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Updated: May 19, 2026
Isolation of Murine Spermatogenic Cells using a Violet-Excited Cell-Permeable DNA Binding Dye
Published on: January 14, 2021
Proteomic changes in rat spermatogenesis in response to in vivo androgen manipulation; impact on meiotic cells
Peter G Stanton1, Pavel Sluka, Caroline F H Foo
1Prince Henry's Institute, Monash Medical Centre, Clayton, Victoria, Australia. peter.stanton@princehenrys.org
Abstract:
The production of mature sperm is reliant on androgen action within the testis, and it is well established that androgens act on receptors within the somatic Sertoli cells to stimulate male germ cell development. Mice lacking Sertoli cell androgen receptors (AR) show late meiotic germ cell arrest, suggesting Sertoli cells transduce the androgenic stimulus co-ordinating this essential step in spermatogenesis. This study aimed to identify germ cell proteins responsive to changes in testicular androgen levels and thereby elucidate mechanisms by which androgens regulate meiosis. Testicular androgen levels were suppressed for 9 weeks using testosterone and estradiol-filled silastic implants, followed by a short period of either further androgen suppression (via an AR antagonist) or the restoration of intratesticular testosterone levels. Comparative proteomics were performed on protein extracts from enriched meiotic cell preparations from adult rats undergoing androgen deprivation and replacement in vivo. Loss of androgenic stimulus caused changes in proteins with known roles in meiosis (including Nasp and Hsp70-2), apoptosis (including Diablo), cell signalling (including 14-3-3 isoforms), oxidative stress, DNA repair, and RNA processing. Immunostaining for oxidised DNA adducts confirmed spermatocytes undergo oxidative stress-induced DNA damage during androgen suppression. An increase in PCNA and an associated ubiquitin-conjugating enzyme (Ubc13) suggested a role for PCNA-mediated regulation of DNA repair pathways in spermatocytes. Changes in cytoplasmic SUMO1 localisation in spermatocytes were paralleled by changes in the levels of free SUMO1 and of a subunit of its activating complex, suggesting sumoylation in spermatocytes is modified by androgen action on Sertoli cells. We conclude that Sertoli cells, in response to androgens, modulate protein translation and post-translational events in spermatocytes that impact on their metabolism, survival, and completion of meiosis.
Insights
Androgens regulate sperm production by acting on Sertoli cells, which influences germ cell proteins involved in meiosis, DNA repair, and survival. This study reveals how androgens control key pathways for successful spermatogenesis.
Area of Science:
- Reproductive Biology
- Molecular Endocrinology
- Spermatogenesis
Background:
- Sperm production requires androgen signaling, primarily through Sertoli cells, to regulate germ cell development and meiosis.
- Sertoli cell androgen receptors (AR) are crucial for transducing androgenic signals that coordinate spermatogenesis.
- Understanding how androgens impact germ cell proteins is key to elucidating mechanisms of meiotic regulation.
Purpose of the Study:
- To identify germ cell proteins affected by testicular androgen levels.
- To elucidate the mechanisms by which androgens regulate meiosis in spermatogenesis.
- To investigate the role of Sertoli cell-mediated androgen action on spermatocyte proteins.
Main Methods:
- Adult rats underwent 9 weeks of androgen suppression followed by androgen suppression or restoration.
- Comparative proteomics analyzed protein extracts from enriched meiotic cell preparations.
- Immunostaining assessed oxidative DNA damage, and protein levels of PCNA, Ubc13, and SUMO1 were evaluated.
Main Results:
- Androgen deprivation altered proteins involved in meiosis, apoptosis, cell signaling, oxidative stress, DNA repair, and RNA processing.
- Spermatocytes showed oxidative stress-induced DNA damage and increased PCNA and Ubc13 during androgen suppression.
- Changes in SUMO1 localization and levels indicated modified sumoylation in spermatocytes due to androgen action.
Conclusions:
- Sertoli cells, under androgen influence, modulate protein translation and post-translational modifications in spermatocytes.
- These modulations impact spermatocyte metabolism, survival, and the successful completion of meiosis.
- Androgen signaling via Sertoli cells is critical for maintaining germ cell protein homeostasis and meiotic progression.
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