent fates for the remainder of the developmental process. The molecular mechanisms that regulate ICM and TE formation are poorly characterized in horses. The objective of this study was to establish the transcriptome profiles of ICM and TE cells from horse blastocysts using RNA sequencing (RNA-seq). A total of 12 270 genes were found to be expressed in either lineage. Global analysis of the transcriptome profiles by unsupervi...
Galli C, Colleoni S, Duchi R, Lagutina I, Lazzari G.Development of assisted reproductive technologies in horses has been relatively slow compared to other domestic species, namely ruminants and pigs. The scarce availability of abattoir ovaries and the lack of interest from horse breeders and breed associations have been the main reasons for this delay. Progressively though, the technology of oocyte maturation in vitro has been established followed by the application of ICSI to achieve fertilization in vitro. Embryo culture was initially performed in vivo, in the mare oviduct or in the surrogate sheep oviduct, to achieve the highest embryo devel...
Hinrichs K.A wide variety of assisted reproductive techniques (ARTs) are available to aid in managing aspects of equine reproduction. Embryo recovery and transfer can be used to obtain more than one foal per mare per year, and to obtain foals from mares that cannot carry a foal to term. Oocyte recovery and either transfer to the oviduct of an inseminated recipient mare (oocyte transfer), or intracytoplasmic sperm injection (ICSI) and embryo culture can be used to obtain foals from mares with some types of subfertility, such as problems of the tubular tract. ICSI can be used to obtain foals when sperm num...
Li X, Zhou SG, Imreh MP, Ahrlund-Richter L, Allen WR.Inner cell mass (ICM) cells were isolated immunosurgically from day 7-8 horse blastocysts and, after proliferation in vitro for 15-28 passages, three lines of cells were confirmed to be embryonic stem (ES) cells by their continued expression of alkaline phosphatase activity and their ability to bind antisera specific for the recognized stem cell markers, SSEA-1, TRA-1-60, TRA-1-81, and the key embryonic gene Oct-4. When maintained under feeder cell-free conditions in vitro, the three lines of cells differentiated into cells of ectodermal, endodermal, and mesodermal lineages. However, they did ...
Betteridge KJ, Eaglesome MD, Mitchell D, Flood PF, Beriault R.Forty nine embryos, twenty unfertilized eggs and five other fresh eggs of 'doubtful' status have been recovered from 58 pony mares in 122 flushes up to 22 days after ovulation. The fresh egg or embryo recovery rate was 78% with surgical methods (or at slaughter) and 40-60% with non-surgical methods of recovery. The fertilization rate was about 70%. It has been confirmed that horse embryos normally enter the uterus as blastocysts 5-6 days after ovulation. Three features of early embryo morphology have become clearer upon comparison with unfertilized eggs of similar ages; early embryos are often...
Galli C, Crotti G, Notari C, Turini P, Duchi R, Lazzari G.Embryo production by in vitro techniques has increased steadily over the years. For cattle where this technology is more advanced and is applied more, the number of in vitro produced embryos transferred to final recipients was over 30,000 in 1998. An increasing proportion of in vitro produced embryos are coming from oocytes collected from live donors by ultrasound-guided follicular aspiration (ovum pick up, OPU). This procedure allows the repeated production of embryos from live donors of particular value and is a serious alternative to superovulation. Ovum pick up is a very flexible technique...
Raudsepp T, Chowdhary BP.The association between chromosomal abnormalities and reduced fertility in domestic animals is well recorded and has been studied for decades. Chromosome aberrations directly affect meiosis, gametogenesis, and the viability of zygotes and embryos. In some instances, balanced structural rearrangements can be transmitted, causing fertility problems in subsequent generations. Here, we aim to give a comprehensive overview of the current status and future prospects of clinical cytogenetics of animal reproduction by focusing on the advances in molecular cytogenetics during the genomics era. We descr...
Smits K, Willems S, Van Steendam K, Van De Velde M, De Lange V, Ververs C, Roels K, Govaere J, Van Nieuwerburgh F, Peelman L, Deforce D, Van Soom A.During maternal recognition of pregnancy (MRP), a conceptus-derived signal leads to the persistence of the corpus luteum and the maintenance of gestation. In the horse, the nature of this signal remains to be elucidated. Several studies have focused on the changes in gene expression during MRP, but little information exists at the protein level. The aim of this study was to identify the proteins at the embryo-maternal interface around signalling of MRP in the horse (day 13) by means of mass spectrometry. A distinct influence of pregnancy was established, with 119 proteins differentially expres...
Klein C, Troedsson MH.Establishment and maintenance of pregnancy are critically dependent on embryo-maternal communication during the preimplantation period. The horse is one of the few domestic species in which the conceptus-derived pregnancy recognition signal has not been identified. To gain new insights into the factors released by the equine conceptus, transcriptional profiling analyses of conceptuses retrieved 8, 10, 12, and 14 days after ovulation were performed using a whole-genome microarray. Selected array data were confirmed using quantitative PCR, and the expression of proteins of interest was confirmed...
Carnevale EM, Ginther OJ.The uterine function and reproductive efficiency of 31 nonlactating pony mares were compared for two age groups: young (5 to 7 years, n=9) and old (>/=15 years, n=22). For pregnant mares, differences between age groups were not significant for the diameter of the largest follicle, cross-sectional area of the corpus luteum, growth profile of the embryonic vesicle or embryo mobility characteristics. Uterine contractility scores were lower (P<0.05), day of fixation of the embryonic vesicle was later (P<0.05), and uterine tone tended (P<0.10) to be lower in the old than the young mares...
Suire S, Stewart F, Beauchamp J, Kennedy MW.The equine conceptus is surrounded by a fibrous capsule that persists until about day 20 of pregnancy, whereupon the capsule is lost, the conceptus attaches to the endometrium and placentation proceeds. Before attachment, the endometrium secretes in abundance a protein of the lipocalin family, uterocalin. The cessation of secretion coincides with the end of the period during which the conceptus is enclosed in its capsule, suggesting that uterocalin is essential for the support of the embryo before direct contact between maternal and foetal tissues is established. Using recombinant protein and ...
Guest DJ, Allen WR.Embryonic stem-like (ES-like) cells have now been derived from the inner cell mass (ICM) of horse embryos at the blastocyst stage. Because they have been shown to express cell-surface antigens found in both human and mouse ES cells, the present study investigated gene expression patterns in day-7 horse blastocysts from which the horse ES-like cells had been derived originally. The genes studied included Oct-4, stage-specific embryonic antigen-1 (SSEA-1), SSEA-3, SSEA-4, tumor rejection antigen-1-60 (TRA-1-60), TRA-1-81, and alkaline phosphatase activity, and whereas all three of the SSEA antig...
Ambruosi B, Lacalandra GM, Iorga AI, De Santis T, Mugnier S, Matarrese R, Goudet G, Dell'aquila ME.Lipid droplets (LDs) and mitochondria in the ooplasm are essential for energy production required for maturation, fertilization and embryo development. This study investigates the correlations between cytoplasmic LDs polar aggregation and: (1) nuclear maturation (Experiment 1); (2) mitochondrial (mt) distribution pattern and localization (Experiment 2); (3) fertilization and embryonic development after intracytoplasmic sperm injection (ICSI; Experiment 3) in equine oocytes recovered from slaughtered mares and matured in vitro. Morphologically normal oocytes were selected after culture and cate...
Bauersachs S, Wolf E.Different reproductive strategies evolved in various mammalian groups to achieve recognition, establishment and maintenance of pregnancy. The complexity of these processes is reflected by a high incidence of embryonic loss during this critical period in many mammalian species. Besides studies in mice and humans a number of transcriptome studies of endometrial tissue samples and also of early embryos have been performed during the pre-implantation phase in cattle, swine and horse to identify genes associated with embryo-maternal interaction. Results of these studies are reviewed and compared be...
Salamone DF, Canel NG, RodrÃguez MB.Intracytoplasmic sperm injection (ICSI) has become a useful technique for clinical applications in the horse-breeding industry. However, both ICSI blastocyst and offspring production continues to be limited for most farm and wild species. This article reviews technical differences of ICSI performance among species, possible biological and methodological reasons for the variable efficiency and potential strategies to improve the outcomes. One of the major applications of ICSI in animal production is the reproduction of high-value specimens. Unfortunately, some domestic species like the bovine s...
Lagutina I, Lazzari G, Duchi R, Colleoni S, Ponderato N, Turini P, Crotti G, Galli C.The objective of the present work was to investigate and clarify the factors affecting the efficiency of somatic cell nuclear transfer (NT) in the horse, including embryo reconstruction, in vitro culture to the blastocyst stage, embryo transfer, pregnancy monitoring and production of offspring. Matured oocytes, with zona pellucida or after zona removal, were fused to cumulus cells, granulosa cells, and fetal and adult fibroblasts, and fused couplets were cultured in vitro. Blastocyst development to Day 8 varied significantly among donor cells (from 1.3% to 16%, P < 0.05). In total, 137 nuclear...
Swegen A, Grupen CG, Gibb Z, Baker MA, de Ruijter-Villani M, Smith ND, Stout TAE, Aitken RJ.Early pregnancy in the mare is a poorly understood, high risk period during which the embryo communicates its presence to the maternal endometrium. Remarkably, the maternal recognition of pregnancy signal is unknown in the horse. This study aimed to profile the proteins secreted by equine blastocysts into their immediate environment, along with proteins contained in the blastocoel and within the acellular embryo capsule. Embryos were recovered on day 8 after ovulation and cultured for 48Â hours. Secretomes of day 9 and day 10 embryos were analyzed by LC-MS/MS and supported by analysis of blast...
Ginther OJ.Gray-scale ultrasonic imaging (UI) was introduced in 1980 and initially was used to examine clinically the reproductive tract of mares. By 1983 in mares and 1984 in heifers/cows, UI had become a tool for basic research. In each species, transrectal gray-scale UI has been used extensively to characterize follicle dynamics and investigate the gonadotropic control and hormonal role of the follicles. However, the use of transrectal UI has also disclosed and characterized many other aspects of reproduction in each species, including (1) endometrial echotexture as a biological indicator of circulati...
Smits K, De Coninck DI, Van Nieuwerburgh F, Govaere J, Van Poucke M, Peelman L, Deforce D, Van Soom A.Although the equine oviduct clearly affects early embryo development and the selective transport of equine embryos through the oviduct indicates a reciprocal interaction, the influence of the embryo on gene expression in the oviduct remains to be determined in the horse. The aim of this study was to examine this by means of RNA sequencing. Four days after ovulation, epithelial cells ipsilateral and contralateral to the ovulation side from five cyclic and five pregnant mares were collected from the oviduct. RNA was extracted, samples were sequenced, and data analysis was performed to determine ...
Gonzales DS, Jones JM, Pinyopummintr T, Carnevale EM, Ginther OJ, Shapiro SS, Bavister BD.The behaviour of bovine, equine and human blastocysts was studied in vitro by time-lapse videomicrography and computer imaging. This study revealed that cytoplasmic extensions of the trophectoderm ['trophectoderm projections' (TEP)] were expressed by embryos of all three species, prior to or during zona escape. Bovine and human blastocysts escaped their zonae with a combination of blastocoele expansion, collapse and re-expansion coupled with the penetration of the zona pellucida by TEP. In equine embryos, after several cycles of blastocoele expansion and collapse, trophectoderm ruptured the zo...
