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Updated: Apr 20, 2026
A Seminiferous Tubule Squash Technique for the Cytological Analysis of Spermatogenesis Using the Mouse Model
Published on: February 6, 2018
In vivo notch signaling blockade induces abnormal spermatogenesis in the mouse
Daniel Murta1, Marta Batista1, Alexandre Trindade2
1Reproduction and Development, Interdisciplinary Centre of Research in Animal Health (CIISA), Faculty of Veterinary Medicine, University of Lisbon, Lisbon, Portugal.
Abstract:
In a previous study we identified active Notch signaling in key cellular events occurring at adult spermatogenesis. In this study, we evaluated the function of Notch signaling in spermatogenesis through the effects of in vivo Notch blockade. Adult CD1 male mice were either submitted to a long term DAPT (?-secretase inhibitor) or vehicle treatment. Treatment duration was designed to attain one half the time (25 days) or the time (43 days) required to accomplish a complete cycle of spermatogenesis. Blockade of Notch signaling was depicted from decreased transcription of Notch effector genes. Notch signaling blockade disrupted the expression patterns of Notch components in the testis, induced male germ cell fate aberrations, and significantly increased germ cell apoptosis, mainly in the last stages of the spermatogenic cycle, and epididymis spermatozoa morphological defects. These effects were more pronounced following the 43 day than the 25 day DAPT treatment schedule. These results indicate a relevant regulatory role of Notch signaling in mammalian spermatogenesis.
Insights
Notch signaling is crucial for male fertility. Blocking this pathway in mice disrupted sperm development, causing germ cell abnormalities and reduced sperm quality, highlighting its regulatory role in spermatogenesis.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Molecular Endocrinology
Background:
- Notch signaling is active in adult spermatogenesis.
- Its precise function in male germ cell development requires further elucidation.
Purpose of the Study:
- To investigate the functional role of Notch signaling in mammalian spermatogenesis.
- To evaluate the consequences of in vivo Notch pathway blockade on sperm development.
Main Methods:
- Adult male mice received long-term treatment with DAPT (a ?-secretase inhibitor) or a vehicle control.
- Treatment durations were 25 days or 43 days, corresponding to half or a full cycle of spermatogenesis.
- Changes in Notch effector gene transcription, gene expression, germ cell apoptosis, and sperm morphology were assessed.
Main Results:
- Notch signaling blockade led to decreased transcription of Notch effector genes.
- Disrupted expression of Notch components in the testis.
- Induced male germ cell fate aberrations and increased germ cell apoptosis, particularly in later stages.
- Significant morphological defects in epididymis spermatozoa were observed.
- Effects were more severe with longer DAPT treatment (43 days vs. 25 days).
Conclusions:
- Notch signaling plays a significant regulatory role in mammalian spermatogenesis.
- Disruption of Notch signaling impairs germ cell development and sperm quality.
- The findings underscore the importance of the Notch pathway for male reproductive health.
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