Asynchronous Cell Division Research Articles

Changes in repair pathways of radiation-induced DNA double-strand breaks at the midblastula transition in Xenopus embryo.

Ionizing radiation (IR) causes DNA damage, particularly DNA double-strand breaks (DSBs), which have significant implications for genome stability. The major pathways of repairing DSBs are homologous recombination (HR) and nonhomologous end joining (NHEJ). However, the repair mechanism of IR-induced DSBs in embryos is not well understood, despite extensive research in somatic cells. The externally developing aquatic organism, Xenopus tropicalis, serves as a valuable model for studying embryo development. A significant increase in zygotic transcription occurs at the midblastula transition (MBT), resulting in a longer cell cycle and asynchronous cell divisions. This study examines the impact of X-ray irradiation on Xenopus embryos before and after the MBT. The findings reveal a heightened X-ray sensitivity in embryos prior to the MBT, indicating a distinct shift in the DNA repair pathway during embryo development. Importantly, we show a transition in the dominant DSB repair pathway from NHEJ to HR before and after the MBT. These results suggest that the MBT plays a crucial role in altering DSB repair mechanisms, thereby influencing the IR sensitivity of developing embryos.

Toxicity of Orthodontic Brackets Examined by Single Cell Tracking.

Subtle toxic effects may be masked in traditional assays that average or summate the response of thousands of cells. We overcome this by using the recent method of single cell tracking in time-lapse recordings. This follows the fate and behavior of individual cells and their progeny and provides unambiguous results for multiple simultaneous biological responses. Further, single cell tracking permits correlation between progeny relationships and cell behavior that is not otherwise possible, including disruption by toxins and toxicants of similarity between paired sister cells. Notably, single cell tracking seems not to have been previously used to study biomaterials toxicity. The culture medium was pre-conditioned by 79 days incubation with orthodontic brackets from seven separate commercial sources. Metal levels were determined by Inductively Coupled Plasma Mass Spectrometry. Metal levels varied amongst conditioned media, with elevated Cr, Mn, Ni, and Cu and often Mo, Pb, Zn, Pd, and Ag were occasionally found. The effect on human dermal fibroblasts was determined by single cell tracking. All bracket-conditioned media reduced cell division (p < 0.05), while some reduced cell migration (p < 0.05). Most bracket-conditioned media increased the rate of asynchronous sister cell division (p < 0.05), a seemingly novel measure for toxicity. No clear effect on cell morphology was seen. We conclude that orthodontic brackets have cytotoxic effects, and that single cell tracking is effective for the study of subtle biomaterials cytotoxicity.

MiR-202-3p determines embryo viability during mid-blastula transition.

Developmental growth is an intricate process involving the coordinated regulation of the expression of various genes, and microRNAs (miRNAs) play crucial roles in diverse processes throughout animal development. The mid-blastula transition (MBT) is a developmental milestone when maternal RNAs are cleared and the zygotic genome programmed asynchronous cell division begins to drive embryogenesis. While mechanisms underlying MBT have been intensively revealed, factors regulating cell proliferation at the transition remain largely unknown. We report here a microRNA, miR-202-3p to be a key factor that determines embryonic fate during MBT in zebrafish. A miR-202-3p antagomir specifically terminated embryo development at the mid-blastula stage. In vivo deletion of the miR-202 locus recapitulated the fatal phenotypes, which were rescued only by miR-202-3p or its precursor. Transcriptome comparison revealed >250 RNAs including both maternal and zygotic origins were dysregulated at MBT in the miR-202−/− embryos, corresponding with arrays of homeostatic disorders leading to massive apoptosis. A trio of genes: nfkbiaa, perp and mgll, known to be intimately involved with cell proliferation and survival, were identified as direct targets of miR-202-3p. Importantly, over- or under-expression of any of the trio led to developmental delay or termination at the blastula or gastrula stages. Furthermore, nfkbiaa and perp were shown to inter-regulate each other. Thus, miR-202-3p mediates a regulatory network whose components interact closely during MBT to determine embryonic viability and development.

P-136 Cell exclusion during human embryo development results in altered morphokinetic patterns up to morula formation

Abstract Study question Do embryos that present with cell exclusion show dysregulated cleavage divisions and deviating reproductive success, compared to normally dividing embryos? Summary answer Exclusion embryos have lower reproductive success, showing patterns of delayed and rapid cleavage until morulation, suggesting potential compensatory mechanisms and cellular checkpoints in post-compaction stages What is known already Within assisted reproductive technology, visual observation of embryo development constitutes a major contribution to assessments of embryo competence. Time-lapse technology has allowed for detailed identification and measurement of dynamic events that impact embryo development and reproductive outcome. Two recent studies have identified an anomalous pattern of cell division occurring during early embryo development, termed cell exclusion. These cells seem to arrest during early cleavage stages and fail to participate in downstream compaction, thereby resulting in lower reproductive success. Previous research suggests aberrant chromosomal re-arrangements or irregular cellular content may contribute to negatively associated reproductive outcomes. Study design, size, duration This study is based on a single-clinic retrospective analysis of 515 time-lapse videos collected from 2013-2019. Embryos were cultured in EmbryoScope incubators for 4-5 days post-insemination by conventional in vitro fertilization or intracytoplasmic sperm injection, followed by fresh transfer. Supervised annotations were made for cell divisions and morphological quality by three embryologists. An in-depth morphokinetic analysis was conducted using student’s t-test and analysis of variance for continuous variables. Participants/materials, setting, methods A total of 515 embryo videos and associated annotations were obtained from 502 patients. All videos were individually assessed and grouped according to the presence (n = 122) or absence (n = 393) of an excluded cell. Cell division annotations were collected from the 2-cell stage to blastocyst stage for each video. Time annotations and time intervals between subsequent cell divisions were analyzed to determine differences, if any, between control and exclusion embryos. Main results and the role of chance Following transfer, exclusion embryos resulted in significantly lower rates of fetal heartbeat (28% vs 48%; p &amp;lt; 0.05) and live birth (20% vs 40%; p &amp;lt; 0.05), compared to control embryos. A strong negative association was found between cell exclusion and reproductive success (OR = 0.41; 95% CI = 0.27-0.64; p &amp;lt; 0.0001). Time annotations for 2-cell, 4-cell, and 6-cell to 9-cell stages in the exclusion group showed delayed cell divisions by an average of 3.16 hours up to morulation, compared to control embryos (p &amp;lt; 0.05). However, times to 3-cell and 5-cell stages were significantly earlier than control embryos (p = 0.03 and p = 0.0002, respectively). Exclusion embryos seemed to spend more time in “intermediate” cell stages such as the 3-cell, 5-cell and 6-cell stages (p &amp;lt; 0.01), compared to control embryos. Conversely, they spent less time in commonly known “checkpoint” stages such as the 2-cell, 4-cell, and 8-cell stage (p &amp;lt; 0.001). Moreover, exclusion embryos were found to be more asynchronous than control embryos (p &amp;lt; 0.0001) despite similar cell cycle lengths. No significant morphokinetic differences were found from the start of blastulation to full expansion before zona thinning, between experimental groups. Subgroup analysis revealed no significant trends for known confounding factors pertaining to cell division such as maternal age, paternal age, maternal body mass index and infertility diagnosis. Limitations, reasons for caution Aside from this study being retrospective, there may be unknown confounders that could impact this event. Moreover, identification of cell exclusion was conducted in a single data set generated by one clinic and was only performed by one person, thus limiting the generalizability of the findings. Wider implications of the findings Our results indicate that cell exclusion has negative associations with reproductive outcome. Moreover, morphokinetic analysis has been able to identify aspects of asynchronous and dysregulated cell division that are associated with cell exclusion. Further analysis combining morphokinetics with genetic testing and adhesion protein investigation may improve current embryo assessments. Trial registration number N/A

Ultrastructure and in-situ chemical characterization of intracellular granules of embryo-like fossils from the early Ediacaran Weng’an biota

Embryo-like fossils from the early Ediacaran Weng’an biota provide a window of exceptional fossil preservation onto the period of life history in which molecular clocks estimate the fundamental animal lineages to have diverged. However, their diversity and biological affinities have proven controversial, because they are morphologically simple and, consequently, their interpretation lacks phylogenetic constraint. The subcellular structures preserved in these embryo-like fossils might help to understand their cytology, biology, and diversity, but the potential of these structures has not been fully realized, because detailed microscale physical and chemical investigations are lacking. Here, to remedy this deficiency, we performed a comprehensive study to characterize their micro- and ultra-structures as well as in-situ chemical components. Our results reveal three types of subcellular structure that differ in size, shape, and mineral components: (1) relatively small and spheroidal granules in embryo-like fossils with equal cell division pattern; (2) relatively large, spheroidal, or polygonal granules in embryo-like fossils with unequal and asynchronous cell division pattern; and (3) irregular multi-layered rim-bounded granules in embryo-like fossils with unequal and asynchronous cell division pattern. We propose that the three types may be rationalized to a single taphonomic pathway of preferential mineralization of the cell cytoplasm, preserving an external mould of subcellular granules. We followed the previous interpretation that the spheroidal and polygonal granules should be fossilized lipid droplets or yolk platelets. The distinction between these subcellular structures are largely the result of postmortem degradation processes such as autolysis. The widely preserved lipid droplets or yolk platelets within these Ediacaran embryo-like fossils are compatible with the interpretion of large yolky embryos with maternal nourishment and direct development.

Karyotype Reorganization in Wheat-Rye Hybrids Obtained via Unreduced Gametes: Is There a Limit to the Chromosome Number in Triticale?

To date, few data have been accumulated on the contribution of meiotic restitution to the formation of Triticum aestivum hybrid karyotypes. In this study, based on FISH and C-banding, karyotype reorganization was observed in three groups of F5 wheat–rye hybrids 1R(1A) × R. Aberrations, including aneuploidy, telocentrics, and Robertsonian translocations, were detected in all groups. Some of the Group 1 plants and all of the Group 2 plants only had a 4R4R pair (in addition to 1R1R), which was either added or substituted for its homeolog in ABD subgenomes. In about 82% of meiocytes, 4R4R formed bivalents, which indicates its competitiveness. The rest of the Group 1 plants had 2R and 7R chromosomes in addition to 1R1R. Group 3 retained all their rye chromosomes, with a small aneuploidy on the wheat chromosomes. A feature of the meiosis in the Group 3 plants was asynchronous cell division and omission of the second division. Diploid gametes did not form because of the significant disturbances during gametogenesis. As a result, the frequency of occurrence of the formed dyads was negatively correlated (r = −0.73) with the seed sets. Thus, meiotic restitution in the 8n triticale does not contribute to fertility or increased ploidy in subsequent generations.

Evaluation of dimethoate toxicity on fertilization and on embryonic development of Paracentrotus lividus (Lamarck, 1816).

Organophosphates are a large class of chemicals with anticholinesterase action insecticides. Dimethoate belongs to the class of organophosphates and it is used for agriculture purpose. Its main toxicological role in animals and humans is the inhibition of the activity of acetylcholinesterase. Although it is not considered genotoxic, carcinogenic and teratogen, there is evidence of increased pup mortality in developmental neurotoxicity studies. Since there is scant published literature about developmental toxicity, we investigated the adverse effects of dimethoate on fertilization and embryonic development in sea urchin (Paracentrotus lividus), a model organism widely used to assess the toxicity of contaminants on environmental matrices; so pesticide residues can be released into the environment, and could affect the health of organisms, including humans. Different solution of dimethoate (4×10-3, 4×10-4, 4×10-5, 4×10-6 and 4×10-7g/10ml) have been tested on spermatozoa of P. lividus to evaluate the fertilizing ability of them when we added egg cells untreated. We demonstrated that dimethoate does not interfere with fertilizing ability of spermatozoa but egg cells fertilized by treated spermatozoa showed alterations in the segmentation planes as asymmetric and/or asynchronous cell divisions.

Mono- to Multilayer Transition in Growing Bacterial Colonies.

The transition from monolayers to multilayered structures in bacterial colonies is a fundamental step in biofilm development. Observed across different morphotypes and species, this transition is triggered within freely growing bacterial microcolonies comprising a few hundred cells. Using a combination of numerical simulations and analytical modeling, here we demonstrate that this transition originates from the competition between growth-induced in-plane active stresses and vertical restoring forces, due to the cell-substrate interactions. Using a simple chainlike colony of laterally confined cells, we show that the transition sets when individual cells become unstable to rotations; thus it is localized and mechanically deterministic. Asynchronous cell division renders the process stochastic, so that all the critical parameters that control the onset of the transition are continuously distributed random variables. Here we demonstrate that the occurrence of the first division in the colony can be approximated as a Poisson process in the limit of large cell numbers. This allows us to approximately calculate the probability distribution function of the position and time associated with the first extrusion. The rate of such a Poisson process can be identified as the order parameter of the transition, thus highlighting its mixed deterministic-stochastic nature.

Effect of magnetic fields on the growth of bacterium Staphylococcus aureus

The effects of weak static and low-frequency magnetic fields on the growth rate of S. aureus were investigated. The measurements were performed with static magnetic fields with magnitude in range 0.0 G ≤ Ho ≤ 14.0 G and low-frequency magnetic fields at fixed amplitude Ho 4.5 G, and frequency range 0.0 ≤ f ≤ 1.0 kHz. The growth of these bacteria is negatively affected by increasing the intensity of the static magnetic field. When exposed to an oscillating field, a positive effect was observed on the rate of growth of the colonies with respect to the field frequency. In both, static or ac magnetic field, the growth curves follow an exponential law, typical of asynchronous cell divisions.

Selection for synchronized cell division in simple multicellular organisms

The evolution of multicellularity was a major transition in the history of life on earth. Conditions under which multicellularity is favored have been studied theoretically and experimentally. But since the construction of a multicellular organism requires multiple rounds of cell division, a natural question is whether these cell divisions should be synchronous or not. We study a population model in which there compete simple multicellular organisms that grow by either synchronous or asynchronous cell divisions. We demonstrate that natural selection can act differently on synchronous and asynchronous cell division, and we offer intuition for why these phenotypes are generally not neutral variants of each other.

Morphogenetic movements affect local tissue organisation during embryonic Drosophila morphogenesis

Tissues and organs undergo extensive remodelling to reach their final morphology and physiological activity. The genetic programs underlying tissue formation are well studied, but less is known about how this formation is influenced by extrinsic forces derived from other concomitant morphogenetic events. Here we address this question in Drosophila melanogaster. We analyse tissue organisation in the embryonic epidermis at stage 10 by computational tissue segmentation methods to provide a quantitative description of packing. We find that the epidermis adopts different organisations along the dorso-ventral axis that correlate with differences in cell density. We analyse the contribution of three morphogenetic events that take place right before or concomitant to this period of embryogenesis, neuroblast delamination, asynchronous postblastoderm cell divisions and germ band extension, and we find that they all exert an influence on the packing of the epidermis. We previously described that the apical determinant Crumbs accumulates differentially in the epidermis along the dorso-ventral axis. Here we find that this differential accumulation of Crumbs correlates with the differential tissue packing. Perturbation of the three mentioned morphogenetic events also modulate Crumbs differential accumulation, suggesting that Crb could act as a read-out of tissue organisation. We also previously showed that Crb plays a role in regulating cell architecture. Now we find that it is also required for proper packing of the embryonic epidermis. In summary, here we uncover an intimate relationship between morphogenetic events and cell packing within a tissue that is dependent on surrounding cell density. Furthermore we find that this morphogenetically-regulated tissue packing modulates the key cell polarity protein Crumbs, which in turn is required for tissue packing, suggesting that it may participate in the molecular mechanism/s underlying the described tissue organisation.

The invariant cleavage pattern displayed by ascidian embryos depends on spindle positioning along the cell's longest axis in the apical plane and relies on asynchronous cell divisions.

The ascidian embryo is an ideal system to investigate how cell position is determined during embryogenesis. Using 3D timelapse imaging and computational methods we analyzed the planar cell divisions in ascidian early embryos and found that spindles in every cell tend to align at metaphase in the long length of the apical surface except in cells undergoing unequal cleavage. Furthermore, the invariant and conserved cleavage pattern of ascidian embryos was found to consist in alternate planar cell divisions between ectoderm and endomesoderm. In order to test the importance of alternate cell divisions we manipulated zygotic transcription induced by β-catenin or downregulated wee1 activity, both of which abolish this cell cycle asynchrony. Crucially, abolishing cell cycle asynchrony consistently disrupted the spindle orienting mechanism underpinning the invariant cleavage pattern. Our results demonstrate how an evolutionary conserved cell cycle asynchrony maintains the invariant cleavage pattern driving morphogenesis of the ascidian blastula.

Antagonistic regulation of the second mitotic wave by Eyes absent-Sine oculis and Combgap coordinates proliferation and specification in the Drosophila retina.

The transition from proliferation to specification is fundamental to the development of appropriately patterned tissues. In the developing Drosophila eye, Eyes absent (Eya) and Sine oculis (So) orchestrate the progression of progenitor cells from asynchronous cell division to G1 arrest and neuronal specification at the morphogenetic furrow. Here, we uncover a novel role for Eya and So in promoting cell cycle exit in the second mitotic wave (SMW), a synchronized, terminal cell division that occurs several hours after passage of the furrow. We show that Combgap (Cg), a zinc-finger transcription factor, antagonizes Eya-So function in the SMW. Based on the ability of Cg to attenuate Eya-So transcriptional output in vivo and in cultured cells and on meta analysis of their chromatin occupancy profiles, we speculate that Cg limits Eya-So activation of select target genes posterior to the furrow to ensure properly timed mitotic exit. Our work supports a model in which context-specific modulation of transcriptional activity enables Eya and So to promote both entry into and exit from the cell cycle in a distinct spatiotemporal sequence.

Comparative cytological and transcriptomic analysis of pollen development in autotetraploid and diploid rice

Autotetraploid rice has greater genetic variation and higher vigor than diploid rice, but low pollen fertility is one of the major reasons for low yield of autotetraploid rice. Very little is known about the molecular mechanisms of low pollen fertility of autotetraploid rice. In this study, cytological observations and microarray analysis were used to assess the genetic variation during pollen development in autotetraploid and diploid rice. Many abnormal chromosome behaviors, such as mutivalents, lagged chromosomes, asynchronous cell division, and so on, were found during meiosis in autotetraploid. Microsporogenesis and microgametogenesis in autotetraploid rice was similar to diploid rice, but many different kinds of abnormalities, including microspores degeneration, multi-aperture, and abnormal cell walls, were found in autotetraploid rice. Compared with diploid rice, a total of 1,251 genes were differentially expressed in autotetraploid rice in pollen transcriptome, among them 1,011 and 240 genes were up-regulated and down-regulated, respectively. 124 and 6 genes were co-up-regulated and co-down-regulated during three pollen development stages, respectively. These results suggest that polyploidy induced up-regulation for most of the genes during pollen development. Quantitative RT-PCR was done to validate 12 differentially expressed genes selected from functional categories based on the gene ontology analysis. These stably expressed genes not only related to the pollen development genes, but also involved in cell metabolism, cell physiology, binding, catalytic activity, molecular transducer activity, and transcription regulator activity. The present study suggests that differential expression of some key genes may lead to complex gene regulation and abnormal pollen development in autotetraploid rice.

PAR-4/LKB1 regulates DNA replication during asynchronous division of the early C. elegans embryo.

Regulation of cell cycle duration is critical during development, yet the underlying molecular mechanisms are still poorly understood. The two-cell stage Caenorhabditis elegans embryo divides asynchronously and thus provides a powerful context in which to study regulation of cell cycle timing during development. Using genetic analysis and high-resolution imaging, we found that deoxyribonucleic acid (DNA) replication is asymmetrically regulated in the two-cell stage embryo and that the PAR-4 and PAR-1 polarity proteins dampen DNA replication dynamics specifically in the posterior blastomere, independently of regulators previously implicated in the control of cell cycle timing. Our results demonstrate that accurate control of DNA replication is crucial during C. elegans early embryonic development and further provide a novel mechanism by which PAR proteins control cell cycle progression during asynchronous cell division.

MAP Kinase Signaling Antagonizes PAR-1 Function During Polarization of the Early Caenorhabditis elegans Embryo

PAR proteins (partitioning defective) are major regulators of cell polarity and asymmetric cell division. One of the par genes, par-1, encodes a Ser/Thr kinase that is conserved from yeast to mammals. In Caenorhabditis elegans, par-1 governs asymmetric cell division by ensuring the polar distribution of cell fate determinants. However the precise mechanisms by which PAR-1 regulates asymmetric cell division in C. elegans remain to be elucidated. We performed a genomewide RNAi screen and identified six genes that specifically suppress the embryonic lethal phenotype associated with mutations in par-1. One of these suppressors is mpk-1, the C. elegans homolog of the conserved mitogen activated protein (MAP) kinase ERK. Loss of function of mpk-1 restored embryonic viability, asynchronous cell divisions, the asymmetric distribution of cell fate specification markers, and the distribution of PAR-1 protein in par-1 mutant embryos, indicating that this genetic interaction is functionally relevant for embryonic development. Furthermore, disrupting the function of other components of the MAPK signaling pathway resulted in suppression of par-1 embryonic lethality. Our data therefore indicates that MAP kinase signaling antagonizes PAR-1 signaling during early C. elegans embryonic polarization.

Reconstructing the ancestral female gametophyte of angiosperms: Insights from Amborella and other ancient lineages of flowering plants

For more than a century, the common ancestor of flowering plants was thought to have had a seven-celled, eight-nucleate Polygonum-type female gametophyte. It is now evident that not one, but in fact three, patterns of female gametophyte development and mature structure characterize the common ancestors of the four most ancient clades of extant angiosperms: Amborella-type, Nuphar/Schisandra-type and Polygonum-type. The Amborella-type female gametophyte is restricted to a single extant species, Amborella trichopoda, and at maturity consists of eight cells and nine nuclei. Development of the Amborella-type gametophyte is essentially identical to the Polygonum-type except that there is an additional and asynchronous cell division at the micropylar pole prior to maturation that produces a third synergid and the egg cell. The Nuphar/Schisandra-type female gametophyte is four-nucleate and four-celled and at maturity contains a typical three-celled egg apparatus and a central cell with a single haploid polar nucleus. This type of gametophyte appears to be universal among extant members of the Nymphaeales (including Hydatellaceae) and Austrobaileyales. Based on explicit reconstruction of character distribution and evolution, the Polygonum-type female gametophyte is certain to be representative of the common ancestors of monocots, eudicots, magnoliids, Ceratophyllaceae, and Chloranthaceae. There are compelling biological reasons to suggest that the four-celled, four-nucleate female gametophyte (as found in Nymphaeales and Austrobaileyales) is ancestral among angiosperms, with transitions to Polygonum-type female gametophytes separately in the Amborellales and in the ancient angiosperm clade that includes all angiosperms except Amborella, Nymphaeales, and Austrobaileyales. Subsequent to the evolution of a seven-celled, eight-nucleate Polygonum-type female gametophyte in the Amborellales, we hypothesize that a peramorphic increase in egg apparatus cell number took place and led to the unique situation in which there are three synergids in Amborella trichopoda.

Maternal cyclin B levels “Chk” the onset of DNA replication checkpoint control in Drosophila

In many animals, early development of the embryo is characterized by synchronous, biphasic cell divisions. These cell divisions are controlled by maternally inherited proteins and RNAs. A critical question in developmental biology is how the embryo transitions to a later pattern of asynchronous cell divisions and transfers the prior maternal control of development to the zygotic genome. The most-common model regarding how this transition from maternal to zygotic control is regulated posits that this is a consequence of the limitation of maternal gene products, due to their titration during early cell divisions. Here we discuss a recent article by Crest et al.1 that instead proposes that the balance of Cyclin-dependent Kinase 1 and Cyclin B (Cdk1-CycB) activity relative to that of the Drosophila checkpoint kinase Chk1 determines when asynchronous divisions begin.

Mathematical modeling of cell population dynamics in the colonic crypt and in colorectal cancer

Colorectal cancer is initiated in colonic crypts. A succession of genetic mutations or epigenetic changes can lead to homeostasis in the crypt being overcome, and subsequent unbounded growth. We consider the dynamics of a single colorectal crypt by using a compartmental approach [Tomlinson IPM, Bodmer WF (1995) Proc Natl Acad Sci USA 92:], which accounts for populations of stem cells, differentiated cells, and transit cells. That original model made the simplifying assumptions that each cell population divides synchronously, but we relax these assumptions by adopting an age-structured approach that models asynchronous cell division, and by using a continuum model. We discuss two mechanisms that could regulate the growth of cell numbers and maintain the equilibrium that is normally observed in the crypt. The first will always maintain an equilibrium for all parameter values, whereas the second can allow unbounded proliferation if the net per capita growth rates are large enough. Results show that an increase in cell renewal, which is equivalent to a failure of programmed cell death or of differentiation, can lead to the growth of cancers. The second model can be used to explain the long lag phases in tumor growth, during which new, higher equilibria are reached, before unlimited growth in cell numbers ensues.

Cellular and Subcellular Structure of Neoproterozoic Animal Embryos

Stereoblastic embryos from the Doushantuo Formation of China exhibit occasional asynchronous cell division, with diminishing blastomere volume as cleavage proceeded. Asynchronous cell division is common in modern embryos, implying that sophisticated mechanisms for differential cell division timing and embryonic cell lineage differentiation evolved before 551 million years ago. Subcellular structures akin to organelles, coated yolk granules, or lipid vesicles occur in these embryos. Paired reniform structures within embryo cells may represent fossil evidence of cells about to undergo division. Embryos exhibit no evidence of epithelial organization, even in embryos composed of approximately 1000 cells. Many of these features are compatible with metazoans, but the absence of epithelialization is consistent only with a stem-metazoan affinity for Doushantuo embryos.