Browsing by Subject "Embryo"
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- PublicationOpen Access3D model of the maternal-fetal interface: challenges, recent advances and beyond(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2023) Passaponti, Sofia; Luongo, Francesca Paola; Ietta, Francesca; Luddi, Alice; Piombon, PaolaEmbryo implantation is a complex and highly coordinated process that involves an intricate network of factors establishing intimate contact at the maternal-fetal interface. Knowledge of the human implantation process is compromised by both ethical issues, which do not allow the study of this process in vivo, and by the accuracy and reproducibility of in vitro models of human endometrium. Effective and reliable embryo implantation models are, therefore, necessary to mimic the molecular event cascade that occurs in vivo. 3D models are considered a new step to foster precision medicine and an advanced tool for the study of endometrial biology, endometrium associated diseases and to understand the complex mechanisms surrounding endometrium-embryo crosstalk. In this review we explore the various methods by which 3D cultures of endometrium and trophoblast can be created, exploring targets and applications of these in vitro models.
- PublicationOpen AccessAn overview on the diversity of cellular organelles during the germ cell cycle(Murcia : F. Hernández, 2010) Chuva de Sousa Lopes, Susana M.; Roelen, Bernard A.J.In mammals, germ cells undergo a longjourney from specification until sexual maturation.During this journey, which takes place during the entirelife cycle of mammals, the germ cells dynamicallychange their morphology, their expression profile andalso the number and character of their cellular bodies.The focus of this review will be the diversity of cellularorganelles present in the nucleus and cytoplasm at thedifferent phases of germ cell development. We discusshow these organelles associate and behave to form amultitude of bodies that have long been observed byscientists, and how their presence or absence is used tocharacterize different stages of germ cell development.These organelles include the female Barr body, polarbodies and Balbiani body; and the male sex body and chromatoid body. It is concluded thatcompartmentalization of organelles and molecules in thecytoplasm (in particular of mitochondria and RNAs) andof the sex chromosomes in the nucleus seems to beimportant for regulating germ cell developmentthroughout the life cycle
- PublicationOpen AccessComparative View on the Oviductal Environment during the Periconception Period.(2020) González-Brusi, Leopoldo; Algarra, Blanca; Izquierdo Rico, María José; Avilés Sánchez, Manuel; Jíménez Movilla, María; Moros Nicolás, Carla; Biología Celular e HistologíaThe oviduct plays important roles in reproductive events: sperm reservoir formation, final gamete maturation, fertilization and early embryo development. It is well known that the oviductal environment affects gametes and embryos and, ultimately, the health of offspring, so that in vivo embryos are better in terms of morphology, cryotolerance, pregnancy rates or epigenetic profile than those obtained in vitro. The deciphering of embryo–maternal interaction in the oviduct may provide a better understanding of the embryo needs during the periconception period to improve reproductive efficiency. Here, we perform a comparative analysis among species of oviductal gene expression related to embryonic development during its journey through the oviduct, as described to date. Cross-talk communication between the oviduct environment and embryo will be studied by analyses of the secreted or exosomal proteins of the oviduct and the presence of receptors in the membrane of the embryo blastomeres. Finally, we review the data that are available to date on the expression and characterization of the most abundant protein in the oviduct, oviductin (OVGP1), highlighting its fundamental role in fertilization and embryonic development.
- PublicationOpen AccessFish tales: The use of zebrafish xenograft human cancer cell models(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Drabsch, Yvette; Snaar Jagalska, Ewa; Ten Dijke, PeterAdvances in scientific techniques have provided researchers with exceptional new opportunities to identify and monitor changes between different cancer types, during different stages of progression, between individual tumor cells and in the surrounding stroma. The wealth of information that can be obtained from new scientific techniques places additional requirements on the conventional cancer models. New models that could be used to rapidly access the (potential) functional importance of newly identified (epi)genetic and proteomic changes and test the efficacy on emerging (combinatorial) therapies are desperately required. The distinctive characteristics of zebrafish are progressively being applied to create more relevant models of human diseases. Zebrafish embryos provide a powerful tool to develop functional cancer models. This is a tool that can be used from drug discovery and development to assessment of drug toxicity. This review will summarise the use of zebrafish xenograft models to study human cancers, and discuss the benefits and limitations of these models.
- PublicationOpen AccessIntrauterine Infusion of TGF-β1 Prior to Insemination, Alike Seminal Plasma, Influences Endometrial Cytokine Responses but Does Not Impact the Timing of the Progression of Pre-Implantation Pig Embryo Development(MDPI, 2021-02-17) Martínez, Cristina A.; Cambra, Josep M.; Lucas, Xiomara; Ferreira-Dias, Graça; Rodríguez-Martínez, Heriberto; Gil, María A.; Martínez, Emilio A.; Cuello, Cristina; Parrilla, Inmaculada; Medicina y Cirugía AnimalSeminal plasma (SP) in the female genital tract induces changes that affect multiple reproductive processes. One of the active components in SP is the transforming growth factor β1 (TGF-β1), which has major roles in embryo development and pregnancy. Embryo transfer (ET) technology is welcomed by the pig industry provided that embryo quality at embryo collection as well as the fertility and prolificacy of the recipients after the ET is increased. This study evaluated different intrauterine infusion treatments at estrus (40 mL of SP, TGF-β1 cytokine in the extender, or the extender alone (control)) by mimicking an ET scenario in so-called "donor" (inseminated) and "recipient" (uninseminated) sows. On day 6 (day 0-onset of estrus), all "donors" were laparotomized to determine their pregnancy status (presence and developmental stage of the embryos). In addition, endometrial explants were collected from pregnant "donors" and cyclic "recipients," incubated for 24 h, and analyzed for cytokine production. SP infusions (unlike TGF-β1 infusions) positively influenced the developmental stage of day 6 embryos. Infusion treatments differentially influenced the endometrial cytokine production, mainly in donors. We concluded that SP infusions prior to AI not only impacted the porcine preimplantation embryo development but also influenced the endometrial cytokine production six days after treatment, both in donors and recipients.
- PublicationOpen AccessPrimordial germ cell specification, the importance of being blimped(Murcia : F. Hernández, 2008) Chuva de Sousa Lopes, Susana M.; Roelen, Bernard A.J.In mouse embryos, the expression of Blimp1 has recently revealed a population of allocated primordial germ cell precursors 24 hours earlier than previously thought. Those ‘blimped’ precursors have been shown to give rise, by mitotic division, to germ cells only and no other cell lineages. Here, we try to understand the events that lead to Blimp1 expression in the primordial germ cell precursors and speculate on what can be the role of Blimp1 during primordial germ cell specification and gastrulation in the mouse. Finally, we discuss the possible involvement of Blimp1 in the two know modes of germ line segregation (epigenesis and preformation).
- PublicationOpen AccessProduction and characterization of a new monoclonal antibody, GR-ICOR-2, recognizing sarcomeric actin: analysis of the expression in the developing chick heart(Murcia : F. Hernández, 1994) Fernández, J.E.; Melguizo, C.; Prados, J.; Marchal, J.A.; Alvarez, L.; Aránega, A.We produced and characterized a specific monoclonal antibody (mAB) designated GR-ICOR-2. This mAb recognizes sarcomeric actin molecules (43 kDa) and was used in an immunohistochemical analysis of staining pattems in Harnburger and Hamilton's stages 18, 22 and 25 (HH 18, 22 and 25) embryonic chick hearts. Staining showed a mainly cytoplasmic distrubition in three regions: the atrioventricular (AV) canal cushion tissue, the primitive ventricle, and conal crests. In addition, this mAb-cross-reacted with rabbit and human cardiac and skeletal muscle tissue; but not with smooth muscle tissue.
- PublicationOpen AccessRevisión actual del contexto legal y ético de la experimentación con embriones humanos.(Universidad de Murcia, Servicio de Publicaciones, 2021) Guerrero Franco, Juan José; Hernández López, Antonio; Guardiola Cutillas, Alicia; Guzmán Almansa, MiriamEn este estudio se pretende hacer una revisión de la situación actual de la experimentación con embriones humanos. El derecho comparativo entre distintos países revela tres posturas legislativas principales: permisiva, restrictiva y prohibitiva. En cuanto a las influencias, se comenta el estatuto del embrión desde el punto de visto religioso y sociopolítico. Además, se abordan problemas éticos y dilemas actuales asociados, comenzando con el famoso Caso Brüstle y culminando con los últimos estudios de células quiméricas. La disparidad de posiciones legales y sociales, junto con los nuevos avances tecnológicos, deja en evidencia la necesidad de un consenso internacional que marque los límites de los ensayos con embriones y aclare la identidad del estatuto del embrión.
- PublicationOpen AccessSelectividad de las isoformas de 140-180 KDA de NCAM en la papila óptica del embrión de pollo(Murcia: Universidad de Murcia, Servicio de Publicaciones, 1997) Prada Oliveira, J. A.; Verástegui Escolano, G.; González Moreno, M; Pérez Ríos, N.; Fernández Trujillo, F. J.; Facultad de BiologíaDurante la embriogénesis de la retina del Gallus domesticus, los neuroblastos muestran variaciones temporales y espaciales de los residuos glucosilados a nivel de sus membranas plasmáticas. Se considera que estos componentes glucídicos están implicados en la organización de estructuras neurales inmaduras, mediante su participación en mecanismos de reconocimiento y adhesión celulares. Los componentes glucosilados van a formar parte de glucoproteínas convencionales, así como de un grupo de moléculas a las que genéricamente se han denominado moléculas de adhesión celular. La primera de estas moléculas aislada fue la neural cell adhesion molecule (NCAM), glucoproteína compuesta por un armazón proteico con diversas isoformas y una cadena más o menos variable de polímeros de ácidos siálicos. La combinación de técnicas convencionales histoquímicas, de lectinas e inmunocitoquímicas han permitido determinar que las moléculas de NCAM de 120 kDa presentan cadenas pobremente sializadas en estadíos intermedios del desarrollo, mientras en estadíos tempranos y finales los ácidos siálicos están más abundantemente representados en las isoformas de 140-180 kDa. Algunos autores han determinado que la isoforma de NCAM presente durante el desarrollo de la retina del Gallus domesticus es similar a la humana de 120 kDa. Nuestros resultados indican que las isoformas de 140-180 kDa sólo se presentan en las fibras del nervio óptico una vez que estas abandonan la papila óptica.
- PublicationOpen AccessSeminal Plasma Modifies the Transcriptional Pattern of the Endometrium and Advances Embryo Development in Pigs(2019-12-18) Martínez, Cristina A.; Cambra, Josep M.; Parrilla, Inmaculada; Roca, Jordi; Ferreira-Dias, Graça; Pallarés, Francisco J.; Lucas, Xiomara; Vázquez, Juan M.; Martínez, Emilio A.; Gil, María A.; Rodríguez-Martínez, Heriberto; Cuello, Cristina; Álvarez-Rodríguez, Manuel; Medicina y Cirugía AnimalBackground: Seminal plasma (SP) promotes sperm survival and fertilizing capacity, and potentially affects embryo development, presumably via specific signaling pathways to the internal female genital tract. Objectives: This study evaluated how heterologous SP, infused immediately before postcervical artificial insemination (AI) affected embryo development and the transcriptional pattern of the pig endometria containing embryos. Materials and Methods: Postweaning estrus sows (n = 34) received 40-mL intrauterine infusions of either heterologous pooled SP or Beltsville Thawing Solution (BTS; control) 30 min before AI of semen extended to 10% of homologous SP. Embryos (all sows) and endometrium samples (3 sows/group) were removed during laparotomy 6 days after the infusion of SP or BTS to morphologically evaluate the embryos to determine their developmental stage and to analyze the endometrial transcriptome using microarrays (PORGENE 1.0 ST GeneChip array, Affymetrix) followed by qPCR for further validation. Results: Embryo viability was equal between the groups (~93%), but embryo development was significantly (P < 0.05) more advanced in the SP-treated group compared to control. A total of 1,604 endometrium transcripts were differentially expressed in the SP group compared to the control group. An enrichment analysis showed an overrepresentation of genes and pathways associated with the immune response, cytokine signaling, cell cycle, cell adhesion, and hormone response, among others. Conclusions: SP infusions prior to AI positively impacted the preimplantation embryo development and altered the expression of the endometrial genes and pathways potentially involved in embryo development.
- PublicationOpen AccessSupplementation of bovine follicular fluid during in vitro maturation increases oocyte cumulus expansion, blastocyst developmental kinetics and blastocyst cell number.(2019-01-14) Lopes, Jordana S; Canha-Gouveia, Analuce; París-Oller, Evelynne; Coy Fuster, Pilar; Coy Fuster, Pilar; FisiologíaBovine follicular fluid (bFF) is the natural milieu for oocyte growth and development. However, its value as supplementation to in vitro maturation medium is still questioned due to inconsistent results. In this study we hypothesized that adding 10% of follicular fluid as well as heat treating it to inhibit the complement system, would produce higher quality embryos. To do so, experiments were conducted to compare the effect of bFF and heat-treated bFF (bFFin) on oocyte competence assessed by different parameters such as nuclear and cytoplasmic maturation, IVF efficiency, in vitro embryo development and embryo survivability post-vitrification. No differences on nuclear maturation nor cortical granules migration were observed but differences were found on oocyte’s cumulus cell expansion, with bFF group having the highest increase (79.0±3.7%). bFFin had a negative impact on IVF efficiency (58.6±3.2%), but no differences were found between bFF (62.9±3.2%) and control (72.8±3.0%). Although the cleavage and blastocyst rate were similar between groups, the day 6 embryo development rate was higher in bFFin group, suggesting an accelerated developmental kinetics. Hatched blastocysts from the bFF group showed a higher cell count than the control group (241.3±20.1 and 185.8± 10.0, respectively), and bFFin embryos showed values in between (214.9±14.0). No difference on survivability post-vitrification was found between groups, although the blastocyst stage had a significant impact on the survival rate across all groups. In conclusion, using bFF as supplementation to maturation medium showed a higher benefit when comparing to the standard supplementation by having oocytes with higher cumulus expansion rate, faster development of embryos and higher number of cells per embryo. Inactivation of bFF lowered IVF efficiency but didn’t compromise blastocyst development and quality.