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DNA integrity and methylation changes of mouse spermatozoa following prolonged incubation

Sedigheh Nematollahi1, Mehdi Mehdizadeh1, Sara Hosseini2

  • 1Department of Anatomy, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran.

Andrologia
|March 29, 2019
PubMed

Insights

Incubating mouse sperm at room temperature (RT) for 24 hours improved motility and viability, while 37°C incubation increased DNA damage and altered DNA methylation gene expression. RT incubation preserves better sperm quality.

Area of Science:

  • Reproductive Biology
  • Epigenetics
  • Spermatozoa Research

Background:

  • Sperm quality is crucial for successful fertilization and can be impacted by pre-insemination incubation times.
  • Understanding how incubation conditions affect sperm DNA methylation and integrity is vital for optimizing assisted reproductive technologies.

Purpose of the Study:

  • To investigate the effects of different incubation temperatures (room temperature vs. 37°C) on mouse spermatozoa.
  • To evaluate changes in sperm DNA methylation levels and the expression of related genes (DNMTs) following incubation.

Main Methods:

  • Spermatozoa were divided into fresh, room temperature (RT), and 37°C incubation groups for 24 hours.
  • Sperm DNA fragmentation index (DFI) was assessed using the Sperm Chromatin Structure Assay.
  • DNA methylation levels were analyzed by flow cytometry, and gene expression by quantitative real-time PCR (qRT-PCR).

Main Results:

  • Sperm incubated at RT showed significantly higher progressive motility and viability compared to 37°C incubation.
  • Incubation at 37°C resulted in a higher DFI and a significant decrease in DNA methylation levels.
  • Expression of DNA methyltransferases (DNMTs), including Dnmt-1, Dnmt-3a, Dnmt-3b, and Dnmt-3l, was altered by incubation at both RT and 37°C.

Conclusions:

  • A 24-hour incubation period significantly affects sperm DNA methylation and integrity.
  • Incubation at room temperature is preferable for maintaining better mouse sperm quality compared to 37°C incubation.
Keywords:
5-methylcytosinechromatin integrityglobal DNA methylationsperm incubation

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