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Regulatory RNA binding proteins allow for specific control of gene expression in a very dynamic manner. In mammals ZFP36, formerly known as Tristetraprolin, controls the inflammatory response by binding to an AU-rich element located in the 3' untranslated region of its target mRNAs. The developping embryo relies on a population of primitive macrophages to ensure proper immunity. Although the role of zfp36 in adult immunity has been extensively studied, its expression in the developing immune system has been poorly documented. Here, we have used whole mount in situ hybridization with a 3' UTR specific probe to address the expression of zfp36 in developing Xenopus tropicalis embryos. We have shown that zfp36 is expressed in two distinct cellular populations. First, it is a new marker of primititive myeloid cells, being coexpressed with the myeloid marker mpo. Therefore this early expression may suggest a role for zfp36 in macrophage differentiation and activation. In addition, a second cell population was found to transiently express zfp36, but not mpo, along the fusing neural folds and may correspond to cells undergoing autophagy during neural tube closure.
Fig. 1. In situ hybridization for zfp36 using a 3’ UTR probe. Stages 20 and 26 X. tropicalis embryos were fixed and treated for ISH with a probe targeting the 3’ UTR of zfp36 mRNA. Ventral (vent.), lateral (lat.) and dorsal (dors.) views of representative embryos are shown as indicated.
Fig.2. zfp36 mRNA is expressed in two different cell populations in early X. tropicalis embryos. Double sequential in situ hybridization (A) revealed for zfp36 and zfp36 + mpo in stage 18 and stage 26 embryos. Vibratome section of stage 20 embryos (B) stained for zfp36 or mpo as indicated. The cut plane is shown on the right panel. (C) Mixed view of stages 19-21 embryos stained for zfp36 or mpo. Arrowhead, anterior neural fold staining for zfp36, open arrowhead, posterior labeling of neural fold by zfp36.
Fig. 3. Spatio-temporal expression of zfp36 mRNAs in X. tropicalis embryos. (A) RT-QPCR on total mRNA for the indicated stages of development for mpo and zfp36 mRNAs. Data are normalized to odc mRNAs. (B) Stage 14 to stage 24 X. tropicalis embryos are stained for zfp36 mRNAs and pictured form dorsal, ventral or frontal views. zfp36 mRNA is localized in the aVBI from stage 14 and along the neural fold starting from stage 19. (C) Stage 32 X. tropicalis embryos are stained for zfp36 mRNA with a sense or antisense probe as indicated. The dense ventral staining is indicated by an arrowhead.
zfp36 (ZFP36 ring finger protein) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 20, lateral view, anteriorleft, dorsal up.
zfp36 (ZFP36 ring finger protein) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 26, lateral view, anteriorleft, dorsal up.
zfp36 (ZFP36 ring finger protein) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 18, ventral view, anteriorleft.
zfp36 (ZFP36 ring finger protein) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 18, dorsal view, anteriorleft.
zfp36 (ZFP36 ring finger protein) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 32, lateral view, anteriorleft, dorsal up
