Abstract
Although the roles of the Hippo pathway in organogenesis and tumorigenesis have been well studied in multiple organs, its role in sperm maturation and male fertility has not been investigated. The initial segment (IS) of the epididymis plays a critical role in sperm maturation. IS differentiation is governed by ERK1/2, but the mechanisms of ERK1/2 activation in IS are not fully understood. Here we show that double knockout (dKO) of mammalian sterile 20-like kinases 1 and 2 (Mst1 and Mst2), homologs of Hippo in Drosophila, in the epididymal epithelium led to male infertility in mice. Sperm in the cauda epididymides of mutant mice were immotile with flagellar angulation and severely disorganized structures. Loss of Mst1/2 activated YAP and increased proliferation and cell death in all the segments of epididymis. The mutant mice showed substantially suppressed MEK/ERK signaling in the IS and failed IS differentiation. Deletion of Yap restored the reduced MEK/ERK signaling, and partially rescued the defective IS differentiation and fertility in Mst1/2 dKO mice. Our results demonstrate that YAP inhibits the MEK/ERK pathway in IS epithelial cells, and MST1/2 control IS differentiation and fertility at least partially by repressing YAP. Taken together, the Hippo pathway is essential for sperm maturation and male fertility.
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Acknowledgements
We thank Dr Sylvie Breton (Massachusetts General Hospital) for providing the AQP9 and B1-ATPase antibodies, Dr Yu Huang (The Chinese University of Hong Kong) for the financial support for RNA-Seq, Dr Wen-Liang Zhou (Sun Yat-Sen University) for PC-1 cells, Dr Xiangjian Zheng (Tianjin Medical University) for the help with X-gal staining, and Sharon Y.C. Ruan (The Hong Kong Polytechnic University) for helpful discussions.
We are grateful to Josie Lai and Jean Kung (The Chinese University of Hong Kong) for technical assistance with the electronic microscopy and Corinna Au (The Chinese University of Hong Kong) for assistance with histology.
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41418_2020_544_MOESM6_ESM.tif
Supplemental Figure 5. Comparison of epithelial height of region I of initial segment between WT and Mst1/2 dKO mice at 8 weeks of age.
41418_2020_544_MOESM7_ESM.tif
Supplemental Figure 6. Phosphorylation levels of the kinases upstream of ERK1/2 and protein levels of MST1, MST2 and YAP in the initial segment and cauda epididymis between WT and Mst1/2 dKO mice.
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Supplemental Figure 7. H&E-stained paraffin-embedded sections of caput epididymides from WT and Mst1/2 dKO mice at 4 and 8 weeks of age.
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Supplemental Figure 9. Comparison of epithelial height of initial segment between WT and Mst1/Mst2/Yap tKO mice at 8 weeks of age.                    Supplemental Figure 10. The top GO terms associated with molecular functions of differently expressed genes between the initial segments of WT and Mst1/Mst2/Yap tKO mice.
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Meng, C., Tian, G., Xu, C. et al. Hippo kinases MST1 and MST2 control the differentiation of the epididymal initial segment via the MEK-ERK pathway. Cell Death Differ 27, 2797–2809 (2020). https://doi.org/10.1038/s41418-020-0544-x
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DOI: https://doi.org/10.1038/s41418-020-0544-x
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