Thesis
Chromatin Dynamics During Murine Spermatogonial Development
Washington State University
Master of Science (MS), Washington State University
01/2022
DOI:
https://doi.org/10.7273/000004589
Handle:
https://hdl.handle.net/2376/124774
Abstract
Male infertility is a growing concern in our world today. Issues with spermatogenesis, specifically spermatogonial development, can lead to infertility or subfertility in men. Within the complex process of spermatogenesis, the germ cells have profound transcriptional and morphological changes. One key point in spermatogenesis where the aforementioned changes occur is at the point of spermatogonial differentiation. This differentiation event is mediated by retinoic acid (RA) to irreversibly commit the germ cells to spermatogenesis, inciting transcriptional and morphological changes. While these two changes have received great attention, there is a lack of understanding of epigenetic mechanisms influencing male germ cell development. Previous studies using transmission electron microscopy have revealed that chromatin compaction increases during spermatogonial development; however, it is unknown where in the genome this compaction occurs. Within this work, we aimed to better understand the dynamics of chromatin accessibility in relation to RA action during spermatogonial development via ATAC-seq on multiple spermatogonial subtypes. By synchronizing the testis, we were able to isolate four subpopulations of spermatogonia corresponding with undifferentiated spermatogonia, A1/A2 spermatogonia, A3 spermatogonia, and B spermatogonia. These ATAC-seq data revealed that following RA administration, there was over a 59% decrease of accessible regions within the genome. Of all assayed cell populations, 68-89% of all accessible regions were within 10 kb of an annotated gene, and 72.8% remained accessible across all four time points. Further analysis showed the majority of these accessible genes were accessible at, or just upstream of their transcription start site (TSS) at all time points. Investigation of spermatogonial genes (Neurog3, Sox3, Zbtb16, Lin28a, Nanos2, Pou5f1), as well as common immunohistochemical markers of differentiating spermatogonia (Kit, Stra8), revealed accessible chromatin at all assayed timepoints largely surrounding the TSS. Together these data support the conclusion that RA is triggering large-scale chromatin condensation. However, it appears this condensation of the chromatin is more structural rather than transcriptional in nature.
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Details
- Title
- Chromatin Dynamics During Murine Spermatogonial Development
- Creators
- Mary Christine Schleif
- Contributors
- Michael D Griswold (Advisor)Patricia A Hunt (Committee Member)Wipawee Winuthayanon (Committee Member)Jon M Oatley (Committee Member)
- Awarding Institution
- Washington State University
- Academic Unit
- Molecular Biosciences, School of
- Theses and Dissertations
- Master of Science (MS), Washington State University
- Publisher
- Washington State University
- Number of pages
- 66
- Identifiers
- OCLC#: 1371068737; 99900898938901842
- Language
- English
- Resource Type
- Thesis