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Saitou, M.

Publications and source records attributed to Saitou, M..

At least 19 recordsLinked to original sources

CycloCross: Adversarial single-cell RNA-seq data translation across species

Single-cell RNA sequencing (scRNA-seq) has transformed our understanding of cellular heterogeneity, yet most datasets are limited to a handful of model organisms, leaving critical gaps in cross-species biology. Existing computational methods for integrating multi-species scRNA-seq data often rely on one-to-one ortholog mapping, which fails to account for gene duplications, losses, or functional divergences. To address these challenges, we introduce CycloCross, an adversarial method based on the CycleGAN architecture, designed to translate scRNA-seq data between species without requiring a priori gene correspondences. Unlike generative models that synthesize data from noise, CycloCross leverages real data from a source species, enabling more realistic and data-efficient translations. We demonstrate its effectiveness in germ cell development, translating data between mouse, macaque, and human with conserved yet divergent transcriptional programs. CycloCross not only preserves temporal dynamics but also enables interpolation and extrapolation of gene expression at missing time points, offering predictions for experimentally inaccessible states. Furthermore, CycloCross generates biologically plausible samples even with limited data, outperforming traditional generative models.

bioinformatics↗

Imputed graph-genotyped structural variants identify regulatory haplotypes associated with gene expression in Atlantic salmon

Structural variants (SVs) can affect gene regulation, but they are difficult to include in expression genetic studies when large RNA-seq cohorts lack whole-genome sequencing. This is common in non-human and non-model systems, where whole-genome sequencing at population scale remains costly. As a result, expression quantitative trait locus (eQTL) studies often rely on single nucleotide polymorphism (SNP) markers. These analyses can identify expression-associated regions, but often provide limited biological interpretation of the underlying regulatory mechanisms. Here, we used Atlantic salmon as a study system to test whether graph-genotyped SVs can be imputed into a SNP-array-genotyped RNA-seq cohort and used to interpret regulatory haplotypes. SVs were discovered from two long-read-sequenced individuals, supplemented with short-read SV and SNP calls from a 112-individual whole-genome-sequenced reference panel, graph-genotyped, jointly phased with SNPs, and imputed into 906 offspring with gill RNA-seq and SNP-array genotypes. After size filtering, the imputed SV catalogue contained 100,269 variants and showed nonuniform genomic distributions associated with sex-specific recombination landscapes. Association testing identified 51 SV-eQTL candidates, including 35 cis and 16 trans associations. These candidates were enriched for short-read-derived variants, indicating that short-read supplementation can recover regulatory variants missed by small-scale long-read discovery. SV-eQTL candidates were more strongly tagged by nearby SNPs than non-associated variants generally, but individual SNP lead markers often failed to capture the same eQTL signals in conditional regression. Retained candidates after the conditional analysis included target-gene-overlapping deletions, nearby local variants without target-gene overlap, trans associations, and short insertions with opposite effects on gene expression. These results show that imputed graph-genotyped SVs can add biological interpretation to possible regulatory haplotypes.

genomics↗

Ancient persistence and recurrent emergence of structural variants across divergent Atlantic salmon lineages

Structural variants (SVs) are a major source of genomic diversity, yet the evolutionary origins of SVs shared across divergent populations remain difficult to resolve. Shared SVs may reflect ancient polymorphism, recurrent mutation, introgression, or subsequent lineage-specific frequency change, but the relative contribution of these processes often remains difficult to distinguish. Here, we investigated SV evolution across four Atlantic salmon (Salmo salar) lineages differing in geography, Europe versus North America, and domestication status, wild versus farmed. Using sensitive SV discovery, stringent genotyping, local PCA, haplotype-distance analyses, and forward simulations, we tested whether broadly shared SVs behave as a single class of variation or separate into distinct evolutionary categories. We generated a high-confidence SV map and found that SVs were enriched in repetitive regions, particularly segmental duplications and LTR retrotransposons, consistent with genome architecture shaping SV formation. Nearly half of high-confidence SVs were shared across all four lineages despite deep continental divergence, and simulations showed that this broad sharing is more consistent with ancient persistence than recurrent mutation alone. In contrast, a small subset of large SVs exhibited complex PCA clustering and multimodal haplotype-distance distributions, consistent with recurrent formation at structurally unstable loci. Large SVs also showed contrasting frequency trajectories between continents, and one immune gene-rich copy-number variable region showed a marked frequency increase in domesticated European salmon. Together, these results show that shared SVs comprise distinct evolutionary categories shaped by ancient persistence, recurrent mutation, and lineage-specific frequency change.

genomics↗

No-take zone effectiveness in linear coastal systems depends on spatially heterogeneous placement and larval dispersal directionality

Marine protected areas (MPAs) are widely implemented to protect and/or rebuild exploited populations, yet their population-level effectiveness remains highly variable. Key unresolved knowledge gaps include how the size and the spatial placement of no-take zones interacts with larval dispersal directionality, particularly in linear coastal systems where connectivity is asymmetric. For sedentary species with planktonic larvae, such as European lobster, it is unclear under which dispersal regimes spatial configuration of protection critically determines positive demographic outcomes. Here, we address this gap using a spatially explicit individual-based model parameterized for the European lobster. We ask (i) whether no-take zones consistently enhance abundance and size structure relative to fished areas, and (ii) whether the positioning of no-take and open areas affects spatial protection while holding total protected area constant and (iii) how the alignment between larval dispersal direction and the positioning of no-take areas influences protection outcome. We contrast local, symmetric long-distance, and strongly unidirectional larval dispersal across alternative MPA layouts with equal total protected area but with different spacing. We show that no-take zones reliably increase abundance and the prevalence of large individuals. However, when larval dispersal is strongly unidirectional, population recovery depends on reserve placement: downstream no-take zones benefit from both larval import and local retention, whereas upstream reserves primarily export reproductive output and show limited local recovery. These results indicate that reserve performance cannot be evaluated independently of connectivity structure and identify dispersal directionality as a key determinant of when and where spatial configuration matters for MPA effectiveness in linear coastal systems.

ecology↗

Resampling-based validation of a SNP panel for hybrid detection across generations: a case study in European lobster

Accurate detection of hybridization and introgression is critical for both evolutionary research and applied conservation. In many systems, however, hybrid ancestry is difficult to detect beyond the F1 generation, especially when based on limited genetic markers. In European waters, hybridization between the native Homarus gammarus and the invasive H. americanus poses a direct risk to the integrity of native stocks and effective fishery management, yet detection methods are often limited to morphological traits or first-generation hybrids. A set of 79 SNPs previously developed to distinguish species between American and European lobsters and F1 individuals has shown promise, but its capacity to resolve later-generation backcrosses remains untested. Here, we present a resampling-based evaluation of this panels performance under realistic introgression scenarios, using individual-based population genetic models informed by empirical data. We show that the panel retains discriminatory power across multiple hybrid classes, with diminishing accuracy in second-generation backcrosses. These findings validate the panels utility for applied monitoring and highlight the broader potential of resumpling-anchored frameworks to benchmark hybrid detection tools in a wide range of species. Article summaryThis study tests how well a reduced panel of genetic markers can detect hybridization across multiple generations. Using empirical genetic data of a 79-SNP panel from European and American lobsters, the authors generated individuals with known ancestry proportions through a resampling framework that preserves observed genetic variation. These data were analysed using model-based genetic assignment and ordination. The results show that the marker panel reliably identifies pure species and first-generation hybrids, but has reduced power to distinguish later backcross generations, mainly between adjacent hybrid classes. The study provides a practical benchmark for evaluating reduced marker panels used in applied monitoring and conservation genetics.

genomics↗

Comparative transcriptomic analysis defines shared and mammary-specific gene expression programs across glandular tissues

The mammary gland represents a defining evolutionary innovation of mammals, which is an epithelial tissue with specialized secretory functions that are regulated by hormonal and transcriptional processes. While mammary differentiation has been extensively studied, its transcriptional relationship to other epithelial tissues with secretory activity has been less explored in an evolutionary context. Here, we used bulk RNA-seq data from the GTEx project to examine gene expression patterns in human breast tissue alongside a set of other tissues that contain epithelial secretory components, as well as non-glandular tissues. Using cross-tissue differential expression analyses, co-expression network analysis, and integration with single-cell expression references, we characterized patterns of gene expression that recur across multiple secretory tissues and those that show relative enrichment in breast tissue. This analysis highlights three broad classes of expression patterns: genes with elevated expression across several secretory tissues, genes showing preferential expression in breast tissue, and genes displaying sex-associated differences in breast samples. These gene sets are associated with distinct functional annotations, including epithelial structure, metabolic and hormone-responsive processes, and immune-related functions. Comparative analyses of gene conservation indicate that many genes preferentially expressed in breast tissue are evolutionarily conserved across vertebrates, consistent with the reuse of pre-existing gene repertoires in the mammalian lineage in tissue-specific contexts. Together, these results support a model in which breast tissue identity reflects the regulatory re-weighting and integration of conserved epithelial secretory modules rather than lineage-specific gene innovation. By placing human breast tissue gene expression in a comparative and evolutionary context, this study provides a molecular framework for understanding how complex organ-level traits arise through regulatory modularity. SignificanceThe mammary gland is a defining feature of mammals, yet it remains unclear how its gene activity relates to that of other tissues with similar secretory functions. By comparing gene expression in human breast tissue with other epithelial secretory tissues, this study shows that much of breast tissue gene expression is shared with a common epithelial program, while a smaller set of genes is associated with breast-specific metabolic and hormonal functions. These findings help clarify how mammary gland specialization arises in mammalian lineages from ancestral genetic materials rather than from entirely new gene emergences, providing a framework for understanding the evolution of new organs.

genomics↗

Large-scale eQTL analyses in Atlantic salmon reveal persistent dosage compensation 100 million years after genome duplication.

Whole-genome duplication (WGD) through autopolyploidization has played a role in genome evolution across eukaryotes. A major consequence of WGD is the rewiring of gene regulatory networks, partly driven by selection on dosage balance. In multicellular organisms, evidence for dosage balance selection has relied on comparative patterns of duplicate gene retention and expression, with few studies directly examining regulatory architecture after WGD. Here, we analysed a large-scale eQTL dataset from Atlantic salmon (Salmo salar), which experienced a WGD 100 million years ago. We found that trans-regulatory connections were enriched between duplicated regions, indicating long-term conservation of ancestral interchromosomal regulatory interactions. Overall, 230 duplicated genes (5%) shared eQTLs, suggesting conserved regulatory control. Moreover, 16 gene pairs showed compensatory expression effects mediated by a common regulator, consistent with predictions of the dosage balance hypothesis. These gene pairs were significantly enriched in recently rediploidized regions. Our results indicate long-term maintenance of dosage balance after WGD. TeaserGenetic regulation in Atlantic salmon shows that duplicated genes can remain dosage-balanced across 100 million years of evolution.

evolutionary biology↗

Geometric inference of cross-species transcriptome correspondence using Gromov-Wasserstein optimal transport

Sequence homology reveals evolutionary gene correspondences, but identifying functionally corresponding genes within specific cell types or during cell fate specification remains non-trivial. Here, frameworks that translate insights from model organisms into other species would accelerate wet-lab studies, so we develop Species-OT, a cross-species transcriptome analysis framework using Gromov-Wasserstein optimal transport, which quantitatively compares the geometry of transcriptome distributions. Given a pair of bulk or single-cell RNA-sequencing datasets, Species-OT returns a gene-to-gene correspondence capturing probabilistic alignments of regulatory roles, and a transcriptomic distance quantifying overall divergence. Applied pairwise, Species-OT yields a transcriptomic discrepancy array and a hierarchical clustering tree analogous to a phylogenetic tree. We validate Species-OT using bulk RNA-seq data for cell fate specification from pluripotent stem cells in mice, monkeys, and humans and scRNA-seq data from pluripotent stem cells of six mammalian species. Species-OT identifies gene correspondences within cell fate specification, while transcriptomic discrepancies recapitulate expected species relationships.

bioinformatics↗

A major locus on chromosome 14 impacts developmental variation of Atlantic salmon smoltification

BackgroundSmoltification in anadromous Atlantic salmon is a complex developmental process involving physiological and cellular changes that enable freshwater fish to adapt to seawater. Central to this transformation is the functional transformation of the gill to manage osmoregulatory demands. While environmental cues like photoperiod are known to influence smolt development, the genetic architecture underlying smolt development -- particularly related to gill physiology -- remains poorly understood. ResultsA large-scale eQTL analysis across 3,000 Atlantic salmon subjected to three photoperiod regimes, identified over 45,000 significant SNP-gene expression associations in gill tissue. Notably, we discovered a 0.5 Mbp large trans-eQTL hotspot on chromosome 14. This hub-locus was associated with expression of more than 2,000 genes across the genome which were significantly enriched for gill cell type specific markers. In addition we found that the hub-locus was associated with somatic growth. Our findings support a "local tissue effect" model, where cis-regulatory or protein sequence variants within the hub-locus modulate cell proliferation and differentiation of gill cell types. ConclusionThis work advances our understanding of the genetic basis of smoltification in Atlantic salmon and provides a foundation for future studies using single-cell approaches to resolve cell-type specific mechanisms underlying genetic variation in smolt development.

genetics↗

Gene gain and loss drive the diversification of gig immune genes in teleosts: structural and regulatory insights from Atlantic salmon

Interferon-stimulated genes (ISGs) are key players in vertebrate antiviral immunity. Among teleost ISGs, the grass carp reovirus-induced gene (gig) families 1 and 2 (gig1 and gig2, respectively) are absent in mammals but conserved in fishes and amphibians, and they have been implicated in resistance to viral infections across several aquaculture species. In particular, gig1 and gig2 genes are transcriptionally induced by viral stimuli in teleosts such as zebrafish, grass carp, and salmonids, and recent studies have highlighted their potential involvement in resistance to economically important diseases like pancreas disease in Atlantic salmon. Yet, the rapid evolution of these genes hinders a comprehensive understanding of their diversification process and regulatory mechanisms. This study investigated gig gene evolution across teleosts, with a focus on Atlantic salmon (Salmo salar). Phylogenetic analysis across representative ray-finned fishes (Actinopterygii), including both teleosts and non-teleost outgroups such as the spotted gar (Holostei), indicated that gig1 is restricted to teleosts, with no identifiable homologs in non-teleost lineages. In contrast, gig2 genes are present in both teleosts and the spotted gar, suggesting an origin prior to the teleost-specific whole genome duplication (Ts3R), likely in early non-amniote vertebrates. Whole-genome duplication drove lineage-specific expansions, particularly of gig2 in salmonids. Structural and transcriptomic analyses showed that gig1 and gig2 differ in domain composition, repeat content, and regulation. Our findings suggest the complex interplay of duplication history, structural divergence, and transcriptional regulation in shaping immune gene repertoires in teleosts, with implications for understanding host-pathogen interactions and aquaculture disease responses. Article summaryThis study investigates the evolutionary history and diversification of the gig immune gene families in aquatic species, with particular focus on Atlantic salmon. Phylogenetic and structural analyses revealed that gig1 and gig2 follow distinct evolutionary trajectories, shaped by whole-genome and tandem duplications. Further analysis of Atlantic salmon gig genes showed divergent structures and regulation, highlighting a general role of gig genes in antiviral Interferon-mediated immunity and additionally suggesting functional specialization across gig paralogs. Together, these findings improve our understanding of immune gene evolution in fishes and provide insights relevant to antiviral defense and disease management in aquaculture species.

genomics↗

Impaired trap closure in the counting-deficient Venus flytrap mutant DYSCALCULIA is caused by cell wall biomechanics

Living in nutrient-poor environments, the carnivorous Venus flytrap Dionaea muscipula captures animal prey to compensate for this deficiency. Stimulation of trigger hairs located on the inner trap surface elicits an action potential (AP). While two consecutive APs result in fast trap closure in wildtype (WT) plants, sustained AP generation by the insect struggling to escape the trap leads to jasmonic acid (JA) biosynthesis, formation of the digestive "stomach", and release of enzymes needed to decompose the victim. The Dionaea muscipula DYSCALCULIA (DYSC) mutant is able to fire touch-induced APs, but unlike WT plants, it does not snap-close its traps after two consecutive APs. Moreover, DYSC plants fail to properly initiate the JA pathway in response to mechanostimulation and even wounding, a well-known JA-dependent process conserved among plants. As demonstrated in previous studies, this DYSC mutant defect is associated with impaired decoding of mechanostimulation (i.e. touch) -induced Ca2+ signals. External JA application to the trap, however, restores slow trap closure and digestive gland function in DYSC, while rapid trap closure is JA-independent and cannot be rescued by exogenous JA application. Higher frequency mechanostimulation and thus more APs, however, revealed that DYSC is still able to close its traps, albeit much slower than WT plants. To reveal the molecular underpinnings of DYSCs delayed trap movement, we generated a chromosome-scale Dionaea genome assembly and profiled gene expression. The refined transcriptomic analysis uncovered widespread misregulation of cell wall-related genes in DYSC, implicating altered cell wall plasticity in the sluggish mutant. Cell indentation studies by atomic force microscopy revealed a strictly localized and strikingly enhanced stiffening of the cell wall for DYSC that may hinder rapid trap closure and snap buckling. Together, these genomic, transcriptomic, and biophysical data identify cell wall elasticity as a key constraint on voltage and Ca2+ dependent trap kinetics. This finding documents the interrelationship between mechanosensing and Ca2+ signaling in the ultrafast capture organ of the Venus flytrap.

plant biology↗

Metatranscriptomic Insights into Microbial Dynamics Prior to Disease Onset in Atlantic Salmon Aquaculture

Poly(A)-selected RNA-seq datasets are routinely generated in aquaculture research, yet the microbial information contained in unmapped reads is seldom explored due to the low abundance of nonhost transcripts and concerns about contamination. In this study, we repurposed Atlantic salmon gill RNA-seq data to assess whether meaningful microbial signals can be recovered using a contamination-aware and compositionally appropriate framework. Unmapped reads were analyzed with a custom Kraken2 database composed exclusively of complete, circularized salmon-associated bacterial genomes together with all available Atlantic salmon assemblies and the human genome. Although microbial sequences represented only a small fraction of total reads, 21 genera were detectable across samples. Genus-level profiles, Jaccard-based ordination, and ANCOM-BC analyses consistently revealed clear differences between tanks, whereas no associations were observed for sex or survival status. Three species exhibited significant tank-specific effects, indicating that environmental factors contributed the strongest detectable structure in the data. The limited microbial diversity recovered here reflects the expected constraints of poly(A)-enriched libraries, yet the results demonstrate that unmapped reads from host-derived RNA-seq can still provide informative environmental signatures when analyzed with curated reference databases and compositional statistical approaches. This strategy offers a practical means to extract exploratory microbiome information from existing transcriptomic datasets.

microbiology↗

Switch-like Gene Expression Modulates Disease Susceptibility

A fundamental challenge in biomedicine is understanding the mechanisms predisposing individuals to disease. While previous research has suggested that switch-like gene expression is crucial in driving biological variation and disease susceptibility, a systematic analysis across multiple tissues is still lacking. By analyzing transcriptomes from 943 individuals across 27 tissues, we identified 1,013 switch-like genes. We found that only 31 (3.1%) of these genes exhibit switch-like behavior across all tissues. These universally switch-like genes appear to be genetically driven, with large exonic genomic structural variants explaining five ([~]18%) of them. The remaining switch-like genes exhibit tissue-specific expression patterns. Notably, tissue-specific switch-like genes tend to be switched on or off in unison within individuals, likely under the influence of tissue-specific master regulators, including hormonal signals. Among our most significant findings, we identified hundreds of concordantly switched-off genes in the stomach and vagina that are linked to gastric cancer (41-fold, p<10-4) and vaginal atrophy (44-fold, p<10-4), respectively. Experimental analysis of vaginal tissues revealed that low systemic levels of estrogen lead to a significant reduction in both the epithelial thickness and the expression of the switch-like gene ALOX12. We propose a model wherein the switching off of driver genes in basal and parabasal epithelium suppresses cell proliferation therein, leading to epithelial thinning and, therefore, vaginal atrophy. Our findings underscore the significant biomedical implications of switch-like gene expression and lay the groundwork for potential diagnostic and therapeutic applications.

bioinformatics↗

Parallel selection in domesticated Atlantic salmon from divergent founders including parallel selection on WGD-derived homeologous regions

Aquaculture has a considerably shorter history compared to the domestication of plants and animals. Among aquatic species, those that have undergone whole genome duplication events (WGD) seem particularly successful. This suggests that genetic redundancy from WGD is important for domestication, possibly similar to plant domestication. Atlantic salmon (Salmo salar), which has experienced a lineage-specific WGD, has undergone rapid domestication through intensive breeding since the 1960s. Here, we examined the genomic responses to the domestication of Atlantic salmon, including the impacts of WGD, by comparing the whole genome sequence data of aquaculture and wild populations from two lineages: the Eastern and Western Atlantic (Western Norway and North America). Our analysis revealed shared selective sweeps on identical SNPs in major histocompatibility complex (MHC) genes across distinct aquaculture populations compared to their wild counterparts. This SNP level parallelism suggests that a combination of long-term balancing selection and recent human-induced selection has significantly shaped the evolutionary trajectory of MHC genes. In addition, we observed selective sweeps on gene pairs in the homeologous regions originating from WGD, highlighting WGDs role in maintaining genomic variation and potentially reducing pleiotropy through sub-functionalization. This unique type of "parallel" selection contributes to adapting to the intensive artificial conditions of aquaculture. These findings provide valuable insights into the genetic mechanisms of domestication and adaptive responses in Atlantic salmon, suggesting that the salmonid whole genome duplication has underpinned their successful rapid domestication. Our research emphasizes the importance of maintaining genetic diversity to support sustainable aquaculture practices.

genomics↗

Atlas of Cynomolgus Macaque Hematopoiesis

Self-renewal and differentiation are inherent properties of hematopoietic stem cells (HSCs) that are necessary to support hematopoiesis; however, the underlying mechanisms, especially in human, remain unclear. Here, using the cynomolgus macaque as a surrogate model, we develop a new gating strategy to isolate with high purity transplantable cynomolgus HSCs and generated a single-cell transcriptomic map of cynomolgus HSCs and progenitor cells -- that covering gestational periods previously not analyzed in human. We show that hematopoietic cells from the late-1st to early-3rd trimester fetal liver and late-2nd trimester and thereafter bone marrow have repopulating potential, closely mimicking humans. Unexpectedly however, we found unlike in human, cynomolgus HSCs express CD38 but not CD33, indicating that these cellular counterparts are molecularly distinct. Our transcriptomic analysis reveals the presence of a direct differentiation pathway from HSCs to megakaryocyte lineages, lineage-primed multipotent progenitors and also identified putative HSC surface markers. Taken together, our comprehensive dataset highlights not only the utility of cynomolgus monkeys as model systems to study hematopoiesis but also their potential for translational applications.

cell biology↗

scEGOT: Single-cell trajectory inference framework based on entropic Gaussian mixture optimal transport

Time-series scRNA-seq data have opened a door to elucidate cell differentiation, and in this context, the optimal transport theory has been attracting much attention. However, there remain critical issues in interpretability and computational cost. We present scEGOT, a comprehensive framework for single-cell trajectory inference, as a generative model with high interpretability and low computational cost. Applied to the human primordial germ cell-like cell (PGCLC) induction system, scEGOT identified the PGCLC progenitor population and bifurcation time of segregation. Our analysis shows TFAP2A is insufficient for identifying PGCLC progenitors, requiring NKX1-2. Additionally, MESP1 and GATA6 are also crucial for PGCLC/somatic cell segregation. These findings shed light on the mechanism that segregates PGCLC from somatic lineages. Notably, not limited to scRNA-seq, scEGOTs versatility can extend to general single-cell data like scATAC-seq, and hence has the potential to revolutionize our understanding of such datasets and, thereby also, developmental biology.

bioinformatics↗

Functional and regulatory diversification of circadian rhythm period genes during the evolution of vertebrates

The Period genes (Per) play essential roles in modulating the molecular circadian clock timing in a broad range of species, which regulates the physiological and cellular through the transcription-translation feedback loop. While the Period gene paralogs are widely observed among vertebrates, the evolutionary history and the functional diversification of Per genes across vertebrates are not well known. In this study, we comprehensively investigated the evolution of Per genes, including de novo binding motif discovery by comparative genomics. We also determined the lineage-specific transcriptome landscape across tissues and developmental stages and phenotypic effects in public RNA-seq data sets of model species. We observed multiple lineage-specific gain and loss events of Per genes, though no simple association was observed between ecological factors and Per gene numbers in each species. Among salmonid fish species, the per3 gene has been lost in the majority, whereas those retaining the per3 gene exhibit not a signature of relaxed selective constraint but rather a signature of intensified selection. We also determined the signature of adaptive diversification of the CRY-binding region in Per1 and Per3, which modulates the circadian rhythm. We also discovered putative regulatory sequences, which are lineage-specific, suggesting that these cis-regulatory elements may have evolved rapidly and divergently across different lineages. Collectively, our findings revealed the evolution of Per genes and their fine-tuned contribution to the plastic and precise regulation of circadian rhythms in various vertebrate taxa. SignificanceThe Period (Per) genes play essential roles in the circadian rhythm in animals. In this study, we comprehensively investigated the evolutionary diversification of the three types of Period genes in vertebrates. As a result, we observed a rapid evolution and sub-functionalization of these genes, especially adaptive diversification signatures in the protein-binding region, which plays a crucial role in regulating circadian rhythms. This underscores the fine-tuned contribution of Per genes in the biological clocks precision and adaptability across various vertebrate taxa.

evolutionary biology↗

Comprehensive comparison of female germ cell development in vitro and in vivo identifies epigenetic gene regulation crucial for oocyte development and embryonic competence

Germ cells are the origin of new individuals. Hence, specifying germ cell identity is crucial for reproduction. The recent establishment of in vitro culture systems for generating oocytes from mouse pluripotent stem cells provides a basis for progress in studies of oogenesis and reproductive technology. However, currently the developmental competence of in vitro generated oocytes is low compared to in vivo grown oocytes. The causes underlying poor oocyte quality remain to be determined. By reconstituting germ cell development in culture from different developmental starting points within gametogenesis, we show that the differentiation of primordial germ cells (PGCs) and primordial germ cell-like cells (PGCLCs) to growing oocytes (GROs), as well as the subsequent growth of follicles are critical culture steps for specifying competence of fully-grown oocytes (FGOs) for preimplantation development. A systematic comparison of transcriptomes of single oocytes having undergone different in vitro culture trajectories identifies genes normally upregulated during oocyte growth to be susceptible for mis-regulation during in vitro oogenesis. Many of such genes have been described as targets of Polycomb repressive complexes (PRCs). Deregulation of Polycomb repression therefore likely perturbs the accumulation of cytoplasmic factors and/or setting of chromatin states in FGOs that are required for embryonic development after fertilization. Conversely, in vitro derived oocytes often displayed failure of zygotic genome activation (ZGA) and abnormal acquisition of 5-hydroxymethylcytosine (5hmC) on maternal chromosomes after activation. In addition, subcellular delocalization of pyruvate dehydrogenase (PDH) and of STELLA were observed suggesting new molecular markers for defective oocyte development. Our study identifies epigenetic regulation at an early stage of oogenesis as crucial for developmental competence and suggests specific in vitro culture steps as targets for improving oocyte quality. HighlightsO_LISingle cell transcriptomics and functional assessment of oocyte development from pluripotent stem cells in culture in a stage-specific manner provides a comprehensive resource for comparisons to oogenesis in vivo. C_LIO_LICulture steps for growth and differentiation of reconstituted follicles are critical for defining embryonic competence of in vitro generated oocytes. C_LIO_LIZygotic genome activation failure and epigenetic impairment are hallmarks of in vitro-generated oocytes that fail to develop after activation or fertilization. C_LIO_LIComputational analysis of gene expression changes and chromatin modification patterns identifies specific gene sets that indicate that Polycomb mediated repression is vulnerable during in vitro folliculogenesis. C_LI

developmental biology↗