DAZL mediates a broad translational program regulating expansion and differentiation of spermatogonial progenitors
- Whitehead Institute, United States
Abstract
Fertility across metazoa requires the germline-specific DAZ family of RNA-binding proteins. Here we examine whether DAZL directly regulates progenitor spermatogonia using a conditional genetic mouse model and in vivo biochemical approaches combined with chemical synchronization of spermatogenesis. We find that the absence of Dazl impairs both expansion and differentiation of the spermatogonial progenitor population. In undifferentiated spermatogonia, DAZL binds the 3' UTRs of ~2,500 protein-coding genes. Some targets are known regulators of spermatogonial proliferation and differentiation while others are broadly expressed, dosage-sensitive factors that control transcription and RNA metabolism. DAZL binds 3' UTR sites conserved across vertebrates at a UGUU(U/A) motif. By assessing ribosome occupancy in undifferentiated spermatogonia, we find that DAZL increases translation of its targets. In total, DAZL orchestrates a broad translational program that amplifies protein levels of key spermatogonial and gene regulatory factors to promote the expansion and differentiation of progenitor spermatogonia.
Data availability
All sequencing data generated in this study are available at NCBI Gene Expression Omnibus accession number GSE145177
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Gene expression in Pou5f1:EGFP-positive spermatogonia from Dazl conditional knockout and control malesNCBI Gene Expression Omnibus (GEO), GSE144919.
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Gene expression in undifferentiated spermatogoniaNCBI Gene Expression Omnibus (GEO), GSE144923.
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DAZL targets in undifferentiated spermatogoniaNCBI Gene Expression Omnibus (GEO), GSE144920.
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Translational profiling in undifferentiated spermatogoniaNCBI Gene Expression Omnibus (GEO), GSE144922.
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DAZL mediates a broad translational program regulating expansion and differentiation of spermatogonial progenitorsNCBI Gene Expression Omnibus (GEO), GSE145177.
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RNA-seq in spermatogonia from PRC1ctrl and dKO miceNCBI Gene Expression Omnibus (GEO), GSE102783.
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Analysis of gene expression in populations of adult undifferentiated spermatogoniaNCBI Gene Expression Omnibus (GEO), GSE107124.
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Dazl maintains proliferating germ cells through a network of polyA-proximal mRNA interactions [P6 iCLIP]NCBI Gene Expression Omnibus (GEO), GSE108183.
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Conservation, acquisition, and functional impact of sex-biased gene expression in mammalian tissuesNCBI Gene Expression Omnibus (GEO), GSE125483.
Article and author information
Author details
Funding
Howard Hughes Medical Institute (Page laboratory)
- David C Page
Lalor Foundation (Postdoctoral fellowship)
- Maria M Mikedis
Eunice Kennedy Shriver National Institute of Child Health and Human Development (F32HD093391)
- Maria M Mikedis
National Natural Science Foundation of China (81471507)
- Yuting Fan
National Key Research and Development Program of China (2017YFC1001600)
- Yuting Fan
Hope Funds for Cancer Research (HFCR-15-06-06)
- Peter K Nicholls
National Health and Medical Research Council (GNT1053776)
- Peter K Nicholls
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: All experiments involving mice were performed in accordance with the guidelines of the Massachusetts Institute of Technology (MIT) Division of Comparative Medicine, which is overseen by MIT's Institutional Animal Care and Use Committee (IACUC). The animal care program at MIT/Whitehead Institute is accredited by the Association for Assessment and Accreditation of Laboratory Animal Care, International (AAALAC), and meets or exceeds the standards of AAALAC as detailed in the Guide for the Care and Use of Laboratory Animals. The MIT IACUC approved this research (no. 0617-059-20).
Copyright
© 2020, Mikedis et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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DAZL mediates a broad translational program regulating expansion and differentiation of spermatogonial progenitors
eLife 9:e56523.
https://doi.org/10.7554/eLife.56523
Further reading
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- Developmental Biology
Although the impact of gender-affirming hormone therapy (GAHT) on spermatogenesis in trans women has already been studied, data on its precise effects on the testicular environment is poor. Therefore, this study aimed to characterize, through histological and transcriptomic analysis, the spermatogonial stem cell niche of 106 trans women who underwent standardized GAHT, comprising estrogens and cyproterone acetate. A partial dedifferentiation of Sertoli cells was observed, marked by the co-expression of androgen receptor and anti-Müllerian hormone which mirrors the situation in peripubertal boys. The Leydig cells also exhibited a distribution analogous to peripubertal tissue, accompanied by a reduced insulin-like factor 3 expression. Although most peritubular myoid cells expressed alpha-smooth muscle actin 2, the expression pattern was disturbed. Besides this, fibrosis was particularly evident in the tubular wall and the lumen was collapsing in most participants. A spermatogenic arrest was also observed in all participants. The transcriptomic profile of transgender tissue confirmed a loss of mature characteristics - a partial rejuvenation - of the spermatogonial stem cell niche and, in addition, detected inflammation processes occurring in the samples. The present study shows that GAHT changes the spermatogonial stem cell niche by partially rejuvenating the somatic cells and inducing fibrotic processes. These findings are important to further understand how estrogens and testosterone suppression affect the testis environment, and in the case of orchidectomized testes as medical waste material, their potential use in research.
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- Developmental Biology
- Stem Cells and Regenerative Medicine
Niches are often found in specific positions in tissues relative to the stem cells they support. Consistency of niche position suggests that placement is important for niche function. However, the complexity of most niches has precluded a thorough understanding of how their proper placement is established. To address this, we investigated the formation of a genetically tractable niche, the Drosophila Posterior Signaling Center (PSC), the assembly of which had not been previously explored. This niche controls hematopoietic progenitors of the lymph gland (LG). PSC cells were previously shown to be specified laterally in the embryo, but ultimately reside dorsally, at the LG posterior. Here, using live-imaging, we show that PSC cells migrate as a tight collective and associate with multiple tissues during their trajectory to the LG posterior. We find that Slit emanating from two extrinsic sources, visceral mesoderm and cardioblasts, is required for the PSC to remain a collective, and for its attachment to cardioblasts during migration. Without proper Slit-Robo signaling, PSC cells disperse, form aberrant contacts, and ultimately fail to reach their stereotypical position near progenitors. Our work characterizes a novel example of niche formation and identifies an extrinsic signaling relay that controls precise niche positioning.
