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  4. CHARACTERIZING BRDT FUNCTION IN MAMMALIAN SPERMATOGENESIS: GENETIC MODELS AND THERAPEUTIC IMPLICATIONS FOR NON-HORMONAL CONTRACEPTION

CHARACTERIZING BRDT FUNCTION IN MAMMALIAN SPERMATOGENESIS: GENETIC MODELS AND THERAPEUTIC IMPLICATIONS FOR NON-HORMONAL CONTRACEPTION

File(s)
Simon_cornellgrad_0058F_15076.pdf (31.04 MB)
No Access Until
2026-09-09
Permanent Link(s)
https://doi.org/10.7298/sfyk-4m16
https://hdl.handle.net/1813/120775
Collections
Cornell Theses and Dissertations
Author
Simon, Leah
Abstract

The testis-specific bromodomain and extra-terminal domain (BET) protein BRDT is a key regulator of spermatogenesis and a compelling target for non-hormonal male contraceptive development. Despite significant interest in the regulatory function and contraceptive potential of BRDT, fundamental questions remain about how BRDT governs critical processes during meiosis and spermiogenesis. These gaps largely stem from the historical absence of genetic models and molecular tools necessary to dissect the stage-specific roles of BRDT in the germline. A primary focus of this dissertation is to explore the effects of transient BRDT inhibition using the small-molecule inhibitor (+)-JQ1. Single-cell transcriptomic and cytological analyses reveal that BRDT inhibition leads to post-meiotic germ cell loss, followed by full recovery upon drug withdrawal. These findings demonstrate that meiosis is a reversible point of intervention and that BRDT disruption selectively affects direct transcriptional targets, highlighting its essential role in orchestrating meiotic gene expression and chromosomal events during prophase I. To further characterize BRDT function, this dissertation introduces two novel mouse models – a knockout line and an epitope-tagged line. The Brdt-/- line exhibits dual-stage defects in meiosis and spermiogenesis, underscoring the stage-specific regulatory roles of BRDT. In parallel, the BRDT epitope-tagged line enables reliable detection of the endogenous protein, overcoming longstanding technical barriers due to limited antibody availability. To address the broad regulatory influence of BRDT, this work identifies a refined list of germline-restricted, testis-specific target genes with greater potential for selective contraceptive intervention. It also establishes proof-of-concept for gene manipulation in enriched spermatocytes and spermatids via lentiviral transduction, enabling future CRISPR-based screens. Finally, novel in vitro platforms for meiotic initiation and progression provide a foundational framework for functional studies in defined germ cell stages. Collectively, this dissertation positions BRDT as a central coordinator of male germ cell development and a gateway to identifying downstream targets with high contraceptive potential. Through genetic modeling, transcriptomic analysis, and foundational work in germ cell manipulation, this research offers new tools and insights to support the development of safe, reversible, and testis-specific strategies for male contraception.

Description
265 pages
Date Issued
2025-08
Keywords
BRDT
•
Genetics
•
Mouse
•
Spermatogenesis
Committee Chair
Cohen, Paula
Committee Member
Travis, Alexander
Wolfner, Mariana
Roberson, Mark
Degree Discipline
Biomedical and Biological Sciences
Degree Name
Ph. D., Biomedical and Biological Sciences
Degree Level
Doctor of Philosophy
Type
dissertation or thesis

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