scholarly journals Wwc2 is a novel cell division regulator during preimplantation mouse embryo lineage formation and oogenesis

Author(s):  
Giorgio Virnicchi ◽  
Pablo Bora ◽  
Lenka Gahurová ◽  
Andrej Šušor ◽  
Alexander W. Bruce

ABSTRACTFormation of a mature and hatching mouse blastocyst marks the end of the preimplantation development, whereby regulated cell cleavages culminate in the formation of three distinct lineages. We report dysregulated expression of Wwc2, an ill-characterised paralog of the Hippo-signalling activator Kibra/Wwc1, is specifically associated with cell autonomous deficits in embryo cell number and cell division abnormalities, typified by imbalanced daughter cell chromatin segregation. Division phenotypes are also observed during mouse oocyte meiotic maturation, as Wwc2 dysregulation blocks progression to the fertilisation competent stage of meiosis II metaphase arrest, characterised by spindle defects and failed Aurora-A kinase (AURKA) activation. Such cell division defects, each occurring in the absence of centrosomes, are fully reversible by expression of recombinant HA-epitope tagged WWC2, restoring activated oocyte AURKA levels. Additionally, clonal dysregulation implicates Wwc2 in maintaining the pluripotent late blastocyst stage epiblast lineage. Thus, Wwc2 is a novel regulator of meiotic and early mitotic cell divisions, and mouse blastocyst cell-fate.

Author(s):  
William C Lester ◽  
Taylor Johnson ◽  
Ben Hale ◽  
Nicholas Serra ◽  
Brian Elgart ◽  
...  

Abstract Aurora A kinase (AURKA) is an important regulator of cell division and is required for assembly of the mitotic spindle. We recently reported the unusual finding that this mitotic kinase is also found on the sperm flagellum. To determine its requirement in spermatogenesis, we generated conditional knockout animals with deletion of the Aurka gene in either spermatogonia or spermatocytes to assess its role in mitotic and postmitotic cells, respectively. Deletion of Aurka in spermatogonia resulted in disappearance of all developing germ cells in the testis, as expected given its vital role in mitotic cell division. Deletion of Aurka in spermatocytes reduced testis size, sperm count, and fertility, indicating disruption of meiosis or an effect on spermiogenesis in developing mice. Interestingly, deletion of Aurka in spermatocytes increased apoptosis in spermatocytes along with an increase in the percentage of sperm with abnormal morphology. Despite the increase in abnormal sperm, sperm from spermatocyte Aurka knockout mice displayed increased progressive motility. In addition, sperm lysate prepared from Aurka knockout animals had decreased protein phosphatase 1 (PP1) activity. Together, our results show that AURKA plays multiple roles in spermatogenesis, from mitotic divisions of spermatogonia to sperm morphology and motility.


1982 ◽  
Vol 35 (2) ◽  
pp. 187 ◽  
Author(s):  
GM Harlow ◽  
P Quinn

The culture conditions for the development in vitro of (C57BL/6 X CBA) F2 hybrid two-cell embryos to the blastocyst stage have been optimized. Commercially available pre-sterile disposable plastic culture dishes supported more reliable development than re-usable washed glass tubes. The presence of an oil layer reduced the variability in development. An average of 85 % of blastocysts developed from hybrid two-cell embryos cultured in drops of Whitten's medium under oil in plastic culture dishes in an atmosphere of 5% O2 : 5% CO2 : 90% N2 ? The time taken for the total cell number to double in embryos developing in vivo was 10 h, and in cultured embryos 17 h. Embryos cultured in vitro from the two-cell stage to blastocyst stage were retarded by 18-24 h in comparison with those remaining in vivo. Day-4 blastocysts in vivo contained 25-70 cells (mean 50) with 7-28 (mean 16) of these in the inner cell mass. Cultured blastocysts contained 19-73 cells (mean 44) with 8-34 (mean 19) of these in the inner cell mass. In the uterine environment, inner-cell-mass blastomeres divided at a faster rate than trophectoderm blastomeres and it is suggested that a long cell cycle is associated with terminal differentiation. Although cultured blastocysts and inner cell masses contained the same number of cells as blastocysts and inner cell masses in vivo, the rate of cell division in cultured inner cell masses was markedly reduced.


Zygote ◽  
2004 ◽  
Vol 12 (3) ◽  
pp. 205-213 ◽  
Author(s):  
Gabriela Il'ková ◽  
Pavol Rehák ◽  
Jarmila Veselá ◽  
štefan Čikoš ◽  
Dušan Fabian ◽  
...  

Serotonin is a neurotransmitter functioning also as a hormone and growth factor. To further investigate the biological role of serotonin during embryo development, we analysed serotonin localization as well as the expression of specific serotonin 5-HT1D receptor mRNA in mouse oocytes and preimplantation embryos. The functional significance of serotonin during the preimplantation period was examined by studying the effects of serotonin on mouse embryo development. Embryo exposure to serotonin (1 μM) highly significantly reduced the mean cell number, whereas lower concentrations of serotonin (0.1 μM and 0.01 μM) had no significant effects on embryo cell numbers. In all serotonin-treated groups a significant increase in the number of embryos with apoptotic and secondary necrotic nuclei was observed. Expression of serotonin 5-HT1D receptor mRNA in mouse oocytes and preimplantation embryos was confirmed by in situ hybridization showing a clearly distinct punctate signal. Immunocytochemistry results revealed the localization of serotonin in oocytes and embryos to the blastocyst stage as diffuse punctate cytoplasmic labelling. It appears that endogenous and/or exogenous serotonin in preimplantation embryos could be involved in complex autocrine/paracrine regulations of embryo development and embryo-maternal interactions.


2006 ◽  
Vol 27 (1) ◽  
pp. 352-367 ◽  
Author(s):  
Daisuke Mori ◽  
Yoshihisa Yano ◽  
Kazuhito Toyo-oka ◽  
Noriyuki Yoshida ◽  
Masami Yamada ◽  
...  

ABSTRACT NDEL1 is a binding partner of LIS1 that participates in the regulation of cytoplasmic dynein function and microtubule organization during mitotic cell division and neuronal migration. NDEL1 preferentially localizes to the centrosome and is a likely target for cell cycle-activated kinases, including CDK1. In particular, NDEL1 phosphorylation by CDK1 facilitates katanin p60 recruitment to the centrosome and triggers microtubule remodeling. Here, we show that Aurora-A phosphorylates NDEL1 at Ser251 at the beginning of mitotic entry. Interestingly, NDEL1 phosphorylated by Aurora-A was rapidly downregulated thereafter by ubiquitination-mediated protein degradation. In addition, NDEL1 is required for centrosome targeting of TACC3 through the interaction with TACC3. The expression of Aurora-A phosphorylation-mimetic mutants of NDEL1 efficiently rescued the defects of centrosomal maturation and separation which are characteristic of Aurora-A-depleted cells. Our findings suggest that Aurora-A-mediated phosphorylation of NDEL1 is essential for centrosomal separation and centrosomal maturation and for mitotic entry.


Development ◽  
1993 ◽  
Vol 117 (1) ◽  
pp. 341-345 ◽  
Author(s):  
P.S. Burgoyne

Karyotopic and cell number analysis of 3.5 day post coitum preimplantation mouse embryos was used to determine whether XY embryos had more cells than XX embryos at the late morula/early blastocyst stage. This proved to be the case for the CD1 strain (for which it had previously been shown that XY embryos form a blastocoel earlier than XX embryos) and for the MF1 strain. However, this increased cell number was not seen in MF1 embryos carrying an RIII strain Y in place of the MF1 Y. Furthermore, interstrain crosses between CD1 and the MF1, YRIII strain showed that the cell number increase segregated with the CD1 Y but not with the RIII Y. It is concluded that the CD1 and MF1 Y chromosomes carry a factor that accelerates the rate of preimplantation development.


2021 ◽  
Vol 33 (12) ◽  
pp. 725
Author(s):  
Dawid Winiarczyk ◽  
Anna Piliszek ◽  
Silvestre Sampino ◽  
Marek Lukaszewicz ◽  
Jacek Andrzej Modliński

Programmed cell death plays a key role in mammalian development because the morphological events of an organism’s formation are dependent on apoptosis. In the mouse development, the first apoptotic waves occur physiologically at the blastocyst stage. Cell number and the mean nucleus to cytoplasm (N/C) ratio increase exponentially throughout subsequent embryo cleavages, while cell volume concurrently decreases from the zygote to blastocyst stage. In this study we tested the hypothesis that reorganisation of the embryo structure by manipulating cell number, the N/C ratio and the cell volume of 2-cell embryos may result in the earlier and more frequent occurrence of apoptosis. The results indicate that doubling (‘Aggregates’ group) or halving (‘Embryos 1/2’ group) the initial cell number and modifying embryo volume, ploidy (‘Embryos 4n’ group) and the N/C ratio (‘Embryos 2/1’ group) reduce the probability of apoptosis in the resulting embryos. There was a higher probability of apoptosis in the inner cell mass of the blastocyst, but apoptotic cells were never observed at the morula stage in any of the experimental groups. Thus, manipulation of cell number, embryo volume, the N/C ratio and ploidy cause subtle changes in the occurrence of apoptosis, although these are mostly dependent on embryo stage and cell lineage (trophectoderm or inner cell mass), which have the greatest effect on the probability of apoptosis.


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
L Bori ◽  
M.Á Valera ◽  
D Gilboa ◽  
R Maor ◽  
I Kottel ◽  
...  

Abstract Study question Can we distinguish between top-grade euploid and aneuploid embryos by AI measurement of cell edges in time-lapse videos? Summary answer Aneuploid embryos can be distinguished from euploid embryos by AI determination of a longer time to blastulation and higher cell activity. What is known already Continuous monitoring of the embryo development has brought out morphokinetic parameters that are used to predict pre-implantation genetic testing (PGT) results. Previous publications showed that euploid embryos reach blastulation earlier than non-euploid embryos. However, time-lapse data are currently under-utilized in making predictions about embryo chromosomal content. AI and computer vision could take advantage of the massive amount of data embedded in the images of embryo development. This is the first attempt to distinguish between euploid and aneuploid embryos by computer vision in an objective and indirect way based on the measurement of cell edges as a proxy for cell activity. Study design, size, duration We performed a retrospective analysis of 1,314 time-lapse videos from embryos cultured to the blastocyst stage with PGT results. This single-center study involved two phases; a comparison of the start time of blastulation between euploid (n = 544) and aneuploid embryos (n = 797). In phase two, we designed a novel methodology to examine whether precise measurement of cell edges over time could reflect cell activity differences in blastulation. Participants/materials, setting, methods We assumed that the delay in blastulation is reflected by higher cell activity that could be determined accurately for the first time using computer vision and machine learning to measure the length of the edges (from t2 to t8). We compared computer vision based measurements of cell edges, reflecting cell number and size, in videos of 231 top-grade euploid (n = 111) and aneuploid (n = 120) embryos. Main results and the role of chance The mean and standard deviation of blastulation start time was 100.1±6.8 h for euploid embryos and 101.8±8.2 h for aneuploid embryos (p < 0.001). Regarding the measurement of cell activity, a computer vision algorithm identified the edges and provided a certainty score for each edge, higher when the algorithm is more certain that this is a cell edge (as opposed to noise in the images). A threshold was set to distinguish cell edges from noise using this score. The following results for top-grade embryos are shown as the sum of the edge lengths (µm) average of 160 pictures per embryo (frames between t2 and t8). The total length of the cell edges increased from two cells (420±85 µm) to eight cells (861±237 µm), in line with the mitosis events. Both the average total edge measured (450±162 µm for euploid embryos and 489±215 µm for aneuploid embryos, p < 0.01) and the average total of the difference between consecutive frames (135±47 µm for euploid embryos and 153±64 µm for aneuploid embryos, p < 0.01) were higher for aneuploid embryos than for euploid embryos. A regression model to differentiate between the two classes achieved 73% sensitivity and 73% specificity on this dataset. Limitations, reasons for caution The main limitation of this study is the difficulty to correlate our findings to other measure of cell activity. A more robust AI function (using not only cell edges lengths) would be required for future analysis to measure the cell activity in cell division up to the blastocyst stage. Wider implications of the findings Our results show for the first time that an AI based system can precisely measure microscopic cell edges in the dividing embryo. Using this novel method, we could distinguish between euploid and aneuploid embryos. This non-invasive method could further enhance our knowledge of the developing embryo. Trial registration number Not Applicable


Development ◽  
1991 ◽  
Vol 113 (Supplement_2) ◽  
pp. 157-168 ◽  
Author(s):  
Gary C. Schoenwolf

Neuruiation, formation of the neural tube, a crucial event of early embryogenesis, is believed to be driven by the coordination of a number of diverse morphogenetic cell behaviors. Such behaviors include changes in cell number (division, death), cell shape and size (wedging, palisading and spreading), cell position (rearrangement or intercalation) and cell–cell and cell–matrix associations (including inductive interactions). The focus of this essay is on epiblast cell movements and their role in shaping and bending of the neural plate. Neuruiation is a multifactorial process requiring both intrinsic (within the neural plate) and extrinsic (outside the neural plate) forces. The origin and movements of three populations of epiblast cells have been studied in avian embryos by constructing quail/chick transplantation chimeras and by labeling cells in situ with identifiable, heritable markers. MHP (median hinge-point neurepithelial) cells originate principally from a midline epiblast area rostral to and overlapping Hensen's node. In addition, a few caudal MHP cells originate from paranodal epiblast areas. MHP cells stream down the length of the midline neuraxis in the wake of the regressing Hensen's node. This streaming occurs as a result of cell division (presumably oriented so that daughter cells are placed into the longitudinal plane rather than into the transverse plane) and rearrangement (intercalation), resulting in a narrowing of the width of the MHP region with a concomitant increase in its length. L (lateral neurepithelial) cells originate from paired epiblast areas flanking the rostral portion of the primitive streak, and they stream down the length of the lateral neuraxis concomitant with regression of Hensen's node. They do so both by oriented cell division and by intercalation. SE (surface epithelial) cells originate from the epiblast of the area pellucida, as far lateral as near the area pellucida area opaca border. From this area they stream medially, toward the forming lateral margins of the neural plate. Collectively, movements of the three populations of epiblast cells generate the convergent-extension movements characteristic of the epiblast during neuruiation. Heterotopic grafting has been used to assess the relationship between cell position and cell fate and to determine whether transplanted heterotopic cells can adopt the behaviors typical of the new site. For example, SE cells can replace L cells, changing their fate and adopting L-cell behavior. Similarly, prospective MHP and L cells both can change their fate and adopt the behavior of SE cells. L cells, when placed into prospective MHP-cell territory, move out of this territory by intermingling with adjacent host L cells. Likewise, prospective MHP cells placed into L-cell territory, move out of this territory by intermingling with host MHP cells. Collectively, these results suggest that cell fate is determined principally by the ultimate position of cells, and that adjacent, different cell populations are restricted from intermingling with one another. How positional information is specified, the nature of restriction of intermingling and the guidance cues used for cell navigation during streaming remain to be elucidated.


2002 ◽  
Vol 172 (2) ◽  
pp. 221-236 ◽  
Author(s):  
K Hardy ◽  
S Spanos

There is increasing evidence that even before implantation, human development is regulated by embryonically and maternally derived growth factors. Studies in other mammalian species have shown that growth factors and their receptors are expressed by the preimplantation embryo and the reproductive tract. Furthermore, a number of growth factors have been shown to affect rate of embryo development, the proportion of embryos developing to the blastocyst stage, blastocyst cell number, metabolism and apoptosis. Growth factor ligands and receptors are also expressed in human embryos and the maternal reproductive tract, and supplementation of culture medium with exogenous growth factors affects cell fate, development and metabolism of human embryos in vitro. Autocrine, paracrine and endocrine pathways that may operate within the embryo and between the embryo and the reproductive tract before implantation are proposed.


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