Isolation of Intestinal Macrophage Subpopulations for High-Quality Total RNA Purification in Zebrafish

Yalén Del Río-Jay, Audrey Barthelaix, Cristian Reyes-Martínez, Christophe Duperray, Camila J. Solis-Cascante, Yessia Hidalgo, Patricia Luz-Crawford, Farida Djouad*, Carmen G. Feijoo*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


Intestinal macrophages have been poorly studied in fish, mainly due to the lack of specific molecular markers for their identification and isolation. To address this gap, using the zebrafish Tg(mpeg1:EGFP) transgenic line, we developed a fluorescence-activated cell sorting strategy (FACS) that allows us to isolate different intestinal macrophage subpopulations, based on GFP expression and morphological differences. Also, we achieved the purification of high-quality total RNA from each population to perform transcriptomic analysis. The complete strategy comprises three steps, including intestine dissection and tissue dissociation, the isolation of each intestinal macrophage population via FACS, and the extraction of total RNA. To be able to characterize molecularly different macrophage subpopulations and link them to their functional properties will allow us to unravel intestinal macrophage biology.

Original languageEnglish
Article number43
JournalMethods and Protocols
Issue number3
StatePublished - Jun 2024

Bibliographical note

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© 2024 by the authors.


  • intestinal macrophage
  • mRNA
  • zebrafish


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