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.
Paris DB, Kuijk EW, Roelen BA, Stout TA.Real-time quantitative PCR (qPCR) is invaluable for investigating changes in gene expression during early development, since it can be performed on the limited quantities of mRNA contained in individual embryos. However, the reliability of this method depends on the use of validated stably expressed reference genes for accurate data normalisation. The aim of the present study was to identify and validate a set of reference genes suitable for studying gene expression during equine embryo development. The stable expression of four carefully selected reference genes and one developmentally regula...
Smits K, Goossens K, Van Soom A, Govaere J, Hoogewijs M, Peelman LJ.In vitro-produced (IVP) equine blastocysts can give rise to successful pregnancies, but their morphology and developmental rate differ from those of in vivo-derived equine blastocysts. The aim of the present study was to evaluate this difference at the genetic level. Suppression subtractive hybridisation (SSH) was used to construct a cDNA library enriched for transcripts preferentially expressed in in vivo-derived equine blastocysts compared with IVP blastocysts. Of the 62 different genes identified in this way, six genes involved in embryonic development (BEX2, FABP3, HSP90AA1, MOBKL3, MCM7 a...
Desmarais JA, Demers SP, Suzuki J, Laflamme S, Vincent P, Laverty S, Smith LC.Although putative horse embryonic stem (ES)-like cell lines have been obtained recently from in vivo-derived embryos, it is currently not known whether it is possible to obtain ES cell (ESC) lines from somatic cell nuclear transfer (SCNT) and parthenogenetic (PA) embryos. Our aim is to establish culture conditions for the derivation of autologous ESC lines for cell therapy studies in an equine model. Our results indicate that both the use of early-stage blastocysts with a clearly visible inner cell mass (ICM) and the use of pronase to dissect the ICM allow the derivation of a higher proportion...
Mlodawska W, Slomczynska M.Ovarian steroidogenesis from the neonatal to pubertal period in horses is poorly understood. This study was designed to immunolocalize cytochrome P450 aromatase in the ovarian follicles of slaughtered fillies ages approximately (I) 6-9 mo (5 mm in diameter. Staining intensity was dependent on the size and morphology of the follicle. In nonatretic follicles 5-10 mm in diameter, the reaction was weak and heterogeneous, while most intense staining was observed in preovulatory follicles. In follicles (diameter <20 mm) in the groups <10MF and 1YF, the reaction was less intense than in adult m...
De Lorenzi L, Genualdo V, Perucatti A, Pia Di Meo G, Molteni L, Iannuzzi L, Parma P.R-spondins constitute a recently discovered small family of growth factors, and the evidence of their role in several developmental pathways is growing fast. In this work we describe the chromosomal location of the four RSPO genes in the donkey. Using horse BACs, we localized RSPO1 on EAS 5q23, RSPO2 on EAS 12q13, RSPO3 on EAS 24q26, and RSPO4 on EAS 15p13. Moreover, RSPO2, RSPO3, and RSPO4 are the first genes mapped on donkey chromosomes 12, 24, and 15, respectively.
Veronesi MC, Villani M, Wilsher S, Contri A, Carluccio A.The aim of the study was to compare horse and donkey placentae using stereological techniques. Term placentae were collected at spontaneous foaling from seven Thoroughbred mares, seven pony mares, and six jenny donkeys. Maternal and foal weights were recorded and the mass, volume, and gross area of each allantochorion was also recorded. Ten random biopsies were recovered and processed for light microscopy from which the surface density of the microcotyledons (S(v)) and the total microscopic area of fetomaternal contact were calculated stereologically. Gestation length was longer in the donkeys...
Walter I, Tschulenk W, Budik S, Aurich C.The present study gives a detailed ultrastructural description of equine conceptuses at Day 14 (n = 2) and Day 16 (n = 3) after ovulation. Whereas on Day 14 only primitive structures were seen, on Day 16 neurulation and formation of mesodermal somites had taken place. The ectoderm of the embryo itself and the surrounding trophoblast ectodermal cells were characterised by specific cell surface differentiations. At the embryonic ectodermal cell surface (14 and 16 days) remarkable protruded and fused cytoplasmic projections were seen, typically associated with macropinocytotic events involved in ...
Wilsher S, Clutton-Brock A, Allen WR.A total of 78 day 10 horse embryos were transferred non-surgically to recipient mares that had ovulated 9, 7, 6, 5, 4, 3, 2 or 1 day after (negative asynchrony), on the same day (synchronous), or 2 or 4 days before (positive asynchrony) the donor (n=6 or 8 mares per group). Pregnancy rates between 100% (6/6) and 63% (5/8) were seen in recipient mares that were between +2 and -6 days asynchronous. Embryo survival to the heartbeat stage declined in recipients that were -7 days asynchronous and no embryos survived in recipients that were -9 days asynchronous. Irrespective of uterine asynchrony, c...
Allen WR, Wilsher S.Constant, self induced mobility throughout the uterine lumen between days 6 and 17 after ovulation, complete envelopment by a self-secreted glycoprotein capsule between days 7 and 30 and 'injection' of specialised, gonadotrophin-secreting trophoblast cells into the maternal endometrium at days 35-37 are three unusual aspects of equine embryogenesis. The outer trophoblast layer of the allantochorion finally establishes a stable, microvillous contact with the lumenal epithelium of the endometrium around days 40-42 and placentation commences thereafter. The allantochorion elongates steadily until...
Padua MB, Kowalski AA, Cañas MY, Hansen PJ.Uterine serpins (USs), designated as SERPINA14, are expressed in the endometrium in response to progesterone. All species identified as having USs exhibit epitheliochorial placentation and are in the Ruminantia and Suidae orders of the Laurasiatheria superorder. The objective was to identify US genes in species within and outside Laurasiatheria and evaluate whether evolution of the US gene was associated with development of the epitheliochorial placenta. Through queries of nucleotide and genomic databases, known US genes were identified (caprine, bovine, porcine, water buffalo), and new US cod...
Asher RJ, Bennett N, Lehmann T.An unprecedented level of confidence has recently crystallized around a new hypothesis of how living placental mammals share a pattern of common descent. The major groups are afrotheres (e.g., aardvarks, elephants), xenarthrans (e.g., anteaters, sloths), laurasiatheres (e.g., horses, shrews), and euarchontoglires (e.g., humans, rodents). Compared with previous hypotheses this tree is remarkably stable; however, some uncertainty persists about the location of the placental root, and (for example) the position of bats within laurasiatheres, of sea cows and aardvarks within afrotheres, and of der...
Choi YH, Harding HD, Hartman DL, Obermiller AD, Kurosaka S, McLaughlin KJ, Hinrichs K.The reported patterns of trophectodermal expression of POU5F1 protein in blastocysts vary among species, and are possibly related to the differences in placental growth and function. This study investigated the pattern of embryonic POU5F1 expression in the horse, a species with delayed placental formation. Immature equine oocytes expressed POU5F1 protein in the cytoplasm and nucleus. Staining for POU5F1 protein in in vitro-produced (IVP) embryos decreased to day 5 of culture, then the nuclear staining increased to day 7. IVP day-7 to -11 blastocysts showed POU5F1 staining in nuclei throughout ...
Tallmadge RL, McLaughlin K, Secor E, Ruano D, Matychak MB, Flaminio MJ.Many features of the equine immune system develop during fetal life, yet the naïve or immature immune state of the neonate renders the foal uniquely susceptible to particular pathogens. RT-PCR and immunohistochemical experiments investigated the progressive expression of developmental B cell markers and immunoglobulins in lymphoid tissues from equine fetus, pre-suckle neonate, foal, and adult horses. Serum IgM, IgG isotype, and IgA concentrations were also quantified in pre-suckle foals and adult horses. The expression of essential B cell genes suggests active development and gene recombinati...
Brianti E, Giannetto S, Traversa D, Chirgwin SR, Shakya K, Klei TR.A mixed population of equine cyathostomin (Nematoda, Strongyloidea) infective third stage larvae (L3) was cultured in vitro using a cell-free medium. Some L3 were cultured immediately after Baermann collection from fecal cultures, while others were kept in water at 4 degrees C for 7 days before initiating the in vitro cultures. Cultures were examined daily for viability. At days 2, 7, 14 and 21 larvae were collected for identification of developmental stage and morphological changes, using both light and scanning electron microscopy. Larvae were classified as early L3 (EL3), developing L3 (DL3...
Manso Filho HC, McKeever KH, Gordon ME, Manso HE, Lagakos WS, Wu G, Watford M.Glutamine is concentrated within skeletal muscle, where it has been proposed to play a regulatory role in maintaining protein homeostasis. The work presented here addressed the hypothesis that glutamine would be the most abundant free alpha-AA in plasma and skeletal muscle in the foal during the first year of life. Glycine, however, was the most abundant free alpha-AA in plasma at birth and between 3 and 12 mo of age. The concentration of glutamine, the second most abundant AA at birth, increased through the first 7 d (P < 0.05) and then returned to values similar to those at birth. This re...
Henry S, Richard-Yris MA, Tordjman S, Hausberger M.The neonatal period in humans and in most mammals is characterized by intense mother-young interactions favoring pair bonding and the adaptation of neonates to their new environment. However, in many post-delivery procedures, human babies commonly experience combined maternal separation and intense handling for about one hour post-birth. Currently, the effects of such disturbances on later attachment and on the development of newborns are still debated: clearly, further investigations are required. As animals present good models for controlled experimentation, we chose domestic horses to inves...
Lepeule J, Bareille N, Robert C, Ezanno P, Valette JP, Jacquet S, Blanchard G, Denoix JM, Seegers H.Developmental Orthopaedic Disease (DOD) in limb joints of horses is frequent and is a common cause of pain and lameness. DOD is a multifactorial disease involving genetics, growth, feeding practices and exercise conditions leading to joint injuries. However, there is no clear understanding of the contribution of each factor. The aim of this cohort study was to assess the adjusted effects of breed and gender, growth, feeding practices and exercise conditions on the prevalence of DOD in the limbs of foals at weaning and on the prevalence restricted to osteochondrosis (OC). Twenty-one farms in No...
Wilsher S, Allen WR.Eight day 10 horse embryos were transferred non-surgically to recipient mares that had ovulated 7 days after the donors. The embryonic vesicle was seen ultrasonographically in all eight recipients, and three out of eight (38%) of the vesicles developed an embryo proper with a beating heart. Conceptus expansion was initially slower than that in control mares but continued until day 22 (recipient day 15). Time of fixation of the vesicle was related to its diameter, rather than uterine stage. Although the embryo proper first appeared ultrasonographically on day 22, as normal, it grew more slowly ...
Lode A, Bernhardt A, Kroonen K, Springer M, Briest A, Gelinsky M.The application of bone graft substitutes with osteoinductive properties is of high importance for the repair of large bone defects. COLLOSS E, a protein lyophilizate extracted from equine long bones, exhibits an osteoinductive potential which has been proven in several studies. In this work, a mechanically stable, but biodegradable support for COLLOSS E has been developed aiming at a bone graft substitute that retains shape and size when coming in contact with body fluids. Mineralization of collagen type I, isolated from horse tendon, resulted in a stable collagen hydroxyapatite nanocomposite...
de Mestre AM, Miller D, Roberson MS, Liford J, Chizmar LC, McLaughlin KE, Antczak DF.The objective of this study was to identify transcription factors associated with differentiation of the chorionic girdle, the invasive form of equine trophoblast. The expression patterns of five transcription factors were determined on a panel of conceptus tissues from early horse pregnancy. Tissues from Days 15 through 46 were tested. Eomesodermin (EOMES), glial cells missing homologue 1 (GCM1), heart and neural crest derivatives expressed transcript 1 (HAND1), caudal type homeobox 2 (CDX2), and distal-less homeobox 3 (DLX3) were detected in horse trophoblast, but the expression patterns for...
Abd-Elnaeim MM, Derar IR, Wilsher S, Allen WR, Leiser R, Schuler G.The functions of placental oestrogens during equine pregnancy are still unclear. Yet, they may act predominantly as local regulators of growth and differentiation in the microplacentomes. Thus, expression patterns of oestrogen receptors (ERs) alpha and beta were investigated in the microcotyledonary placenta from pregnant mares at 110, 121, 179, 199 and 309 days of gestation by immunohistochemistry. In microplacentomes, both the ER isoforms were detected in trophoblast (T) cells, chorionic villous stroma (FS), microcaruncular epithelium (ME) and microcaruncular stroma (MS). Proportions of posi...
Carter AM.Recently a fossil of one of the earliest jawed fishes was found with a fetal skeleton and the remains of a cord. It was from the Devonian period and takes the history of vertebrate placentation back to 380 million years ago. This and later fossil evidence for viviparity in marine reptiles and early mammals is reviewed. Of particular interest are the fossils of horses as they document that a reproductive strategy with a single precocial newborn was evolved early on. In one instance there is sufficient representation of soft tissue to imply that early horses had a diffuse placenta, much as had b...
Manso Filho HC, Costa HE, Wu G, McKeever KH, Watford M.In most mammalian species the developing fetus utilizes large amounts of glutamine derived both from the maternal circulation and synthesized de novo in the placenta. The present study was designed to determine the role of the placenta in glutamine synthesis in the horse. The placentae from eight Standardbred mares were sampled immediately after parturition together with additional tissues obtained at necropsy from three Standbred mares during diestrous. Glutamine synthetase protein was detectable in the non-pregnant horn of the placenta in amounts similar to those seen in gluteus muscle, but ...
Galli C, Lagutina I, Duchi R, Colleoni S, Lazzari G.The cloning of equids was achieved in 2003, several years after the birth of Dolly the sheep and also after the cloning of numerous other laboratory and farm animal species. The delay was because of the limited development in the horse of more classical-assisted reproductive techniques required for successful cloning, such as oocyte maturation and in vitro embryo production. When these technologies were developed, the application of cloning also became possible and cloned horse offspring were obtained. This review summarizes the main technical procedures that are required for cloning equids an...
Olstad K, Ytrehus B, Ekman S, Carlson CS, Dolvik NI.The developmental pattern of the cartilage canal blood supply to epiphyseal growth cartilage has been linked to osteochondrosis (OC) in the tarsus of foals. This pattern has not yet been described in the distal femur, another site frequently affected by OC. Objective: To describe the developmental pattern of the blood supply to the distal femoral epiphyseal growth cartilage in 8 Standardbred foals age 0-7 weeks. Methods: One foal was sacrificed weekly from birth to age 7 weeks (n=8) to undergo a barium perfusion procedure to demonstrate vessels within cartilage canals of one hindlimb. The dist...
Lecocq M, Girard CA, Fogarty U, Beauchamp G, Richard H, Laverty S.The earliest osteochondrosis (OC) microscopic lesion reported in the literature was present in the femorotibial joint of a 2-day-old foal suggesting that OC lesions and factors initiating them may arise prior to birth. Objective: To examine the developing equine epiphysis to detect histological changes that could be precursors to OC lesions. Methods: Osteochondral samples from 21 equine fetuses and 13 foals were harvested from selected sites in the scapulohumeral, humeroradial, metacarpophalangeal, femoropatellar, femorotibial, tarsocrural and metatarsophalangeal joints. Sections were stained ...
Carluccio A, Panzani S, Tosi U, Riccaboni P, Contri A, Veronesi MC.This study was designed to establish the morphological features of the placenta of the Martina Franca jenny. Ten placentas were harvested at the time of foal delivery and examined both for gross and histological characteristics. The following factors were determined: the total weight and volume of the placenta and its components, the surface area of the allantochorion, umbilical cord length and site of insertion, and the diameter of the umbilical cord vessels and vascular pattern type. The weight of the placenta was similar to previously reported for ponies, and represented 12% of foal birth w...
Gerstenberg C, Allen WR, Stewart F.Placentation involves considerable growth and reorganization of both maternal and fetal tissues. In this investigation, immunohistochemical localization of the proliferation marker Ki-67 antigen was used to monitor cell division during placentation in mares. Endometrial biopsies were obtained from eight mares between day 14 and day 26 of pregnancy and from eight anoestrous mares that had been treated with various combinations of progesterone and oestrogen. Samples of endometrium and fetal membranes were obtained from 19 mares carrying normal horse conceptuses between day 30 and day 250 of gest...
Choi I, Collante WR, Simmen RC, Simmen FA.Pig blastocysts exhibit a transient period of estrogen production at periimplantation, with a second, more sustained period of estrogen synthesis occurring in endometrium and placenta at later pregnancy. Previously we reported the isolation of cDNA clones encoding a novel isoform of cytochrome P450 aromatase (the terminal enzyme in the estrogen biosynthetic pathway) from porcine periimplantation blastocysts. The present study investigated pregnancy-associated expression, in blastocysts and maternal reproductive tract tissues of this and an additional mRNA transcript encoding a distinct P450 ar...
Rigoglio NN, Barreto RS, Favaron PO, Jacob JC, Smith LC, Gastal MO, Gastal EL, Miglino MA.The neural system is one of the earliest systems to develop and the last to be fully developed after birth. This study presents a detailed description of organogenesis of the central nervous system (CNS) at equine embryonic/fetal development between 19 and 115 days of pregnancy. The expression of two important biomarkers in the main structure of the nervous system responsible for neurogenesis in the adult individual, and in the choroid plexus, was demonstrated by Nestin and glial fibrillary acid protein (GFAP) co-labeling. In the 29th day of pregnancy in the undifferentiated lateral ventricle...
Price JS, Jackson BF, Gray JA, Harris PA, Wright IM, Pfeiffer DU, Robins SP, Eastell R, Ricketts SW.This study describes longitudinal changes in serum levels of biochemical markers of bone cell activity in a group of 24 thoroughbred foals from birth to 18 months of age. The markers of bone formation included the type I collagen carboxy-terminal propeptide (PICP), the bone-specific isoenzyme of alkaline phosphatase (BAP), and osteocalcin (OC). Levels of the cross-linked telopeptide of type I collagen (ICTP), a marker of bone resorption, and the N-terminal propeptide of type III collagen (PNIIIP), a marker of soft tissue turnover, were also measured. Levels of all markers fell significantly be...
Liu Y, Du M, Li X, Zhang L, Zhao B, Wang N, Dugarjaviin M.This study aimed to investigate differences in testicular tissue morphology, gene expression, and marker genes between sexually immature (1-year-old) and sexually mature (10-year-old) Mongolian horses. The purposes of our research were to provide insights into the reproductive physiology of male Mongolian horses and to identify potential markers for sexual maturity. The methods we applied included the transcriptomic profiling of testicular cells using single-cell sequencing techniques. Our results revealed significant differences in tissue morphology and gene expression patterns between the tw...
Mady EA, Osuga H, Toyama H, El-Husseiny HM, Inoue R, Murase H, Yamamoto Y, Nagaoka K.The gut microbiota (GM) is essential for mammalian health. Although the association between infant GM and breast milk (BM) composition has been well established in humans, such a relationship has not been investigated in horses. Hence, this study was conducted to analyze the GM formation of foals during lactation and determine the presence of low-molecular-weight metabolites in mares' BM and their role in shaping foals' GM. The fecal and BM samples from six pairs of foals and mares were subjected to 16S ribosomal RNA metagenomic and metabolomic analyses, respectively. The composition of foal G...
Choi YH, Love CC, Varner DD, Hinrichs K.This study was conducted to evaluate the effects of thawing, division into aliquots and refreezing on fertilizing capacity (ability to support embryo development after intracytoplasmic sperm injection; ICSI) of frozen stallion semen. Frozen semen from a fertile stallion was thawed, diluted 1:100 with freezing extender, and refrozen (2F treatment). Control semen was frozen only once. In vitro matured equine oocytes were injected with: (1) motile control spermatozoa; (2) motile 2F spermatozoa; (3) non-motile 2F spermatozoa; or (4) non-motile 2F spermatozoa, followed by injection of sperm extract...
Vegas AR, Podico G, Canisso IF, Bollwein H, Fröhlich T, Bauersachs S, Almiñana C.During initial maternal recognition of pregnancy (MRP), the equine embryo displays a series of unique events characterized by rapid blastocyst expansion, secretion of a diverse array of molecules, and transuterine migration to interact with the uterine surface. Up to date, the intricate transcriptome and proteome changes of the embryo underlying these events have not been critically studied in horses. Thus, the objective of this study was to perform an integrative transcriptomic (including mRNA, miRNAs, and other small non-coding RNAs) and proteomic analysis of embryos collected from days 10 t...
González-Angulo A, Hernández-Jáuregui P, Martínez-Zedilo G.Light and electron microscopic studies of the gonads of the fetal horse have shown that, in their hypertrophic condition which begins during the 3rd month, the interstitial cells contain large amounts of smooth endoplasmic reticulum, suggesting a secretory activity. Hydroxylating activity which was cytochrome P-450-dependent was observed in the fetal testis and may be involved in steroid biosynthesis.
Green J, Xie S, Gan X, Roberts RM.This manuscript describes the cloning of a novel aspartic proteinase expressed in the placenta of the horse (order Perrisodactyla). Evidence for similar genes in the cat (Carnivora) and ruminants (Artiodactyla), indicates that these molecules have been conserved within widely divergent species with distinct types of placentation. Since ePAG is produced by the outer cell layer (trophoblast) of the placenta, it can tentatively be grouped with the pregnancy-associated glycoproteins (PAG) of cattle, sheep, and pig. The high sequence identity that ePAG shares with pepsinogens as well as the PAG, in...
Reinholt BM, Bradley JS, Jacobs RD, Ealy AD, Johnson SE.Rapid morphological and gene expression changes occur during the early formation of a mammalian blastocyst. Critical to successful retention of the blastocyst and pregnancy is a functional trophectoderm (TE) that supplies the developing embryo with paracrine factors and hormones. The contribution of TE conformational changes to gene expression was examined in equine induced trophoblast (iTr) cells. Equine iTr cells were cultured as monolayers or in suspension to form spheres. The spheres are hollow and structurally reminiscent of native equine blastocysts. Total RNA was isolated from iTr monol...
Smyth GB.Lengths of small intestine, caecum, ascending colon and descending colon were measured in intestinal tracts from 103 foetuses, foals and adult horses. Intestinal tracts from 21 of the horses were measured at varying intervals after death. Total intestinal length increased markedly from mid-gestation to one year of age, then showed little increment up to 35 years. Small intestinal length increased most rapidly in the first month of life. Caecal length increased most rapidly from one to six months and length of ascending colon increased most rapidly between one week and one year. Descending colo...
Jones CJP, Allen WRT, Wilsher S.In this study, we compare glycosylation at the fetomaternal interface in 3 equine species: horse, donkey and zebra, all of which can interbreed to produce hybrids, to assess their glycan similarities and differences. Methods: Sections cut from 3 specimens of horse (Equus caballus) placenta (50, 200 and 280 days gestation), one donkey (Equus asinus) placenta (65 cm crown-rump length) and 5 specimens of zebra (Equus quagga) placentae (81-239 days gestation) were stained with a panel of 24 biotinylated lectins using an avidin-peroxidase revealing system. Results: There were only slight quantitat...
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...
Ousey JC, Kölling M, Newton R, Wright M, Allen WR.Aged mares with endometrosis suffer higher rates of pregnancy loss than young mares, due to poor placental development. Reduced uterine blood supply may be one contributory factor. Objective: To measure uterine artery (UA) blood flow and other Doppler indices throughout pregnancy and compare placental and foal development in young mares and aged mares. Methods: Thoroughbred mares were grouped according to age and endometrial biopsy score: 1) 6 young mares (mean age 7.3 years, Category ); 2) 6 aged mares (mean age 18.3 years, Category II). Vascular pathology was nil or mild except in one aged m...
Prieto JMB, Tallmadge RL, Felippe MJB.Identification and classification of B cell subpopulations has been shown to be challenging and inconsistent among different species. Our study tested aspects of ontogeny, phenotype, tissue distribution, and function of equine CD5 B cells, which represented a greater proportion of B cells early in development and in the peritoneal cavity. CD5 and CD5 B cells differentially expressed B cell markers (CD2, CD21, IgM) measured using flow cytometry, but similar mRNA expression of signature genes (DGKA, FGL2, PAX5, IGHM, IL10) measured using quantitative RT-PCR. Sequencing lambda light chain segment...
Colchen S, Battut I, Fiéni F, Tainturier D, Siliart B, Bruyas JF.Equine embryos were collected at exactly 156 +/- 0.5 (n=8) and 168 +/- 0.5 h (n=11) after ovulation. The embryos were fixed in glutaraldehyde, sectioned serially and observed using light microscopy. In the 156 h group, all embryos were early blastocysts except for one, which was a morula. The morula and one early blastocyst had no capsule. The capsules of the other embryos were thin. The mean +/- SD total number of cells was 275 +/- 105 (range 117-417). The mean +/- SD proportions of mitotic and pycnotic cells were 2.5 +/- 1.2 and 1.1 +/- 1.8%, respectively, and there were no differences betwe...
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-...
Hinrichs K, DiGiorgio LM.A technique was developed in which immature horse oocytes, obtained from slaughterhouse specimens, were transferred to the pre-ovulatory follicle of a mare in vivo, with resulting oocyte maturation, ovulation, fertilization and embryo development. Oocytes were collected from all follicles greater than 3 mm, and were classified as immature, maturing, expanded or denuded. The transfers were performed in the standing, tranquilized mare. The ovary containing the pre-ovulatory follicle was grasped per rectum. A trochar and cannula were placed through the abdominal wall in the flank area, ipsilatera...
Lennard SN, Stewart F, Allen WR.Placentation in equids involves two types of trophoblast: a minor invasive component, the chorionic girdle, that gives rise to transient endocrine structures known as endometrial cups, and a major non-invasive component, the allantochorion, that forms the diffuse, microcotyledonary placenta. Growth factors are likely to be important in controlling these complex events at implantation and this study describes the use of in situ hybridization and northern blotting techniques to monitor expression of insulin-like growth factor II (IGF-II) in the fetus and placenta of the horse (Equus caballus), u...
Manso Filho HC, Costa HE, Wu G, McKeever KH, Watford M.In most mammalian species the developing fetus utilizes large amounts of glutamine derived both from the maternal circulation and synthesized de novo in the placenta. The present study was designed to determine the role of the placenta in glutamine synthesis in the horse. The placentae from eight Standardbred mares were sampled immediately after parturition together with additional tissues obtained at necropsy from three Standbred mares during diestrous. Glutamine synthetase protein was detectable in the non-pregnant horn of the placenta in amounts similar to those seen in gluteus muscle, but ...
Padua MB, Kowalski AA, Cañas MY, Hansen PJ.Uterine serpins (USs), designated as SERPINA14, are expressed in the endometrium in response to progesterone. All species identified as having USs exhibit epitheliochorial placentation and are in the Ruminantia and Suidae orders of the Laurasiatheria superorder. The objective was to identify US genes in species within and outside Laurasiatheria and evaluate whether evolution of the US gene was associated with development of the epitheliochorial placenta. Through queries of nucleotide and genomic databases, known US genes were identified (caprine, bovine, porcine, water buffalo), and new US cod...
Scarlet D, Handschuh S, Reichart U, Podico G, Ellerbrock RE, Demyda-Peyrás S, Canisso IF, Walter I, Aurich C.It was the aim of this study to characterize the development of the gonads and genital ducts in the equine fetus around the time of sexual differentiation. This included the identification and localization of the primordial germ cell population. Equine fetuses between 45 and 60 days of gestation were evaluated using a combination of micro-computed tomography scanning, immunohistochemistry, and multiplex immunofluorescence. Fetal gonads increased in size 23-fold from 45 to 60 days of gestation, and an even greater increase was observed in the metanephros volume. Signs of mesonephros atrophy wer...
Choi YH, Velez IC, Macías-García B, Hinrichs K.In nuclear transfer (NT), exposure of donor cell chromatin to the ooplast cytoplasm may aid reprogramming; however, the length of exposure feasible is limited by the developmental life span of the oocyte. We examined the effect of duration of nucleus-cytoplasmic exposure before activation and of in vitro maturation (IVM) in equine NT. In experiment 1, 24 h IVM and a delay of 2, 5, or 8 h between reconstruction and activation yielded 4%, 15%, and 11% blastocysts, respectively. In experiment 2, a 5-h activation delay yielded 17% and 22% blastocysts with two donor cell lines. In experiment 3, usi...
Chavatte-Palmer P, Derisoud E, Robles M.Horses have been domesticated by man and historical information mostly associates horses with men. Nowadays, however, horse riding is essentially by women. Women are also very much involved in equine sciences, with a large contribution to the understanding of fetoplacental development. While highlighting the work of female scientists, this review describes the recent advances in equine fetoplacental studies, focusing on data obtained by new generation sequencing and progress on the understanding of the role of placental progesterone metabolites throughout gestation. A second emphasis is made o...
Kemper AM, Drnevich J, McCue ME, McCoy AM.Skeletogenesis is complex and incompletely understood. Derangement of this process likely underlies developmental skeletal pathologies. Examination of tissue-specific gene expression may help elucidate novel skeletal developmental pathways that could contribute to disease risk. Our aim was to identify and functionally annotate differentially expressed genes in equine neonatal and adult articular cartilage (AC) and subchondral bone (SCB). RNA was sequenced from healthy AC and SCB from the fetlock, hock, and stifle joints of 6 foals (≤4 weeks of age) and six adults (8-12 years of age). There w...
