Abstract
Holometabolous insects are predominantly motionless during metamorphosis, when no active feeding is observed and the body is enclosed in a hardened cuticle. These physiological properties as well as undergoing processes resemble embryogenesis, since at the pupal stage organs and systems of the imago are formed. Therefore, recapitulation of the embryonic expression program during metamorphosis could be hypothesized. To assess this hypothesis at the transcriptome level, we have performed a comprehensive analysis of the developmental datasets available in the public domain. Indeed, for most datasets, the pupal gene expression resembles the embryonic rather than the larval pattern, interrupting gradual changes in the transcriptome. Moreover, changes in the transcriptome profile during the pupa-to-imago transition are positively correlated with those at the embryo-to-larvae transition, suggesting that similar expression programs are activated. Gene sets that change their expression level during the larval stage and revert it to the embryonic-like state during the metamorphosis are enriched with genes associated with metabolism and development.
Similar content being viewed by others
Introduction
Hemi- and holometabolous insects differ in the magnitude of physiological and morphological changes during the metamorphosis. In hemimetabolous insects, embryogenesis typically ends up with an adult-like larva that further develops to the imago through sequential molts causing gradual shifts, with the wings and genitalia appearing during the adult molt. In holometabolous insects, the adult body plan is established at the prepupal and pupal periods, and larval organs and systems are de-differentiated and reorganized during the complete metamorphosis. This is usually accompanied by a more or less radical change in the habitat and feeding strategy. Larvae and adults of the same species do not share food resources, allowing the separation of growth and reproduction in time and space1.
Metamorphosis is believed to originate approximately 400 million years ago in the early Devonian, when Pterygota emerged, the insect flight was invented2, and complete metamorphosis evolved to support the ability to fly. Sequential molts require the whole body, including the wings, to be covered with the cuticle. It makes wings too heavy and almost no extant winged insects undergo molting during the imago stage, an exception being the short-living subimago of the mayfly that undergoes a full molting cycle to become the imago3.
During larval development some cells with latent embryonic potential are arrested and the differentiation process continues after the pupation4. These cells, initially forming so-called imaginal primordia, replace larval cells to form adult organs. The imaginal cells contribute little to the functioning of the larval organism and preserve pluripotency, similar to stem cells5. For example, in Papilio xuthus (Lepidoptera), a sophisticated orchestra of transcription factors that regulate the expression patterns of opsins, manifest only after the pupation to build the compound eye6. On the other hand, some organs undergo dedifferentiation followed by redifferentiation to the adult state. For example, in Drosophila (Diptera), syncytial alary muscles de-differentiate to mononuclear myoblasts prior to formation of the adult tissue7.
Differentiation of stem cells into mature tissues could reuse molecular mechanisms that drive the embryonic development, since the gain of new features is based on the upgrade of the existing ones8. Therefore, it could be hypothesized that the pupal gene expression program should resemble the embryonic one due to both differentiation de novo and redifferentiation. A study on the midge Polypedilum vanderplanki showed reversion of the transcriptional profile back to the embryonic stage during metamorphosis9. Here, we comprehensively analyze all insect developmental transcriptome datasets available in the public domain, with the aim to assess gene expression similarities between pupae and embryos.
Methods
Datasets
Developmental transcriptomic datasets with at least one sample originating from each of the four major stages (embryonic, larval, pupal, adult) in the holometabolous insect development were analyzed. The collection includes ten species from four orders (Diptera, Hymenoptera, Lepidoptera, Coleoptera). For Drosophila melanogaster, both RNA-seq and microarray datasets are available, and RNA-seq only for all other species, see Table 1 and Supplementary Tables S1–S9 for details. Specific timing of samples, source tissue and sex are shown in Supplementary Tables S1–S9 if provided in the original papers.
Mature females contain eggs; therefore, full-body transcriptomes have a strong signal from the eggs, yielding a high correlation of the female samples with the embryonic state. To avoid these confounding effects, the female samples were excluded from the analysis.
After the initial analysis, the Pieris rapae dataset10 was excluded due to insufficient data for the embryonic sample, the latter being an outlier with only 0.2M uniquely mapped reads, compared to about 13M reads for each other sample.
The Tribolium castaneum dataset comprises three replicates for each developmental stage. Two of the pupal samples had exactly the same set of raw reads; therefore, one copy was excluded.
RNAseq preprocessing
RNA sequencing reads were downloaded from the NCBI sequence read archive in the sra format, and fastq files containing reads were extracted. Low-quality reads that had low average nucleotide quality, shorter length than expected or high number of missing nucleotides were eliminated using the fastp tool23. Automatically detected adaptors (based on read overlapping analysis and built-in known adapter sequences from the fastp package) and low-quality regions at the end of the reads were trimmed.
For each organism, a reference transcriptome index was prepared by the Kallisto index tool24, see accessions of the published transcriptomes in Table 1. For both single-end and paired-end reads, mapping was performed using a pseudo alignment approach implemented in the Kallisto package. The FPKM (Fragments Per Kilobase Million) normalization was used for the downstream analysis.
Microarray dataset
Processed data containing log-transformed ratios between channels were downloaded from the NCBI GEO database (accession GSE3286) for three platforms separately. Probe IDs were converted into gene names using microarray specification information from NCBI GEO (related platforms are deposited under GPL2837, GPL2838 and GPL2840 accessions) and D. melanogaster gene information from the FlyBase database25.
Gene Ontology (GO) terms annotation
The D. melanogaster GO annotation was downloaded from the AmiGO 2 database26 for all three aspects (Biological Process, Molecular Function and Cellular Component).
InterProScan with the default parameters was used to predict GO terms for other species27. Protein sequences from the respective assemblies (see Table 1) were used as an input for InterProScan.
Genes were assumed to be related to development, if the Gene Ontology (GO) term “developmental process” (GO:0032502) or its descendant terms were predicted to be associated with the gene. These genes comprised the development-associated gene subset that was used further in the analysis. Genes predicted to have the GO term “metabolic process” (GO:0008152) or its descendants were regarded as metabolism-related. These genes comprised the metabolism-associated gene subset.
Across-stages similarity
Similarity between stages was measured by the Spearman correlation coefficient of the log-transformed FPKM values.
Random sampling
To assess the influence of particular gene subsets on the observed transcriptome characteristics, random sampling was performed. At that, gene sets of the same size as the selected gene set were randomly sampled several times. For each random gene set the desired metric was calculated. The obtained distribution of the possible metric values was used as a reference distribution to estimate the p-value or quantile of the observed data.
Gene profile clustering
For datasets with data available for four main stages only, there are 27 possible patterns of gene expression (it could increase, decrease or remain the same during each of three transitions between stages). A gene was assigned to the pattern with which it had the highest correlation. Thus, for such datasets, genes were divided into 27 clusters. We were specifically interested in two clusters corresponding to the zigzag pattern of gene expression across the development, where the transcriptome profile reverts back to the embryonic state during metamorphosis.
The transcriptome datasets with more than four time points were hierarchically clustered with the Spearman correlation coefficient as the distance metric. The hierarchy algorithm from the scipy package was used28.
GO term enrichment analysis
Python package goatools was used to identify significantly enriched GO terms29 using the default parameters with an adjusted p-value threshold equal to 0.05. The set of all genes with positive estimated expression values in the corresponding dataset was used as the reference set for GO enrichment analysis.
Visualization
Python 3.7, matplotlib and seaborn packages were used for the visualization. Plots for the semantic analysis of the GO enrichment results were generated using the REVIGO tool30 and the R ggplot package.
Results and discussion
Intra-species comparison
Intra-species comparisons across developmental stages allowed us to compare the transcriptome profiles at several distinct time points. A monotonic development results in gene expression patterns at each particular stage being closest to the immediately previous and following developmental stages, yielding a decrease of the similarity with the increase of the time interval between the time points. Indeed, this behavior is observed for the embryonic and larval stages. The correlation coefficient decreases for relatively more distant stages, as seen in the pairwise correlation heatmap for the detailed D. melanogaster dataset (Fig. 1). In that case, high Spearman correlation coefficients are concentrated near the diagonal.
However, this monotonic development is interrupted during the pupation suggesting drastic changes in the transcriptome profile. Gene expression levels at early prepupal and pupal stages are closer to the embryonic profiles rather than to the larval ones. It suggests some crucial event to happen during prepupal stages that will drive development towards formation of the adult body. An example of such an event could be the loss of the juvenile hormone that is thought to reactivate morphogenesis31. Moreover, in Manduca sexta, the level of the juvenile hormone decreases significantly before entering the prepupal stages (to trigger cell proliferation) with a narrow burst of the hormone titre during the prepupal development (to prevent precocious adult development)32,33.
The monotonic development is restarted at some point during the metamorphosis, extending to the adult stages, so that high correlation coefficients are again observed close to the diagonal of the matrix.
Datasets from several other insect species, though less detailed, demonstrate the same overall pattern, with pupal transcriptomes being more similar to the embryonic ones than to the larval or adult ones. Sample heatmaps for moth Ostrinia furnacalis and beetle T. castaneum are presented in Fig. 2 (left). Heatmaps for other datasets see in Supplementary Fig. S1.
The effect of increased similarity between embryo and pupa compared to the embryo-larvae similarity is observed in several more datasets (Fig. 3a, left); therefore, it is not restricted to the Drosophila genus or the Diptera order. However, for some species pupae do not resemble embryos, an example being M. sexta (Fig. 3a, gray line). This could be explained by the fact that the M. sexta samples were collected from the whole body for early developmental stages and from several specific tissues for later stages. This makes direct transcriptome comparisons less reliable, since differences could be tissue-specific regardless of the developmental stage. In other cases, a possible explanation is that early or late pupae have been collected, closer to the adjacent larval or adult stages, respectively.
The latter explanation is supported by three D. melanogaster microarray datasets (Fig. 3b). Indeed, while middle pupal stages are more similar to the embryonic stages, late pupae are more similar to adults.
D. melanogaster datasets fall into two groups depending on the technology used to generate the source data. The RNA sequencing datasets (blue and green lines in Fig. 3b) demonstrate a monotonic decrease in similarity with the embryo in the course of development.
Functional subsets of genes
To understand the molecular basis of the observed pattern, functional subsets of genes were considered. During metamorphosis, tissues are reorganized or even developed de novo from stem cells, like in embryogenesis. From the lifestyle point of view, the pupa also resembles the embryo since it is motionless and lacks active feeding. Therefore, genes with Gene Ontology (GO) terms related to “developmental process” (GO:0032502) and “metabolic process” (GO:0008152) were tested to account for the observed effect.
For D. melanogaster RNA-seq datasets, the analysis of subsets yields largely similar results. For two detailed microarray datasets, genes associated with metabolic processes demonstrate higher correlation between the embryonic and pupal samples. However, the range of the interval between quartiles makes the observation unreliable.
Similarity between embryo and pupa is higher, when calculated based on gene subsets, rather than the entire dataset for the O. furnacalis (Fig. 2, middle top). To test the statistical significance of the finding, random sampling was performed (see Methods). A high quantile of the observed correlation coefficient is expected for gene subsets strongly influencing the effect. On the contrary, gene subsets that have expression patterns following the average trend would have quantile values close to one-half. High correlation between embryonic and pupal transcriptomes in O. furnacalis data is supported by quantile values (Fig. 2, right top), suggesting that both development and metabolism-associated genes are collinearly expressed during these stages.
On the other hand, in T. castaneum, the expression of development-associated genes does not follow this trend (Fig. 2, middle bottom). At that, it should be noted that D. melanogaster is the only analyzed species with verified GO terms annotation from a dedicated database, while the GO annotation for other species is predicted using InterProScan. Moreover, only 1% of all proteins are annotated as associated with GO:0032502 (“developmental process”), leading to noisy results (Fig. 3a, middle). On the contrary, as many as 40% of proteins in each species are assigned with GO:0008152 (“metabolic process”) and hence the correlation coefficients naturally are close to those obtained using the complete datasets (Fig. 3a, right). However, for both types of subsets, there are species with an enhanced effect.
The reversion of the transcriptome pattern
As described above, the transcriptome profile of holometabolous insects tends to revert to the embryonic state during metamorphosis. This can be seen not only from direct comparison of transcriptomes on several developmental stages, but from the analysis of transcriptome changes during transitions between adjacent stages. Indeed, in some cases, changes in gene expression that happen at the pupa-to-imago transition recapitulate the egg-to-larva transition. For example, the left part of Fig. 4 shows changes in gene expression for the O. furnacalis and T. castaneum datasets. A positive correlation between fold-change values is observed in 75% of the datasets. 40% of them have a correlation coefficient higher than 0.1, suggesting that the metamorphosis developmental program that drives (re-)formation of tissues and organs indeed dynamically recapitulates the embryo differentiation.
The effect of synchronized changes in expression patterns during embryo and pupal eclosion could be explained by monotonous processes occurring in the complete course of development. However, it could not be the main explanation, since in pairwise comparisons gradual changes are not observed and the pupal transcriptome is more similar to the embryonic rather than larval one.
For O. furnacalis, the development subset demonstrates higher correlation between fold-changes (middle top of the Fig. 4). Downsampling p-value supports this observation, since only 1% of random gene subsets show higher correlation. However, due to the low number of genes with predicted links to development, a significant effect is seen in only five datasets with positive correlation (Fig. 5, left).
Higher than average fold-change correlation for metabolism-associated genes is observed across more datasets (Fig. 5, right), and it is statistically significant for most of the species. Therefore, metabolic genes could partially drive the recapitulation.
Genes that drive recapitulation should have a zigzag-like pattern of gene expression during development. To identify such genes, clustering of the expression profiles was performed. Datasets with one time point measured for each of the main stages (embryo, larva, pupa and imago) are scored by a correlation coefficient with one of the possible 27 artificial trajectories (with up/same/down steps).
More detailed datasets were clustered hierarchically (see Methods). The M. sexta dataset was not considered at this step since 72% of its samples correspond to the larval stage and therefore the cluster diversity is dominated by gene expression changes during the larval development. For other datasets, clusters with the pattern of similar expression in the embryonic and pupal samples (zigzag-like pattern) were selected for further analysis (Supplementary Fig. S2). The set of genes with expression that increases while entering the larval stage and then decreases after the pupation is enriched with several classes of metabolism and development-associated terms (Fig. 6).
Terms similar to purine containing compound metabolic process (GO:0072521), ribose phosphate metabolic process (GO:0019693), ATP metabolic process (GO:0046034), electron transport chain (GO:0022900), proton transmembrane transport (GO:1902600) and oxidation–reduction process (GO:0055114) in the semantic space for biological processes category suggest a high rate of energy-generation and consumption during both embryogenesis34 and pupa maturation to build organs and tissues.
The enriched peptidase activity (GO:0008233) molecular function could be involved in several processes. For example, matrix metalloproteinases regulate trachea and intestinal development in embryo and pupal morphogenesis of T. castaneum beetle35; caspases, along with other proteases are key players in the induced cell death during metamorphosis, it is essential for remodeling of the larval tissue36. Peptidases also balance proliferation, being, in particular, crucial players in development of the tracheal system37 or neuroblasts38 in D. melanogaster.
Cell–cell adhesion (GO:0098609), multicellular organismal process (GO:0032501) and anatomical structure development (GO:0048856) terms are frequent among active genes during the embryo and pupal stages. In particular, these terms were enriched in the respective cluster in the D. melanogaster RNA-seq dataset12 (Fig. 7a and b, respectively). The correlation heatmap for this cluster features a distinct diagonal, suggesting each stage is similar to the ones that are close in time (Fig. 7c). However, there is a prominent diagonal in the embryo-pupae submatrix suggesting involvement of similar processes.
Conclusions
Several datasets indeed show an increased similarity between the embryonic and pupal stages on the gene expression level when compared with the embryo-larvae transcriptome pairs. Sets of genes changing their expression level during the larval stage and returning to the embryonic state during the metamorphosis are enriched with genes related to energy metabolism and multicellular organism development.
Gene expression changes at transition from the embryonic to the larval stage for some datasets are correlated with changes between pupa and imago, suggesting similarity of transcriptional programs during embryonic development and pupal maturation. Separate analysis of metabolism-associated genes and genes related to the development enhances the observed effect for most datasets.
However, some datasets do not follow the pattern of embryonic expression recapitulation during morphogenesis. This might be due to the timing of collected pupal stages, as early pupae naturally resemble late larvae, while late pupae are similar to imagoes. Still, we consider the hypothesis to be tentatively confirmed and submit it for detailed experimental validation.
Two main opinions regarding the origin of metamorphosis origin in evolution are discussed in the literature. The Hinton hypothesis proposes the pupa to arise from the final nymphal instar of a hemimetabolous ancestor39. An alternative hypothesis suggests the larva of holometabolous insects represent an arrest phase of embryonic development, therefore metamorphosis is a continuation of embryogenesis40, a suggestion traced back to William Harvey41. In that case the pupa would correspond to all nymphal instars of hemimetabolous insects. The latter hypothesis is not supported by our observations, since it implies gradual development with a stalled larval stage without drastic changes during the prepupal and pupal period, contrary to our findings. However, although it is clearly a derived trait, as in most other Diptera the pupa is motionless, it might mean that some parts of the larval regulatory network are still active. At that, one might even expect that a detailed transcriptome analysis with good temporal resolution in a sufficient number of diverse species would demonstrate that both explanations are true to some extent, with the balance between continuous, monotonic developmental program (implied by the Harvey hypothesis) and considerable change in the transcriptome (as in the Hinton hypothesis) with partial recapitulation of the early embryonic transcription program (as observed here) would shift in different species and for different functional subsystems.
Data availability
The datasets analyzed in this study are available in the NCBI Gene Expression Omnibus under the accession GSE3286 — D. melanogaster microarray dataset13 and in the Sequence Read Archive under accessions SRA00936412, SRA01217311, SRP05302214, SRP04584615, SRP22012016, SRP05775017, SRP05901210, SRP07085418, SRP00637119, SRP04723620, SRP06525521 and DRP0024059 — RNA sequencing datasets.
References
Belles, X. Origin and evolution of insect metamorphosis. Encyclopedia of Life Sciences (ELS). 1–11 (John Wiley and Sons, Ltd, Chichester, 2011).
Misof, B. et al. Phylogenomics resolves the timing and pattern of insect evolution. Science 346, 763–767 (2014).
Edmunds, G. F. & McCafferty, W. P. The mayfly subimago. Annu. Rev. Entomol. 33, 509–527 (1988).
Truman, J. W. & Riddiford, L. M. Endocrine insights into the evolution of metamorphosis in insects. Annu. Rev. Entomol. 47, 467–500 (2002).
McClure, K. D. & Schubiger, G. Transdetermination: Drosophila imaginal disc cells exhibit stem cell-like potency. Int. J. Biochem. Cell Biol. 39, 1105–1118 (2007).
Arikawa, K., Iwanaga, T., Wakakuwa, M. & Kinoshita, M. Unique temporal expression of triplicated long-wavelength opsins in developing butterfly eyes. Front. Neural Circuits 11, 96 (2017).
Schaub, C., März, J., Reim, I. & Frasch, M. Org-1-dependent lineage reprogramming generates the ventral longitudinal musculature of the Drosophila heart. Curr. Biol. 25, 488–494 (2015).
Koonin, E. V. The Logic of Chance: The Nature and Origin of Biological Evolution. (FT Press, 2011).
Mazin, P. V. et al. Cooption of heat shock regulatory system for anhydrobiosis in the sleeping chironomid Polypedilum vanderplanki. Proc. Natl. Acad. Sci. U. S. A. 115, E2477–E2486 (2018).
Qi, L. et al. De novo assembly and developmental transcriptome analysis of the small white butterfly pieris rapae. PLoS ONE 11, e0159258 (2016).
Daines, B. et al. The Drosophila melanogaster transcriptome by paired-end RNA sequencing. Genome Res. 21, 315–324 (2011).
Graveley, B. R. et al. The developmental transcriptome of Drosophila melanogaster. Nature 471, 473–479 (2011).
Arbeitman, M. N. et al. Gene expression during the life cycle of Drosophila melanogaster. Science 297, 2270–2275 (2002).
Wu, Z. et al. Discovery of chemosensory genes in the oriental fruit fly, Bactrocera dorsalis. PLoS ONE 10, e0129794 (2015).
Sim, S. B., Calla, B., Hall, B., DeRego, T. & Geib, S. M. Reconstructing a comprehensive transcriptome assembly of a white-pupal translocated strain of the pest fruit fly Bactrocera cucurbitae. Gigascience 4, 14 (2015).
Wei, D. et al. Genome-wide gene expression profiling of the melon fly, Zeugodacus cucurbitae, during thirteen life stages. Sci Data 7, 45 (2020).
Jones, B. M., Wcislo, W. T. & Robinson, G. E. Developmental transcriptome for a facultatively eusocial bee. Megalopta genalis. G3(5), 2127–2135 (2015).
Zhang, T., He, K. & Wang, Z. Transcriptome comparison analysis of ostrinia furnacalis in four developmental stages. Sci. Rep. 6, 35008 (2016).
He, W. et al. Developmental and insecticide-resistant insights from the de novo assembled transcriptome of the diamondback moth, Plutella xylostella. Genomics 99, 169–177 (2012).
Cao, X. & Jiang, H. An analysis of 67 RNA-seq datasets from various tissues at different stages of a model insect, Manduca sexta.. BMC Genomics 18, 796 (2017).
Perkin, L., Elpidina, E. N. & Oppert, B. Expression patterns of cysteine peptidase genes across the Tribolium castaneum life cycle provide clues to biological function. PeerJ 4, e1581 (2016).
Gusev, O. et al. Comparative genome sequencing reveals genomic signature of extreme desiccation tolerance in the anhydrobiotic midge. Nat. Commun. 5, 4784 (2014).
Chen, S., Zhou, Y., Chen, Y. & Gu, J. fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics 34, i884–i890 (2018).
Bray, N. L., Pimentel, H., Melsted, P. & Pachter, L. Near-optimal probabilistic RNA-seq quantification. Nat. Biotechnol. 34, 525–527 (2016).
Gramates, L. S. et al. FlyBase: a guided tour of highlighted features. Genetics 220(4), iyac035 (2022).
Gene Ontology Consortium. The gene ontology resource: Enriching a gold mine. Nucleic Acids Res. 49, D325–D334 (2021).
Blum, M. et al. The InterPro protein families and domains database: 20 years on. Nucleic Acids Res. 49, D344–D354 (2021).
Virtanen, P. et al. SciPy 1.0: Fundamental algorithms for scientific computing in Python. Nat. Methods 17, 261–272 (2020).
Klopfenstein, D. V. et al. GOATOOLS: A Python library for Gene Ontology analyses. Sci. Rep. 8, 10872 (2018).
Supek, F., Bošnjak, M., Škunca, N. & Šmuc, T. REVIGO summarizes and visualizes long lists of gene ontology terms. PLoS ONE 6, e21800 (2011).
Truman, J. W. & Riddiford, L. M. The morphostatic actions of juvenile hormone. Insect Biochem. Mol. Biol. 37, 761–770 (2007).
Riddiford, L. M. Cellular and Molecular Actions of Juvenile Hormone I. General Considerations and Premetamorphic Actions. in Advances in Insect Physiology (ed. Evans, P. D.) vol. 24 213–274 (Academic Press, 1994).
Konopova, B., Smykal, V. & Jindra, M. Common and distinct roles of juvenile hormone signaling genes in metamorphosis of holometabolous and hemimetabolous insects. PLoS ONE 6, e28728 (2011).
Gándara, L. & Wappner, P. Metabo-Devo: A metabolic perspective of development. Mech. Dev. 154, 12–23 (2018).
Knorr, E., Schmidtberg, H., Vilcinskas, A. & Altincicek, B. MMPs regulate both development and immunity in the tribolium model insect. PLoS ONE 4, e4751 (2009).
Tettamanti, G. & Casartelli, M. Cell death during complete metamorphosis. Philos. Trans. R. Soc. Lond. B Biol. Sci. 374, 20190065 (2019).
Baer, M. M. et al. The role of apoptosis in shaping the tracheal system in the Drosophila embryo. Mech. Dev. 127, 28–35 (2010).
Harding, K. & White, K. Decoupling developmental apoptosis and neuroblast proliferation in Drosophila. Dev. Biol. 456, 17–24 (2019).
Hinton, H. E. The origin and function of the pupal stage. Proc. R. Entomol. Soc. Lond. 38, 77–85 (2009).
Truman, J. W. & Riddiford, L. M. The origins of insect metamorphosis. Nature 401, 447–452 (1999).
Erezyilmaz, D. F. Imperfect eggs and oviform nymphs: A history of ideas about the origins of insect metamorphosis. Integr. Comp. Biol. 46, 795–807 (2006).
Acknowledgements
This study was supported by grants 17-00-00180 (transcriptome analysis) and 20-54-81007 (data collection and functional analysis) from the Russian Foundation for Basic Research. We are grateful to Georgii Bazykin, Ekaterina Khrameeva, Andrei Mironov, and Mikhail Moldovan for useful discussions.
Author information
Authors and Affiliations
Contributions
O.A.M.*—Data curation, Formal analysis, Investigation, Methodology, Software, Validation, Visualization, Writing – original draft. G.M.S.—Conceptualization, Funding acquisition, Methodology, Project administration, Resources, Supervision, Validation, Writing – review & editing.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
About this article
Cite this article
Ozerova, A.M., Gelfand, M.S. Recapitulation of the embryonic transcriptional program in holometabolous insect pupae. Sci Rep 12, 17570 (2022). https://doi.org/10.1038/s41598-022-22188-y
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41598-022-22188-y
- Springer Nature Limited