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Single-cell RNA sequencing of mid-to-late stage spider embryos: new insights into spider development
Uppsala University, Disciplinary Domain of Science and Technology, Earth Sciences, Department of Earth Sciences, Palaeobiology.ORCID iD: 0000-0002-9472-4928
Uppsala University, Disciplinary Domain of Science and Technology, Earth Sciences, Department of Earth Sciences, Palaeobiology.ORCID iD: 0000-0001-9007-4369
Uppsala University, Disciplinary Domain of Science and Technology, Earth Sciences, Department of Earth Sciences, Palaeobiology.ORCID iD: 0000-0002-4026-4129
2024 (English)In: BMC Genomics, E-ISSN 1471-2164, Vol. 25, article id 150Article in journal (Refereed) Published
Abstract [en]

Background

The common house spider Parasteatoda tepidariorum represents an emerging new model organism of arthropod evolutionary and developmental (EvoDevo) studies. Recent technical advances have resulted in the first single-cell sequencing (SCS) data on this species allowing deeper insights to be gained into its early development, but mid-to-late stage embryos were not included in these pioneering studies.

Results

Therefore, we performed SCS on mid-to-late stage embryos of Parasteatoda and characterized resulting cell clusters by means of in-silico analysis (comparison of key markers of each cluster with previously published information on these genes). In-silico prediction of the nature of each cluster was then tested/verified by means of additional in-situ hybridization experiments with additional markers of each cluster.

Conclusions

Our data show that SCS data reliably group cells with similar genetic fingerprints into more or less distinct clusters, and thus allows identification of developing cell types on a broader level, such as the distinction of ectodermal, mesodermal and endodermal cell lineages, as well as the identification of distinct developing tissues such as subtypes of nervous tissue cells, the developing heart, or the ventral sulcus (VS). In comparison with recent other SCS studies on the same species, our data represent later developmental stages, and thus provide insights into different stages of developing cell types and tissues such as differentiating neurons and the VS that are only present at these later stages.

Place, publisher, year, edition, pages
BioMed Central (BMC), 2024. Vol. 25, article id 150
Keywords [en]
Single-cell sequencing, Spider development, Nervous system, Genetic fingerprint, Parasteatoda tepidariorum
National Category
Developmental Biology Bioinformatics and Computational Biology
Identifiers
URN: urn:nbn:se:uu:diva-523429DOI: 10.1186/s12864-023-09898-xISI: 001281686900002PubMedID: 38326752Scopus ID: 2-s2.0-85184707233OAI: oai:DiVA.org:uu-523429DiVA, id: diva2:1838791
Part of project
SNIC 2.0: Swedish National Infrastructure for Computing, Swedish Research Council
Funder
Uppsala UniversityEU, Horizon 2020, 766053Swedish National Infrastructure for Computing (SNIC)UPPMAXSwedish Research Council, 2018‑05973Available from: 2024-02-19 Created: 2024-02-19 Last updated: 2025-02-17Bibliographically approved
In thesis
1. Single-cell RNA sequencing provides novel insights into spider development and represents an innovative alternative to study the evolution and development of panarthropods
Open this publication in new window or tab >>Single-cell RNA sequencing provides novel insights into spider development and represents an innovative alternative to study the evolution and development of panarthropods
2024 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Panarthropoda is a monophyletic group of invertebrate animals with a segmented body, paired appendages, dorsal brain, and ventral nerve cords. In order to study the mechanisms underpining their evolution, I study the genetic factors that drive their development. A typical research strategy is the candidate gene approach, in which orthologs of genes from a well established model organisms such as the fruit fly Drosophila melanogaster are studied in other more or less related species for comparison.

Recently developed single-cell RNA sequencing technologies allow the profiling of gene expression on the level of individual cells, and thus provide a much more detailed insight into gene expression.

In Paper-I, I applied the candidate gene approach to study the potential role of two transcription factors, called tiptop/teashirt and spalt, as trunk-selectors in panarthropods.

In Paper-II, I applied single-cell RNA sequencing to obtain the transcriptome of embryonic cells from spiders at mid-to-late stage in development. This generated a gene expression/gene-cell matrix that I analyzed to define the identity of cell clusters.

In Paper-III, I present an improved SCS data analysis based on the data presented in Paper-II. This revealed a number of new cell clusters including a cluster that is characterized by known eye-developmental genes, genes that have previously not been identified as eye-developmental genes, and hitherto un-investigated genes. My in-situ hybridization analyis shows that these genes are potential novel factors of eye development in the spider.

This work constitutes a successful example of the advantages of applying scRNA-seq in the study of panarthropod evolution and development.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2024. p. 55
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 2370
Keywords
panarthropod, spider embryo, single-cell RNA sequencing, EvoDevo, candidate gene approach, gene expression, cluster marker
National Category
Natural Sciences Biological Sciences
Research subject
Biology with specialization in Molecular Biology; Earth Science with specialization in Historical Geology and Palaeontology
Identifiers
urn:nbn:se:uu:diva-523957 (URN)978-91-513-2051-9 (ISBN)
Public defence
2024-04-22, Axel Hambergsalen, Uppsala, 08:00 (English)
Opponent
Supervisors
Funder
EU, Horizon 2020, 766053
Available from: 2024-03-21 Created: 2024-02-27 Last updated: 2024-04-15

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Medina-Jiménez, Brenda IreneBudd, Graham E.Janssen, Ralf

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