Embryonic Cell Division Research Articles

RETRACTED: Fam208a orchestrates interaction protein network essential for early embryonic development and cell division

Maintenance of genome stability is essential for every living cell as genetic information is repeatedly challenged during DNA replication in each cell division event. Errors, defects, delays, and mistakes that arise during mitosis or meiosis lead to an activation of DNA repair processes and in case of their failure, programmed cell death, i.e. apoptosis, could be initiated. Fam208a is a protein whose importance in heterochromatin maintenance has been described recently. In this work, we describe the crucial role of Fam208a in sustaining the genome stability during the cellular division. The targeted depletion of Fam208a in mice using CRISPR/Cas9 leads to embryonic lethality before E12.5. We also used the siRNA approach to downregulate Fam208a in zygotes to avoid the influence of maternal RNA in the early stages of development. This early downregulation increased arresting of the embryonal development at the two-cell stage and occurrence of multipolar spindles formation. To investigate this further, we used the yeast two-hybrid (Y2H) system and identified new putative interaction partners Gpsm2, Amn1, Eml1, Svil, and Itgb3bp. Their co-expression with Fam208a was assessed by qRT-PCR profiling and in situ hybridisation [1] in multiple murine tissues. Based on these results we proposed that Fam208a functions within the HUSH complex by interaction with Mphosph8 as these proteins are not only able to physically interact but also co-localise. We are bringing new evidence that Fam208a is multi-interacting protein affecting genome stability on the level of cell division at the earliest stages of development and also by interaction with methylation complex in adult tissues. In addition to its epigenetic functions, Fam208a appears to have an additional role in zygotic division, possibly via interaction with newly identified putative partners Gpsm2, Amn1, Eml1, Svil, and Itgb3bp.

Proteomics of bovine endometrium, oocytes and early embryos

Summary. Embryo-maternal communication, as well as recognition and establishment of pregnancy, is predominantly controlled by secretion of hormones and proteins. Furthermore, uterine fluid serves as a histotroph and contains factors essential for embryo development and elongation, making it a meaningful source for proteomic approaches of early embryonic development. The studies included in this review address the analysis of uterine fluid and endometrium during the oestrous cycle, early pregnancy, maternal effects on embryo development and embryo-induced alterations to the maternal environment. Clear differences between the proteome of uterine fluid and blood plasma unravelled the active contribution of the endometrium to the composition of uterine luminal fluid. Proteins were identified which may contribute either to growth enhancement of blastocysts or influence secretion of "Interferon tau". Several studies suggested important roles of intrauterine factors including "Tissue inhibitor of metalloproteinases 2" for pregnancy establishment and "Acyl-CoA-binding protein" for the embryo prior to implantation. A meta-analysis of quantitative proteome studies revealed functional clusters including the terms "degradation of extracellular matrix", "platelet degranulation" and "MHC class II antigen presentation". Proteomic studies targeting oocytes and early embryos provided evidence for the function of "Cyclin B" in mitotic resumption, for "14-3-3 epsilon" to activate "maturation promotion factor" for promotion of mitosis during in vitro maturation, or for the inhibition of proteasomes during the late phase of maturation. Analysis of blastocyst derived trophectoderm cell lines suggested effects of annexin down-regulation on the failure rate of nuclear transfer pregnancies. Finally, absolute quantification of "Y-box-binding protein 2" and "Insulin-like growth factor 2 mRNA-binding protein 3" during early embryogenesis by "selected reaction monitoring" demonstrated the dynamism of proteome alterations during the first embryonic cell divisions.

I Impact of a morphokinetic selection on the outcome of euploid embryos

Introduction The development of incubators with built-in time-lapse technology has enabled continuous non-invasive monitoring of embryo development from fertilization to blastocyst stage and the possibility of appraising the precise timing of embryonic cell divisions. Many studies have tried to connect, more or less conclusively, embryo development, timing of mitotic divisions, embryo viability, aneuploidy and implantation. However, to the best of our knowledge, morphokinetics of euploid embryos have not been explored regarding which time points are critical for their implantation. Material and Methods This observational study was based on 143 patients (mean age: 35.07) applying for preimplantation genetic screening (PGS) due to advanced maternal age (>37) or a history of abnormal fetal karyotype. Patients with structural abnormalities were not included in the analysis. Cases included in the analysis had at least two euploid embryos and all embryos were cultured in a time-lapse incubator (EmbryoScope™). Good or top-quality blastocysts (at least 4BB) were biopsied and analyzed either by next generation sequencing (NGS) (PGM platform, ThermoFisher) or by array Comparative Genomic Hybridization (aCGH, Illumina). Embryo morphokinetics of 57 patients with a negative outcome were compared to 86 patients with an ongoing pregnancy. Results Morphokinetic variables for all cleavage events up to the expanded blastocyst stage were annotated. All relevant events (fertilization, cleavages, morula and blastocyst formation) were checked on a daily basis, and time of cleavage to two-cell embryo (t2) and subsequent divisions t3, t4, t5, t6, t7, t8 and t9+ were recorded in the EmbryoViewer® workstation. The time of all mitotic events was expressed as hours post-ICSI. Error! Not a valid link.The kinetics of euploid embryos was evaluated according to their pregnancy outcome. Implanted euploid embryos were faster for each embryonic event or cleavage time annotated. When median values were compared, a time difference of 0.3-3.28h was found between implanted and non-implanted euploid embryos, statistically significant differences being found for pronuclei appearance (tPNa) (p=0.0065) and the time to achieve 9 cells (t9) (p=0.0132). Conclusions Although many algorithms regarding the deselection of aneuploid embryos have been published, none have focused on the selection of euploid embryos. Our study showed that, overall, implanted euploid embryos were faster for each embryonic event or cleavage time annotated. Additionally, the first event annotated, the pronuclei appearance and last cleavage (t9) are significantly different in implanted and non-implanted euploid embryos. These morphokinetic criteria may be used to prioritize euploid embryos for transfer.

Quantum-like decreased embryogenesis time with increased cold exposure time

Three theoretical models have been proposed to explain lifespan extension resulting from exposure to extreme conditions in microscopic animals: individuals become completely dormant and stop aging, continue to age or age but at a diminished rate. Here we show that the earliest life history stages, embryonic cell divisions, in the tardigrade species Hypsibius dujardini are retarded when eggs are reared at 0 °C. Compared to control specimens reared at 22 °C, juveniles that hatched from eggs exposed to 0 °C for 4 days and returned to 22 °C experienced a three-day lag, indicating that their biological age was less than their chronological age. As cold exposure duration increased (days = 10, 20, 40), incubation period at 22 °C decreased incrementally (days = 3, 2, 1), suggesting that tardigrades involve a threshold-determined, quantum-like, energetic-based system for controlling embryogenesis.

FACT complex gene duplicates exhibit redundant and non-redundant functions in C. elegans

FACT (facilitates chromatin transcription) is a histone chaperone complex important in genomic processes including transcription, DNA replication, and DNA repair. FACT is composed of two proteins, SSRP1 and SPT16, which are highly conserved across eukaryotes. While the mechanisms for FACT in nucleosome reorganization and its relationship to DNA processes is well established, how these roles impact coordination in multicellular animal development are less well understood. Here we characterize the genes encoding FACT complex proteins in the nematode C. elegans. We show that whereas C. elegans includes one SPT16 gene (spt-16), two genes (hmg-3 and hmg-4) encode SSRP1 proteins. Depletion of FACT complex genes interferes with embryonic cell division and cell cycle timing generally, with anterior pharynx development especially sensitive to these defects. hmg-3 and hmg-4 exhibit redundancy for these maternally-provided embryonic functions, but are each uniquely required zygotically for normal germline development. This work provides a framework to study FACT gene function in developmental processes, and identifies that distinct functional requirements for gene duplicates can be manifest within a single tissue.

High sperm DNA fragmentation delays human embryo kinetics when oocytes from young and healthy donors are microinjected.

Time-lapse monitoring (TLM) technology has been implemented in the clinical setting for the culture and selection of human embryos. Many studies have assessed the association between sperm DNA fragmentation (sDNAf) and clinical outcomes after ART, but little is known about the influence of sDNA on embryo morphokinetics. The objective of this retrospective study, which includes 971 embryos from 135 consecutive ICSI cycles (56 cases with own oocytes, 79 with oocytes from young and healthy donors), was to assess if sDNAf has an impact on embryo morphokinetics. Samples used to perform ICSI were analyzed by the flow cytometry TUNEL assay, and embryo development was assessed through an EmbyoScope® system. The association between sDNAf and the timings of cell cleavage was analyzed by categorizing the first variable into quartiles: ≤6.50%; 6.51-10.70%; 10.71-20.15%; >20.15%. In cases where sDNAf was above 20.15% (the upper quartile), embryos derived from donated oocytes (n=644) showed significantly slower divisions. Such association was not observed in embryos obtained from the patients' own oocytes (n=327). The embryo cleavage pattern (either normal, direct from 1 to 3 blastomeres, direct from 1 to 4 blastomeres, incomplete, reversed or asynchronous) was independent of the sDNAf level. Blastocyst arrival rate was 63.0% and the rate of good quality embryos (transferred and frozen embryos divided by the number of zygotes) was 45.49%. Neither parameter was related to the levels of sDNAf. According to our results, the association between high sDNAf and donated oocytes led to delayed cell division. To our knowledge, this is the first study suggesting that sDNAf can delay human embryo cleavage timings when oocytes from donors are inseminated. This finding may indicate that, in the presence of increased DNA damage, time is needed before the first embryonic cell division for the activation of the optimal DNA repairing machinery in higher quality oocytes.

Insight into the mechanisms and consequences of recurrent telomere capture associated with a sub-telomeric deletion.

A complex mosaicism of the short arm of chromosome 1 detected by SNP microarray analysis is described in a patient presenting a 4-Mb 1p36 terminal deletion and associated phenotypic features. The array pattern of chromosome 1p displayed an intriguing increase in divergence of the SNP heterozygote frequency from the expected 50% from the centromere towards the 1p36 breakpoint. This suggests that various overlapping segments of UPD were derived by somatic recombination between the 1p homologues. The most likely explanation was the occurrence of a series of events initiated in either a gamete or an early embryonic cell division involving a 1pter deletion rapidly followed by multiple telomere captures, resulting in additive, stepped increases in frequency of homozygosity towards the telomere. The largest segment involved the entire 1p, and at least four other capture events were observed, indicating that at least five independent telomere captures occurred in separate cell lineages. The determination of breakpoint position by detection of abrupt changes in B-allele frequency using a moving window analysis demonstrated that they were identical in blood and saliva, the tissues available for analysis. We developed a model to explain the interaction of parameters determining the mosaic clones and concluded that, while number, size, and position of telomere captures were important initiating determinants, variation in individual clone frequencies was the main contributor to mosaic differences between tissues. All previous reports of telomere capture have been restricted to single events. Other cases involving multiple telomere capture probably exist but require investigation by SNP microarrays for their detection.

Extremely low incidence of mosaicism in human blastocysts mimics occurrence in natural and IVF clincial pregnancies

Chromosome mis-segregation during embryonic mitotic cell division is responsible for the generation of mosaicism, which is defined as two or more cell populations with different chromosome constitutions within the same embryo. Estimates of embryonic chromosomal mosaicism in the literature have been inconsistent with potentially high rates influenced by clinical procedures, laboratory protocols, and patient variability. The aim of this multicenter study was to evaluate the prevalence of embryonic chromosomal mosaicism at numerous IVF clinics consisting of variable infertility patient demographics. Multi-center prospective study. Initial diagnosis of chromosome numeration from trophectoderm (TE) biopsies was performed using the VeriSeq™ NGS platform (Illumina) at a single genetics laboratory. A total of six IVF clinics situated across North America were included in the multicenter study. Each geographical location is considered to have a unique population demographic and their own patient management for clinical treatment including, ovarian stimulation protocols. Embryos in the six IVF laboratories were cultured under similar optimized conditions utilizing the same consumables, reagents, equipment and comparable biopsy techniques. Statistical analysis included Student’s t-test, Fisher’s Exact and ANOVA where appropriate, significance at P<0.05. The overall incidence of chromosome mosaicism following TE biopsy of 16,352 human blastocysts was observed at only 2.6%. There was no significant variability observed across the six IVF clinics (Clinic A=2.6%; Clinic B=2.4%; Clinic C=2.4%; Clinic D=2.5%; Clinic E=2.1%; Clinic F=2.8%; ns). Advanced reproductive age was not associated with chromosomal mosaicism (mean maternal age = 36.3±4.2years; mean paternal age = 38.2±6.1years). For all infertility patients that were identified to have a mosaic blastocyst in their embryo cohort, the vast majority, two thirds, were younger than 38 years of age (P<0.05). Neither infertility diagnosis, ovarian reserve, blastocyst morphology, day of biopsy, nor the chromosome involved in the error were associated with chromosomal mosaicism. Blastocysts identified with chromosomal mosaicism were not transferred. Multiple re-analysis of mosaic blastocysts confirmed the original mosaicism diagnosis and the identification of the inverse aneuploidy in the re-biopsy sections with random allocation between the TE and inner cell mass. This novel study suggests that chromosomal mosaicism in human blastocysts is indeed extremely low, more closely resembling the incidence of mosaicism in spontaneous and IVF clinical pregnancies (1-2%). The combination of optimized conditions and robust protocols in both the IVF and genetics laboratories contributes to the most successful clinical outcomes for infertility patients, including the expected biological representation of chromosomal mosaicism.

Comparative proteome analysis between C. briggsae embryos and larvae reveals a role of chromatin modification proteins in embryonic cell division

Caenorhabditis briggsae has emerged as a model for comparative biology against model organism C. elegans. Most of its cell fate specifications are completed during embryogenesis whereas its cell growth is achieved mainly in larval stages. The molecular mechanism underlying the drastic developmental changes is poorly understood. To gain insights into the molecular changes between the two stages, we compared the proteomes between the two stages using iTRAQ. We identified a total of 2,791 proteins in the C. briggsae embryos and larvae, 247 of which undergo up- or down-regulation between the two stages. The proteins that are upregulated in the larval stages are enriched in the Gene Ontology categories of energy production, protein translation, and cytoskeleton; whereas those upregulated in the embryonic stage are enriched in the categories of chromatin dynamics and posttranslational modification, suggesting a more active chromatin modification in the embryos than in the larva. Perturbation of a subset of chromatin modifiers followed by cell lineage analysis suggests their roles in controlling cell division pace. Taken together, we demonstrate a general molecular switch from chromatin modification to metabolism during the transition from C. briggsae embryonic to its larval stages using iTRAQ approach. The switch might be conserved across metazoans.

Night Monkey Hybrids Exhibit De Novo Genomic and Karyotypic Alterations: The First Such Case in Primates.

Using molecular chromosomal analyses, we discovered night monkey hybrids produced in captivity from matings between a female Aotus azarae boliviensis (2n = 50) and a male Aotus lemurinus griseimembra (2n = 53). The parents produced seven offspring in total, including one male and six females—a pattern consistent with Haldane’s rule. Chromosomal studies were conducted on four of the hybrid offspring. Two of them showed relatively “simple” mixture karyotypes, including different chromosome numbers (2n = 51, 52), which were formed because of a heteromorphic autosome pair in the father (n = 26, 27). The other two hybrid monkeys exhibited de novo genomic and karyotypic alterations. Detailed analysis of the alterations revealed that one individual carried a mixture karyotype of the two parental species and an X chromosome trisomy (53,XXX). The second individual displayed trisomy of chromosome 18 (52,XX,+18) and a reciprocal translocation between autosomes 21 and 23 (52,XX,+18,t(21;23)). Interestingly, the second monkey exhibited mosaicism among blood cells (mos52,XX,+18[87]/52,XX,+18,t(21;23)[85]), but only a single karyotype (52,XX,+18) in skin fibroblast cells. The X- and 18-trisomies were derived from a doubling of the mother’s chromosomes in early embryonic cell division, and the reciprocal translocation likely developed in the bone marrow of the offspring, considering that it was observed only in blood cells. Such occurrence of trisomies in hybrid individuals is a unique finding in placental mammals.

Spatiotemporal coupling and decoupling of gene transcription with DNA replication origins during embryogenesis in C. elegans.

The primary task of developing embryos is genome replication, yet how DNA replication is integrated with the profound cellular changes that occur through development is largely unknown. Using an approach to map DNA replication at high resolution in C. elegans, we show that replication origins are marked with specific histone modifications that define gene enhancers. We demonstrate that the level of enhancer associated modifications scale with the efficiency at which the origin is utilized. By mapping replication origins at different developmental stages, we show that the positions and activity of origins is largely invariant through embryogenesis. Contrary to expectation, we find that replication origins are specified prior to the broad onset of zygotic transcription, yet when transcription initiates it does so in close proximity to the pre-defined replication origins. Transcription and DNA replication origins are correlated, but the association breaks down when embryonic cell division ceases. Collectively, our data indicate that replication origins are fundamental organizers and regulators of gene activity through embryonic development.

Systems biology: Molecular memoirs of a cellular family.

A system that introduces random modifications to barcode sequences embedded in cells' DNA allows lineage relationships between cells to be discerned, while preserving the cells' spatial relationships. See Letter p.107 It is difficult to establish cell division history and lineage relationships for tissues that are not easily accessed. DNA barcoding approaches can provide this information, but do not offer spatial data. This collaboration between the labs of Long Cai and Michael Elowitz harnesses the power of CRISPR/Cas9-mediated mutagenesis and multiplexed single-molecule RNA fluorescence hybridization (smFISH) to build a new tool, termed MEMOIR, which they use to follow mouse embryonic stem cell divisions. MEMOIR gives access to lineage information in situ and at the single-cell level, while at the same time monitoring changes in gene expression state.

Chondroitin 4-O-Sulfotransferase Is Indispensable for Sulfation of Chondroitin and Plays an Important Role in Maintaining Normal Life Span and Oxidative Stress Responses in Nematodes

Chondroitin sulfate (CS)/chondroitin (Chn) chains are indispensable for embryonic cell division and cytokinesis in the early developmental stages in Caenorhabditis elegans and mice, whereas heparan sulfate (HS) is essential for axon guidance during nervous system development. These data indicate that the fundamental functions of CS and HS are conserved from worms to mammals and that the function of CS/Chn differs from that of HS. Although previous studies have shown that C. elegans produces HS and non-sulfated Chn, whether the organism produces CS remains unclear. Here, we demonstrate that C. elegans produces a small amount of 4-O-sulfated Chn and report the identification of C41C4.1, an orthologue of the human chondroitin 4-O-sulfotransferase gene. Loss of C41C4.1 in C. elegans resulted in a decline in 4-O-sulfation of CS and an increase in the number of sulfated units in HS. C41C4.1 deletion mutants exhibited reduced survival rates after synchronization with sodium hypochlorite. Collectively, these results show for the first time that CS glycans are present in C. elegans and that the Chn 4-O-sulfotransferase responsible for the sulfation plays an important role in protecting nematodes from oxidative stress.

Caenorhabditis elegans glutamylating enzymes function redundantly in male mating.

ABSTRACTMicrotubule glutamylation is an important modulator of microtubule function and has been implicated in the regulation of centriole stability, neuronal outgrowth and cilia motility. Glutamylation of the microtubules is catalyzed by a family of tubulin tyrosine ligase-like (TTLL) enzymes. Analysis of individual TTLL enzymes has led to an understanding of their specific functions, but how activities of the TTLL enzymes are coordinated to spatially and temporally regulate glutamylation remains relatively unexplored. We have undertaken an analysis of the glutamylating TTLL enzymes in C. elegans. We find that although all five TTLL enzymes are expressed in the embryo and adult worm, loss of individual enzymes does not perturb microtubule function in embryonic cell divisions. Moreover, normal dye-filling, osmotic avoidance and male mating behavior indicate the presence of functional amphid cilia and male-specific neurons. A ttll-4(tm3310); ttll-11(tm4059); ttll-5(tm3360) triple mutant, however, shows reduced male mating efficiency due to a defect in the response step, suggesting that these three enzymes function redundantly, and that glutamylation is required for proper function of the male-specific neurons.

The subcortical maternal complex: multiple functions for one biological structure?

The subcortical maternal complex (SCMC) is a multiprotein complex uniquely expressed in mammalian oocytes and early embryos, essential for zygote progression beyond the first embryonic cell divisions. Similiar to other factors encoded by maternal effect genes, the physiological role of SCMC remains unclear, although recent evidence has provided important molecular insights into different possible functions. Its potential involvement in human fertility is attracting increasing attention; however, the complete story is far from being told. The present mini review provides an overview of recent findings related to the SCMC and discusses its potential physiological role/s with the aim of inspiring new directions for future research.

Raman-based noninvasive metabolic profile evaluation ofin vitrobovine embryos

The timing of the first embryonic cell divisions may predict the ability of an embryo to establish pregnancy. Similarly, metabolic profiles may be markers of embryonic viability. However, in bovine, data about the metabolomics profile of these embryos are still not available. In the present work, we describe Raman-based metabolomic profiles of culture media of bovine embryos with different developmental kinetics (fast x slow) throughout the in vitro culture. The principal component analysis enabled us to classify embryos with different developmental kinetics since they presented specific spectroscopic profiles for each evaluated time point. We noticed that bands at 1076 cm(−1) (lipids), 1300 cm(−1) (Amide III), and 2719 cm(−1) (DNA nitrogen bases) gave the most relevant spectral features, enabling the separation between fast and slow groups. Bands at 1001 cm(−1) (phenylalanine) and 2892 cm(−1) (methylene group of the polymethylene chain) presented specific patterns related to embryonic stage and can be considered as biomarkers of embryonic development by Raman spectroscopy. The culture media analysis by Raman spectroscopy proved to be a simple and sensitive technique that can be applied with high efficiency to characterize the profiles of in vitro produced bovine embryos with different development kinetics and different stages of development.

Expression profiles of long and short rnas in the cytoplasm and nuclei of growing chicken (Gallus gallus domesticus) oocytes

Maternal RNAs accumulated during oocyte maturation are required not only for zygote formation but also for supporting the first embryonic cell divisions until the activation of the embryo genome. The key stages of transcriptome analysis include the adaptation of RNA extraction procedures and characterization of the RNA expression profile. The ovaries of domestic birds represent an adequate model for the exploration of RNA accumulation during oogenesis. In the present study, we optimized the methodologies for RNA extraction from the oocyte cytoplasm and nucleoplasm of egg-laying chickens (Gallus gallus domesticus), and characterized the changes in the expression profiles of long and short RNAs during oocyte growth. The cytoplasmic RNA fractions contained 28S and 18S ribosomal RNAs (rRNAs), small RNAs, and long RNAs that were heterogeneous in size. The profiles of the total RNAs from growing oocyte nuclei were dominated by low molecular weight RNAs corresponding in size to transport RNAs, small nuclear RNAs, and short regulatory RNAs. Importantly, nuclei from chicken growing oocytes demonstrated trace amounts or the absence of 28S and 18S rRNAs, which was possibly due to the inactivation of the single nucleolus organizer. We identified three groups of short RNAs differing in size (from 20 to 40 nucleotides) in the chicken oocytes. These may correspond to short regulatory RNA classes: microRNA, siRNA and piRNA. Furthermore, we demonstrated that short RNAs accumulate in the cytoplasm during oocyte growth. We propose that the short RNAs accumulated in the avian oocyte cytoplasm are involved in the regulation of genome functions during the early embryogenesis stages.

CSR-1 Slices a Balance

CSR-1 is a germline-expressed C. elegans Argonaute protein essential for viability. In this issue of Cell, Gerson-Gurwitz et al. now demonstrate a role for CSR-1 and its slicer activity in downregulating the levels of maternally deposited mRNAs to fine-tune the expression of proteins with critical roles in embryonic cell divisions.

Molecular Reproduction &amp; Development Volume 83, Issue 4, April 2016

Embryo morphokinetics suggest that the timing of the first embryonic cell divisions may predict the developmental potential of an embryo. Milazzotto et al (this issue) identified the transcriptomic and metabolic differences between bovine embryos with different morphokinetics, suggesting that sorting embryos based on cleavage speed may select for different metabolic patterns.

Early cleavages influence the molecular and the metabolic pattern of individually cultured bovine blastocysts.

Embryo morphokinetics suggests that the timing of the first embryonic cell divisions may predict the developmental potential of an embryo; however, correlations between embryonic morphokinetics and physiology are not clear. Here, we used RNA sequencing to determine the gene expression profile of in vitro-produced early- and late-dividing bovine embryos and their respective blastocysts, and compared these profiles to in vivo-produced blastocysts to identify differentially expressed genes (DEGs). Principal component analysis revealed that fast- and slow-dividing embryos possess similar transcript abundance over the first cleavages. By the blastocyst stage, however, more DEGs were observed between the fast- and slow-dividing embryo groups, whereas blastocysts from the slow-dividing group were more similar to in vivo-produced blastocysts. Gene ontology enrichment analysis showed that the slow-dividing and in vivo-produced blastocysts shared biological processes related to groups of up- or down-regulated genes when compared to the fast-dividing blastocysts. Based on these DEG results, we characterized the relationship between developmental kinetics and energy metabolism of in vitro-produced bovine embryos. Embryos from fast- and slow-dividing groups exhibited different pyruvate and lactate metabolism at 22 hr post-in vitro culture (hpc), glucose consumption at 96 hpc, and glutamate metabolism at 168 hpc. Glycogen storage was similar between cleavage-stage and morulae groups, but was higher in the blastocysts of the slow-dividing group. On the other hand, blastocysts of the fast-dividing group had a higher concentration of lipids. Taken together, these data identify transcriptomic and metabolic differences between embryos with different morphokinetics, suggesting that sorting embryos based on cleavage speed may select for different metabolic patterns. Mol. Reprod. Dev. 83: 324-336, 2016. © 2016 Wiley Periodicals, Inc.