Handling Editor: Haruhiko Koseki
ABSTRACT
In mice, the entry of germ cells into meiosis crucially depends on the expression of stimulated by retinoic acid gene 8 (Stra8). Stra8 is expressed specifically in pre-meiotic germ cells of females and males, at fetal and postnatal stages, respectively, but the mechanistic details of its spatiotemporal regulation are yet to be defined. In particular, there has been considerable debate regarding whether retinoic acid is required, in vivo, to initiate Stra8 expression in the mouse fetal ovary. We show that the distinctive anterior-to-posterior pattern of Stra8 initiation, characteristic of germ cells in the fetal ovary, is faithfully recapitulated when 2.9 kb of the Stra8 promoter is used to drive eGFP expression. Using in vitro transfection assays of cutdown and mutant constructs, we identified two functional retinoic acid responsive elements (RAREs) within this 2.9 kb regulatory element. We also show that the transcription factor DMRT1 enhances Stra8 expression, but only in the presence of RA and the most proximal RARE. Finally, we used CRISPR/Cas9-mediated targeted mutation studies to demonstrate that both RAREs are required for optimal Stra8 expression levels in vivo.
Footnotes
Competing interests
The authors declare no competing or financial interests.
Author contributions
Conceptualization: C.-W.F., P.K., J.B.; Methodology: C.-W.F., G.B., K.C., J.B.; Formal analysis: C.-W.F., G.B.; Investigation: C.-W.F.; Resources: K.C.; Data curation: C.-W.F., G.B., C.M.S., F.K.M.C., J.B.; Writing - original draft: C.-W.F., C.M.S., J.B.; Writing - review & editing: C.-W.F., C.M.S., P.K., J.B.; Supervision: C.M.S., P.K., J.B.; Project administration: P.K., J.B.; Funding acquisition: P.K., J.B.
Funding
This work was supported by the National Health and Medical Research Council, Australia (APP1109502 to J.B.) and the Australian Research Council (DP140104059 to J.B. and DP110105459 to P.K. and J.B.).
Supplementary information
Supplementary information available online at https://dev.biologists.org/lookup/doi/10.1242/dev.194977.supplemental
- Received July 15, 2020.
- Accepted February 4, 2021.
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