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Topic:Developmental Biology

Developmental biology in horses involves the study of the processes by which horses grow and develop from a single fertilized egg into a fully formed organism. This field encompasses various stages, including embryonic development, fetal growth, and postnatal maturation. Researchers in this area examine cellular differentiation, gene expression, and morphogenetic movements that contribute to the formation of tissues and organs in equine species. Key topics include the molecular mechanisms that regulate developmental pathways, the influence of genetic and environmental factors on development, and the identification of developmental disorders. This page compiles peer-reviewed research studies and scholarly articles that explore the fundamental aspects of equine developmental biology, offering insights into the mechanisms driving normal and abnormal development in horses.
Development of novel gene carrier using modified nano hydroxyapatite derived from equine bone for osteogenic differentiation of dental pulp stem cells.
Bioactive materials    February 13, 2021   Volume 6, Issue 9 2742-2751 doi: 10.1016/j.bioactmat.2021.01.020
Lee MC, Seonwoo H, Jang KJ, Pandey S, Lim J, Park S, Kim JE, Choung YH, Garg P, Chung JH.Hydroxyapatite (HA) is a representative substance that induces bone regeneration. Our research team extracted nanohydroxyapatite (EH) from natural resources, especially equine bones, and developed it as a molecular biological tool. Polyethylenimine (PEI) was used to coat the EH to develop a gene carrier. To verify that PEI is well coated in the EH, we first observed the morphology and dispersity of PEI-coated EH (pEH) by electron microscopy. The pEH particles were well distributed, while only the EH particles were not distributed and aggregated. Then, the existence of nitrogen elements of PEI ...
Equine maternal aging affects oocyte lipid content, metabolic function and developmental potential.
Reproduction (Cambridge, England)    February 5, 2021   Volume 161, Issue 4 399-409 doi: 10.1530/REP-20-0494
Catandi GD, Obeidat YM, Broeckling CD, Chen TW, Chicco AJ, Carnevale EM.Advanced maternal age is associated with a decline in fertility and oocyte quality. We used novel metabolic microsensors to assess effects of mare age on single oocyte and embryo metabolic function, which has not yet been similarly investigated in mammalian species. We hypothesized that equine maternal aging affects the metabolic function of oocytes and in vitro-produced early embryos, oocyte mitochondrial DNA (mtDNA) copy number, and relative abundance of metabolites involved in energy metabolism in oocytes and cumulus cells. Samples were collected from preovulatory follicles from young (≤1...
Pathways involved in pony body size development.
BMC genomics    January 18, 2021   Volume 22, Issue 1 58 doi: 10.1186/s12864-020-07323-1
Fang J, Zhang D, Cao JW, Zhang L, Liu CX, Xing YP, Wang F, Xu HY, Wang SC, Ling Y, Wang W, Zhang YR, Zhou HM.The mechanism of body growth in mammals is poorly understood. Here, we investigated the regulatory networks involved in body growth through transcriptomic analysis of pituitary and epiphyseal tissues of smaller sized Debao ponies and Mongolian horses at the juvenile and adult stages. Results: We found that growth hormone receptor (GHR) was expressed at low levels in long bones, although growth hormone (GH) was highly expressed in Debao ponies compared with Mongolian horses. Moreover, significant downregulated of the GHR pathway components m-RAS and ATF3 was found in juvenile ponies, which slow...
Gross Motor Skills and Gait Performance in Two- and Three-year-old Children With Developmental Delay Participating in Hippotherapy.
Journal of equine veterinary science    December 30, 2020   Volume 99 103359 doi: 10.1016/j.jevs.2020.103359
Brady HA, James CR, Dendy DW, Irwin TA, Thompson LD, Camp TM.This study examined the effects of 15 sessions of hippotherapy (HPOT) on gross motor skills in children (aged 2-3 years) with gross motor developmental delay (DD) (n = 11) in comparison with age-based controls without DD (n = 6). Gross motor skills in both groups were assessed with the Battelle Developmental Inventory 2nd Edition, and gait parameters were measured using a computerized gait analysis system prestudy and poststudy. The DD group took part in 15 sessions of HPOT, and the control (CON) group did not participate in any equine activities. The statistical analysis examined preinterv...
A Review of OCT4 Functions and Applications to Equine Embryos.
Journal of equine veterinary science    December 24, 2020   Volume 98 103364 doi: 10.1016/j.jevs.2020.103364
Hisey E, Ross PJ, Meyers SA.OCT4 is a core transcription factor involved in pluripotency maintenance in the early mammalian embryo. The POU5F1 gene that encodes the OCT4 protein is highly conserved across species, suggesting conserved function. However, studies in several species including mice, cattle, and pigs, suggest that there are differences in where and when OCT4 is expressed. Specifically, in the horse, several studies have shown that exposure to the uterine environment may be necessary to induce OCT4 expression restriction to the inner cell mass (ICM) of the developing embryo, suggesting that there may be equine...
Development and Pathology of the Equine Mammary Gland.
Journal of mammary gland biology and neoplasia    December 5, 2020   Volume 26, Issue 2 121-134 doi: 10.1007/s10911-020-09471-2
Hughes K.An understanding of the anatomy, histology, and development of the equine mammary gland underpins study of the pathology of diseases including galactorrhoea, agalactia, mastitis, and mammary tumour development. This review examines the prenatal development of the equine mammary gland and the striking degree to which the tissue undergoes postnatal development associated with the reproductive cycle. The gland is characterised by epithelial structures arranged in terminal duct lobular units, similar to those of the human breast, supported by distinct zones of intra- and interlobular collagenous s...
Derivation of Intermediate Pluripotent Stem Cells Amenable to Primordial Germ Cell Specification.
Cell stem cell    December 2, 2020   Volume 28, Issue 3 550-567.e12 doi: 10.1016/j.stem.2020.11.003
Yu L, Wei Y, Sun HX, Mahdi AK, Pinzon Arteaga CA, Sakurai M, Schmitz DA, Zheng C, Ballard ED, Li J, Tanaka N, Kohara A, Okamura D, Mutto AA, Gu Y....Dynamic pluripotent stem cell (PSC) states are in vitro adaptations of pluripotency continuum in vivo. Previous studies have generated a number of PSCs with distinct properties. To date, however, no known PSCs have demonstrated dual competency for chimera formation and direct responsiveness to primordial germ cell (PGC) specification, a unique functional feature of formative pluripotency. Here, by modulating fibroblast growth factor (FGF), transforming growth factor β (TGF-β), and WNT pathways, we derived PSCs from mice, horses, and humans (designated as XPSCs) that are permissive for dire...
Kinetics of placenta-specific 8 (PLAC8) in equine placenta during pregnancy and placentitis.
Theriogenology    November 5, 2020   Volume 160 81-89 doi: 10.1016/j.theriogenology.2020.10.041
El-Sheikh Ali H, Scoggin K, Linhares Boakari Y, Dini P, Loux S, Fedorka C, Esteller-Vico A, Ball B.Placenta-specific 8 (PLAC8) is one of the placenta-regulatory genes which is highly conserved among eutherian mammals. However, little is known about its expression in equine placenta (chorioallantois; CA and endometrium; EN) during normal and abnormal pregnancy. Therefore, the current study was designed to 1) elucidate the expression of PLAC8 in equine embryonic membranes during the preimplantation period, 2) characterize the expression profile of PLAC8 in equine CA (45d, 4mo, 6mo, 10 mo, 11 mo and postpartum) and EN (14d, 4mo, 6mo, 10 mo, and 11 mo) obtained from pregnant mares (n = 4/...
Allometric growth in mass by the brain of mammals.
Anatomical record (Hoboken, N.J. : 2007)    November 5, 2020   Volume 304, Issue 7 1551-1561 doi: 10.1002/ar.24555
Packard GC.I re-examined published data for ontogenetic change in relative mass of the brain in six species of mammal (i.e., sheep, pig, cow, horse, rat, cat) to illustrate an insidious problem with conventional analyses of brain-body allometry. Graphical displays of logarithmic transformations of the original data for each species give the appearance of two discrete mathematical distributions, but untransformed observations nonetheless conform to a single distribution that is well described by a single, nonlinear equation. The concept of biphasic, allometric growth by the brain consequently is an artifa...
A high glucose concentration during early stages of in vitro equine embryo development alters expression of genes involved in glucose metabolism.
Equine veterinary journal    October 12, 2020   Volume 53, Issue 4 787-795 doi: 10.1111/evj.13342
Sánchez-Calabuig MJ, Fernández-González R, Hamdi M, Smits K, López-Cardona AP, Serres C, Macías-García B, Gutiérrez-Adán A.Equine embryos exhibit an unusual pattern of glucose tolerance in vitro and are currently cultured in hyperglycaemic conditions. Objective: Our main objective was to analyse the effect of different glucose concentrations on in vitro-produced equine embryo development and quality. Methods: Experiments comparing in vitro and in vivo produced embryos. Methods: Oocytes (n = 641) were collected from post-mortem ovaries, matured in vitro and fertilised by intracytoplasmic sperm injection (ICSI). Embryo culture was divided from Day 0 to Day 4 and from Day 4 to Day 9 in three groups: 5-10 (5 and 10...
Horse ooplasm supports in vitro preimplantation development of zebra ICSI and SCNT embryos without compromising YAP1 and SOX2 expression pattern.
PloS one    September 11, 2020   Volume 15, Issue 9 e0238948 doi: 10.1371/journal.pone.0238948
Gambini A, Duque Rodríguez M, Rodríguez MB, Briski O, Flores Bragulat AP, Demergassi N, Losinno L, Salamone DF.Several equids have gone extinct and many extant equids are currently considered vulnerable to critically endangered. This work aimed to evaluate whether domestic horse oocytes support preimplantation development of zebra embryos obtained by intracytoplasmic sperm injection (ICSI, zebroid) and cloning, and to study the Hippo signaling pathway during the lineage specification of trophectoderm cells and inner cell mass cells. We first showed that zebra and horse sperm cells induce porcine oocyte activation and recruit maternal SMARCA4 during pronuclear formation. SMARCA4 recruitment showed to be...
The transcription factor scleraxis differentially regulates gene expression in tenocytes isolated at different developmental stages.
Mechanisms of development    August 11, 2020   Volume 163 103635 doi: 10.1016/j.mod.2020.103635
Paterson YZ, Evans N, Kan S, Cribbs A, Henson FMD, Guest DJ.The transcription factor scleraxis (SCX) is expressed throughout tendon development and plays a key role in directing tendon wound healing. However, little is known regarding its role in fetal or young postnatal tendons, stages in development that are known for their enhanced regenerative capabilities. Here we used RNA-sequencing to compare the transcriptome of adult and fetal tenocytes following SCX knockdown. SCX knockdown had a larger effect on gene expression in fetal tenocytes, affecting 477 genes in comparison to the 183 genes affected in adult tenocytes, indicating that scleraxis-depend...
Generation of induced pluripotent stem cells from large domestic animals.
Stem cell research & therapy    June 25, 2020   Volume 11, Issue 1 247 doi: 10.1186/s13287-020-01716-5
Bressan FF, Bassanezze V, de Figueiredo Pessôa LV, Sacramento CB, Malta TM, Kashima S, Fantinato Neto P, Strefezzi RF, Pieri NCG, Krieger JE....Induced pluripotent stem cells (iPSCs) have enormous potential in developmental biology studies and in cellular therapies. Although extensively studied and characterized in human and murine models, iPSCs from animals other than mice lack reproducible results. Herein, we describe the generation of robust iPSCs from equine and bovine cells through lentiviral transduction of murine or human transcription factors Oct4, Sox2, Klf4, and c-Myc and from human and murine cells using similar protocols, even when different supplementations were used. The iPSCs were analyzed regarding morphology, gene and...
Equine endometrial development during late fetal and postnatal periods.
Theriogenology    June 10, 2020   Volume 156 155-161 doi: 10.1016/j.theriogenology.2020.06.006
Fraser NS, Wilborn RR, Johnson AK, Braden TD, Wiley AA, Canisso IF, Bartol FF.Equine uterine development, including endometrial histogenesis, begins prenatally and is completed postnatally. Little is known about this process in the horse. Uterine tissue was acquired from 38 foals, ranging in developmental age from gestational day (GD) 300 to postnatal day (PND) 180, for assessment of endometrial histogenesis. Patterns of endometrial cell proliferation were evaluated by multispectral imaging of uterine tissue sections stained immunofluorescently for Ki-67. Labeling index (LI, % labeled cells) for Ki-67 was calculated for each endometrial cell compartment (luminal epithel...
Morphological Characteristics of the Placenta and Umbilical Cord of Arabian Mares Foaling in the United Arab Emirates.
Journal of equine veterinary science    May 28, 2020   Volume 91 103124 doi: 10.1016/j.jevs.2020.103124
Wilsher S, Bowker A, Silva J, Allen WRT.A total of 127 normal placentas from Arabian mares resident in the United Arab Emirates were examined. The mean linear dimensions of the placenta were, on average, 84% of those previously recorded for the placentas of the Thoroughbred. Significant differences in the size of the allantochorion between primigravid and multiparous mares were seen only in the linear dimensions of the body portion. The pregnant horn was more commonly on the right than left side of the uterus (P = .01; 74/127; 58%). Cord attachment was primarily at the base of the two placental horns (112/127; 88%), with the remain...
Genome-wide transcriptome analysis reveals equine embryonic stem cell-derived tenocytes resemble fetal, not adult tenocytes.
Stem cell research & therapy    May 19, 2020   Volume 11, Issue 1 184 doi: 10.1186/s13287-020-01692-w
Paterson YZ, Cribbs A, Espenel M, Smith EJ, Henson FMD, Guest DJ.Tendon injuries occur frequently in human and equine athletes. Treatment options are limited, and the prognosis is often poor with functionally deficient scar tissue resulting. Fetal tendon injuries in contrast are capable of healing without forming scar tissue. Embryonic stem cells (ESCs) may provide a potential cellular therapeutic to improve adult tendon regeneration; however, whether they can mimic the properties of fetal tenocytes is unknown. To this end, understanding the unique expression profile of normal adult and fetal tenocytes is crucial to allow validation of ESC-derived tenocytes...
Normal regression of the internal umbilical remnant structures in Standardbred foals.
Equine veterinary journal    April 30, 2020   Volume 52, Issue 6 876-883 doi: 10.1111/evj.13267
McCoy AM, Lopp CT, Kooy S, Migliorisi AC, Austin SM, Wilkins PA.Diseases affecting the internal umbilical remnant are common in foals. Ultrasound is used to diagnose abnormalities of these structures, and to monitor treatment. However, little is known about the progression of normal internal umbilical remnant regression. Objective: To document normal regression of the umbilical remnant in foals over the first 6 weeks of life. Methods: Prospective, longitudinal cohort study. Methods: Weekly ultrasound examinations were performed beginning at 1 day of age in 34 healthy Standardbred foals. The umbilical vein was measured in cross section just cranial to the...
Transcriptome profiling of developing testes and spermatogenesis in the Mongolian horse.
BMC genetics    April 28, 2020   Volume 21, Issue 1 46 doi: 10.1186/s12863-020-00843-5
Li B, He X, Zhao Y, Bai D, Du M, Song L, Liu Z, Yin Z, Manglai D.Horse testis development and spermatogenesis are complex physiological processes. To study these processes, three immature and three mature testes were collected from the Mongolian horse, and six libraries were established using high-throughput RNA sequencing technology (RNA-Seq) to screen for genes related to testis development and spermatogenesis. A total of 16,237 upregulated genes and 8,641 downregulated genes were detected in the testis of the Mongolian horse. These genes play important roles in different developmental stages of spermatogenesis and testicular development. Five genes with ...
Development of the blood supply to the growth cartilage of the medial femoral condyle of foals.
Equine veterinary journal    April 1, 2020   Volume 53, Issue 1 134-142 doi: 10.1111/evj.13256
Wormstrand BH, Fjordbakk CT, Griffiths DJ, Lykkjen S, Olstad K.Growth cartilage is found in the articular-epiphyseal cartilage complex (AECC) and the physis. It has a temporary blood supply organised as end arteries. Vascular failure is associated with osteochondrosis, but infection can also obstruct vessels. The location of bacteria was recently compared to arterial perfusion, and the results indicated that they were located in the distal tips of AECC end arteries. Systematic perfusion studies were not available for comparison to the infected physes. Further studies may improve our understanding of infections and other pathologies. Objective: To describe...
Speed of in vitro embryo development affects the likelihood of foaling and the foal sex ratio.
Reproduction, fertility, and development    March 17, 2020   Volume 32, Issue 5 468-473 doi: 10.1071/RD19298
Claes A, Cuervo-Arango J, Colleoni S, Lazzari G, Galli C, Stout TA.The success of invitro embryo production (IVEP) in horses has increased considerably during recent years, but little is known about the effect of the speed of invitro embryo development. Blastocysts (n=390) were produced by intracytoplasmic sperm injection of IVM oocytes from warmblood mares, cryopreserved, thawed and transferred into recipient mares on Days 3, 4, 5 or 6 after ovulation. The time required for invitro-produced (IVP) embryos to reach the blastocyst stage was recorded (Day 7 vs Day 8). The likelihood of foaling was affected by the speed of invitro embryo development and recipient...
Immunological memory and tolerance at the maternal-fetal interface: Implications for reproductive management of mares.
Theriogenology    February 28, 2020   Volume 150 432-436 doi: 10.1016/j.theriogenology.2020.02.043
Antczak DF.The development of placentation that coincided with the evolution of mammals presented new challenges to the transmission of life from one generation to the next, particularly with regard to the possibility of maternal immunological recognition and destruction of the developing conceptus. The balance between immunity and tolerance dominates the immunological relationship between mother and fetus during mammalian pregnancy, and the focal point of this relationship lies at the interface between the trophoblast cells that comprise the outermost layer of the placenta and the maternal endometrial t...
Impact of the corpus luteum on survival of the developing embryo and early pregnancy in mares.
Theriogenology    February 19, 2020   Volume 150 374-381 doi: 10.1016/j.theriogenology.2020.02.011
Pinto CRF.It has been more than a hundred years that studies aiming to elucidate the processes involved in cyclicity and pregnancy pointed out the requirement of ovaries and corpora lutea for embryo survival and pregnancy establishment. For horses, luteal progesterone is essential for pregnancy only during the first trimester. This progestational support is complex among domestic animals as ovarian luteal function is further enhanced by the LH-action role of equine chorionic gonadotropin (eCG) starting ∼ on Day 35 of pregnancy. Increased eCG secretion leads to the formation of supplementary corpora ...
Evidence of five digits in embryonic horses and developmental stabilization of tetrapod digit number.
Proceedings. Biological sciences    February 5, 2020   Volume 287, Issue 1920 20192756 doi: 10.1098/rspb.2019.2756
Kavanagh KD, Bailey CS, Sears KE.Previous work comparing the developmental mechanisms involved in digit reduction in horses with other mammals reported that horses have only a 'single digit', with two flanking metapodials identified as remnants of digit II and IV. Here we show that early embryos go through a stage with five digit condensations, and that the flanking splint metapodials result from fusions of the two anterior digits I and II and the two posterior digits IV and V, in a striking parallel between ontogeny and phylogeny. Given that even this most extreme case of digit reduction exhibits primary pentadactyly, we re...
Maturation of the equine medial femoral condyle osteochondral unit.
Osteoarthritis and cartilage open    January 27, 2020   Volume 2, Issue 1 100029 doi: 10.1016/j.ocarto.2020.100029
Lemirre T, Santschi E, Girard C, Fogarty U, Chapuis L, Richard H, Beauchamp G, Laverty S.The juvenile equine medial femoral condyle (MFC) is frequently affected with radiographic changes (sclerosis and subchondral lucencies) that arise at a similar site to juvenile osteochondritis dissecans (JOCD) in children. There is little information on maturation of the MFC. To describe the normal development of the equine MFC osteochondral unit from birth to 2 years. Unassigned: Micro CT, histology and immunohistochemistry were performed on healthy equine MFCs (n = 29) at sites where lesions occur. Parameters assessed included: cartilage thickness; the epiphyseal growth plate cartilage org...
Embryo development after vitrification of immature and in vitro-matured equine oocytes.
Cryobiology    January 19, 2020   Volume 92 251-254 doi: 10.1016/j.cryobiol.2020.01.014
Angel D, Canesin HS, Brom-de-Luna JG, Morado S, Dalvit G, Gomez D, Posada N, Pascottini OB, Urrego R, Hinrichs K, Velez IC.Effects of meiotic stage and cumulus status on development of equine oocytes after vitrification was evaluated. Immature oocytes with corona radiata (IMM); in vitro-matured oocytes with corona radiata (MAT CR+); and in vitro-matured oocytes denuded of cumulus (MAT CR-) were vitrified using the Cryotech® method. Warming medium was equilibrated either in 5% CO2 or Air. IMM oocytes underwent in vitro maturation after warming. Recovery, survival, and maturation rates, and cleavage and blastocyst rates after ICSI, were evaluated. Recovery was higher for oocytes warmed in CO2- than Air-equilibrated...
Identification of Novel lncRNAs Differentially Expressed in Placentas of Chinese Ningqiang Pony and Yili Horse Breeds.
Animals : an open access journal from MDPI    January 11, 2020   Volume 10, Issue 1 119 doi: 10.3390/ani10010119
Pu Y, Zhang Y, Zhang T, Han J, Ma Y, Liu X.As a nutrient sensor, the placenta plays a key role in regulating fetus growth and development. Long non-coding RNAs (lncRNAs) have been shown to regulate growth-related traits. However, the biological function of lncRNAs in horse placentas remains unclear. To compare the expression patterns of lncRNAs in the placentas of the Chinese Ningqiang (NQ) and Yili (YL) breeds, we performed a transcriptome analysis using RNA sequencing (RNA-seq) technology. NQ is a pony breed with an average adult height at the withers of less than 106 cm, whereas that of YL is around 148 cm. Based on 813 million high...
Physiological development of the equine fetus during late gestation.
Equine veterinary journal    December 17, 2019   Volume 52, Issue 2 165-173 doi: 10.1111/evj.13206
Fowden AL, Giussani DA, Forhead AJ.In many species, the pattern of growth and physiological development in utero has an important role in determining not only neonatal viability but also adult phenotype and disease susceptibility. Changes in fetal development induced by a range of environmental factors including maternal nutrition, disease, placental insufficiency and social stresses have all been shown to induce adult cardiovascular and metabolic dysfunction that often lead to ill health in later life. Compared to other precocious animals, much less is known about the physiological development of the fetal horse or the longer-...
Assessing equine embryo developmental competency by time-lapse image analysis.
Reproduction, fertility, and development    November 25, 2019   Volume 31, Issue 12 1840-1850 doi: 10.1071/RD19254
Brooks KE, Daughtry BL, Metcalf E, Masterson K, Battaglia D, Gao L, Park B, Chavez SL.The timing of early mitotic events during preimplantation embryo development is important for subsequent embryogenesis in many mammalian species, including mouse and human, but, to date, no study has closely examined mitotic timing in equine embryos from oocytes obtained by ovum pick-up. Here, cumulus-oocyte complexes were collected by transvaginal follicular aspiration, matured invitro and fertilised via intracytoplasmic sperm injection. Each fertilised oocyte was cultured up to the blastocyst stage and monitored by time-lapse imaging for the measurement of cell cycle intervals and identifica...
Time of first polar body extrusion affects the developmental competence of equine oocytes after intracytoplasmic sperm injection.
Reproduction, fertility, and development    November 21, 2019   Volume 31, Issue 12 1805-1811 doi: 10.1071/RD19248
Rodríguez MB, Gambini A, Clérico G, Ynsaurralde-Rivolta AE, Briski O, Largel H, Sansinena M, Salamone DF.Assisted reproduction techniques (ARTs) have become widespread in the equine breeding industry. In particular, the combination of oocyte recovery from live mares followed by IVM and intracytoplasmic sperm injection (ICSI) has increased markedly among the ARTs used with valuable or low-fertility animals. There is currently no consensus among research groups regarding the optimal oocyte maturation period to produce high-quality embryos. In this study, we report the maturation dynamics of equine oocytes at different time points, from 20 to 40h (Experiment 1). In addition, in Experiment 2, equine ...
Characterization of the placental transcriptome through mid to late gestation in the mare.
PloS one    November 14, 2019   Volume 14, Issue 11 e0224497 doi: 10.1371/journal.pone.0224497
Loux SC, Dini P, El-Sheikh Ali H, Kalbfleisch T, Ball BA.The placenta is a dynamic organ which undergoes extensive remodeling throughout pregnancy to support, protect and nourish the developing fetus. Despite the importance of the placenta, very little is known about its gene expression beyond very early pregnancy and post-partum. Therefore, we utilized RNA-sequencing to characterize the transcriptome from the fetal (chorioallantois) and maternal (endometrium) components of the placenta from mares throughout gestation (4, 6, 10, 11 m). Within the endometrium, 47% of genes changed throughout pregnancy, while in the chorioallantois, 29% of genes under...
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