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Differential gene expression and hallmarks of stemness in epithelial cells of the developing rat epididymis

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Abstract

Epididymal development can be subdivided into three phases: undifferentiated, a period of differentiation, and expansion. The objectives of this study were (1) to assess gene expression profiles in epididymides, (2) predict signaling pathways, and (3) develop a novel 3D cell culture method to assess the regulation of epididymal development in vitro. Microarray analyses indicate that the largest changes in differential gene expression occurred between the 7- to 18-day period, in which 1452 genes were differentially expressed, while 671 differentially expressed genes were noted between days 18 and 28, and there were 560 differentially expressed genes between days 28 and 60. Multiple signaling pathways were predicted at different phases of development. Pathway associations indicated that in epididymides of 7- to 18-day old rats, there was a significant association of regulated genes implicated in stem cells, estrogens, thyroid hormones, and kidney development, while androgen- and estrogen-related pathways were enriched at other phases of development. Organoids were derived from CD49f + columnar cells from 7-day old rats, while no organoids developed from CD49f− cells. Cells cultured in an epididymal basal cell organoid medium versus a commercial kidney differentiation medium supplemented with DHT revealed that irrespective of the culture medium, cells within differentiating organoids expressed p63, AQP9, and V-ATPase after 14 days of culture. The commercial kidney medium resulted in an increase in the number of organoids positive for p63, AQP9, and V-ATPase. Together, these data indicate that columnar cells represent an epididymal stem/progenitor cell population.

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Data availability

The gene expression data from transcriptomic analyses have been deposited in the National Center for Biotechnology Information’s Gene Expression Omnibus (GEO, http://www.ncbi.nlm.nih.gov/geo/) and are accessible through GEO accession number GSE193653. Other data are available from the corresponding author.

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Acknowledgements

The assistance of T. El Belaidi and S. Pinto (INRS) is appreciated. J. Tremblay (INRS) is thanked for his assistance with confocal microscopy and flow cytometry.

Funding

The study was funded by grants to DGC from the Canadian Institutes for Health Research (84576), Natural Sciences and Engineering Research Council (155065–06), and Canada Research Chairs Program. LP is the recipient of a studentship from the Fonds de Recherches du Québec-Nature et Technologie -Réseau Québécois en Reproduction.

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Author notes

  1. Julie Dufresne and Mary Gregory are both contributed equally.

Authors and Affiliations

  1. Laboratory for Reproductive Toxicology, INRS-Centre Armand-Frappier Santé Biotechnologie, Université du Québec, 245 boul. Des Prairies, Laval, QC, H7V 3B7, Canada

    Julie Dufresne, Mary Gregory, Laurie Pinel & Daniel G. Cyr

  2. Department of Anatomy and Cell Biology, McGill University, Montreal, QC, Canada

    Daniel G. Cyr

  3. Department of Obstetrics, Gynecology, and Reproduction, Laval University, Québec, QC, Canada

    Daniel G. Cyr

Authors
  1. Julie Dufresne
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  2. Mary Gregory
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  3. Laurie Pinel
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  4. Daniel G. Cyr
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Corresponding author

Correspondence to Daniel G. Cyr.

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Ethics approval

All the animal protocols used in this study were approved by the Institut National de la recherche Scientifique (INRS) University Animal Care Committee according to the guidelines of the Canadian Council on Animal Care.

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Dufresne, J., Gregory, M., Pinel, L. et al. Differential gene expression and hallmarks of stemness in epithelial cells of the developing rat epididymis. Cell Tissue Res 389, 327–349 (2022). https://doi.org/10.1007/s00441-022-03634-9

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  • DOI: https://doi.org/10.1007/s00441-022-03634-9

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