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We thank the UPF Flow Cytometry Core Facility for their constant kind assistance. We thank the CRG Advanced Light Microscopy Unit for their help in acquisition of confocal videos and images. We thank David Booth for his advice on the transfection protocol and generous sharing of carrier plasmid DNA. We thank Meritxell Anto ' for technical support. We thank Omaya Dudin and Nu ' ria Ros-Rocher for their support in analyzing microscopy images and the MulticellGenome laboratory for fruitful discussions. This work was funded by the Gordon and Betty Moore Foundation's Marine Microbiology Initiative for establishing Emerging Model Systems Grant Number: 4973.01.

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November 15, 2021
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Report Stable transfection in protist Corallochytrium limacisporum identifies novel cellular features among unicellular animals relatives

Publicated to:Current Biology. 31 (18): 4104-+ - 2021-09-27 31(18), DOI: 10.1016/j.cub.2021.06.061

Authors: Kozyczkowska, A; Najle, SR; Ocana-Pallares, E; Areste, C; Shabardina, V; Ara, PS; Ruiz-Trillo, I; Casacuberta, E

Affiliations

Barcelona Inst Sci & Technol GIST, Ctr Genom Regulat CRG, Barcelona 08003, Catalonia, Spain - Author
ICREA, Passeig Lluis Companys 23, Barcelona 08010, Catalonia, Spain - Author
IDIBAPS, IIBB CSIC, Dept Cell Death & Proliferat, Barcelona 08036, Spain - Author
Univ Barcelona, Dept Genet Microbiol & Estadist, Av Diagonal 645, Barcelona 08028, Catalonia, Spain - Author
Univ Pompeu Fabra, CSIC, Inst Biol Evolut, Passeig Maritim de la Barceloneta 37-49, Barcelona 08003, Catalonia, Spain - Author
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Abstract

The evolutionary path from protists to multicellular animals remains a mystery. Recent work on the genomes of several unicellular relatives of animals has shaped our understanding of the genetic changes that may have occurred in this transition.1-3 However, the specific cellular modifications that took place to accommodate these changes remain unclear. To address this, we need to compare metazoan cells with those of their extant relatives, which are choanoflagellates, filastereans, ichthyosporeans, and corallochytreans/pluriformeans. Interestingly, these lineages display a range of developmental patterns potentially homologous to animal ones. Genetic tools have already been established in three of those lineages.4-' However, there are no genetic tools available for Corallochytrea. We here report the development of stable transfection in the corallochytrean Corallochytrium limacisporum. Using these tools, we discern previously unknown biological features of C. limacisporum. In particular, we identify two different paths for cell division-binary fission and coenocytic growth-that reveal a non-linear life cycle. Additionally, we found that C. limacisporum is binucleate for most of its life cycle, and that, contrary to what happens in most eukaryotes, nuclear division is decoupled from cellular division. Moreover, its actin cytoskeleton shares characteristics with both fungal and animal cells. The establishment of these tools in C. limacisporum fills an important gap in the unicellular relatives of animals, opening up new avenues of research to elucidate the specific cellular changes that occurred in the evolution of animals.

Keywords

BiologyCorallochytrea, unicellular holozoa, binary fission, genetic tools, stable transfection, binucleated cells, coenocyte, non-linear life cycle, decoupled karyokinesis and cytokinesisDnaEvolutionOriginPhylogenyReveal

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal Current Biology due to its progression and the good impact it has achieved in recent years, according to the agency WoS (JCR), it has become a reference in its field. In the year of publication of the work, 2021, it was in position 3/94, thus managing to position itself as a Q1 (Primer Cuartil), in the category Biology. Notably, the journal is positioned above the 90th percentile.

From a relative perspective, and based on the normalized impact indicator calculated from the Field Citation Ratio (FCR) of the Dimensions source, it yields a value of: 5.58, which indicates that, compared to works in the same discipline and in the same year of publication, it ranks as a work cited above average. (source consulted: Dimensions Jul 2025)

Specifically, and according to different indexing agencies, this work has accumulated citations as of 2025-07-06, the following number of citations:

  • WoS: 11
  • Europe PMC: 14

Impact and social visibility

From the perspective of influence or social adoption, and based on metrics associated with mentions and interactions provided by agencies specializing in calculating the so-called "Alternative or Social Metrics," we can highlight as of 2025-07-06:

  • The use, from an academic perspective evidenced by the Altmetric agency indicator referring to aggregations made by the personal bibliographic manager Mendeley, gives us a total of: 33.
  • The use of this contribution in bookmarks, code forks, additions to favorite lists for recurrent reading, as well as general views, indicates that someone is using the publication as a basis for their current work. This may be a notable indicator of future more formal and academic citations. This claim is supported by the result of the "Capture" indicator, which yields a total of: 37 (PlumX).

With a more dissemination-oriented intent and targeting more general audiences, we can observe other more global scores such as:

  • The Total Score from Altmetric: 9.5.
  • The number of mentions on the social network X (formerly Twitter): 11 (Altmetric).
  • The number of mentions on Wikipedia: 1 (Altmetric).

It is essential to present evidence supporting full alignment with institutional principles and guidelines on Open Science and the Conservation and Dissemination of Intellectual Heritage. A clear example of this is:

  • The work has been submitted to a journal whose editorial policy allows open Open Access publication.