Alteration of Global DNA Methylation in the Oocyte Maturation Process

Özgür Çınar

Makale detayları

Alteration of Global DNA Methylation in the Oocyte Maturation Process

Yazarlar:

Özgür Çınar

Yayıncı:

Akdeniz University

Ana Başlıklar:

Klinik Tıp Bilimleri

Dergi:

Akdeniz Tıp Dergisi Vol.10 Issue 1, pp.47-52

Belge Tipi:

Makale

Yayın Tarihi:

1/2024

Belge Dili:

Türkçe

Document ID (Org No.):

2001309833

pISSN:

2687-2781

Anahtar kelimeler:

Özet:

 Objective: In this study, it was aimed to reveal whether alteration of global DNA methylation occurs or not during the oocyte maturation from Germinal Vesicle GV to Metaphase II MII. Method: The difference in global DNA methylation between the GV and in vivo MII oocyte stages of 4-week-old BalbC mice was examined by using the immunofluorescence method. For this purpose, oocytes at GV and MII stages were labeled with anti-5-methyl cytosine 5mC antibody and images from optical sections taken with Zeiss LSM-880 Airyscan confocal microscope were obtained. Signal intensities of the images were digitalized and calculated by using Image-J software. Result: When the global DNA methylation was evaluated after 5-methyl cytosine 5mC labeling, 5mC signals were observed in nucleus of GV stage oocytes whereas, at metaphase plate of MII stage. Relative signal intensities of global DNA methylation were evaluated, and statistically significant p0.0001 3.2-fold decrease was found at MII stage compared to GV. Calculations aiming to reveal whether this significant decrease depended on the reduction in the amount of DNA after meiosis I or not, showed that the decrease in global DNA methylation may not be caused by meiosis itself. Conclusion: The result that the global DNA methylation level was decreased more than three times at the MII stage compared to the GV stage oocytes suggests that DNA methylation is controlled by various mechanisms, and this may have vital role fertilization dynamics.

 Objective: In this study, it was aimed to reveal whether alteration of global DNA methylation occurs or not during the oocyte maturation from Germinal Vesicle GV to Metaphase II MII. Method: The difference in global DNA methylation between the GV and in vivo MII oocyte stages of 4-week-old BalbC mice was examined by using the immunofluorescence method. For this purpose, oocytes at GV and MII stages were labeled with anti-5-methyl cytosine 5mC antibody and images from optical sections taken with Zeiss LSM-880 Airyscan confocal microscope were obtained. Signal intensities of the images were digitalized and calculated by using Image-J software. Result: When the global DNA methylation was evaluated after 5-methyl cytosine 5mC labeling, 5mC signals were observed in nucleus of GV stage oocytes whereas, at metaphase plate of MII stage. Relative signal intensities of global DNA methylation were evaluated, and statistically significant p0.0001 3.2-fold decrease was found at MII stage compared to GV. Calculations aiming to reveal whether this significant decrease depended on the reduction in the amount of DNA after meiosis I or not, showed that the decrease in global DNA methylation may not be caused by meiosis itself. Conclusion: The result that the global DNA methylation level was decreased more than three times at the MII stage compared to the GV stage oocytes suggests that DNA methylation is controlled by various mechanisms, and this may have vital role fertilization dynamics.

 Objective: In this study, it was aimed to reveal whether alteration of global DNA methylation occurs or not during the oocyte maturation from Germinal Vesicle GV to Metaphase II MII. Method: The difference in global DNA methylation between the GV and in vivo MII oocyte stages of 4-week-old BalbC mice was examined by using the immunofluorescence method. For this purpose, oocytes at GV and MII stages were labeled with anti-5-methyl cytosine 5mC antibody and images from optical sections taken with Zeiss LSM-880 Airyscan confocal microscope were obtained. Signal intensities of the images were digitalized and calculated by using Image-J software. Result: When the global DNA methylation was evaluated after 5-methyl cytosine 5mC labeling, 5mC signals were observed in nucleus of GV stage oocytes whereas, at metaphase plate of MII stage. Relative signal intensities of global DNA methylation were evaluated, and statistically significant p0.0001 3.2-fold decrease was found at MII stage compared to GV. Calculations aiming to reveal whether this significant decrease depended on the reduction in the amount of DNA after meiosis I or not, showed that the decrease in global DNA methylation may not be caused by meiosis itself. Conclusion: The result that the global DNA methylation level was decreased more than three times at the MII stage compared to the GV stage oocytes suggests that DNA methylation is controlled by various mechanisms, and this may have vital role fertilization dynamics.

 Amaç: Bu çalışmada, global DNA metilasyonunun Germinal Vezikül GV aşamasından Metafaz II MII aşamasına kadar olan oosit maturasyonu sürecinde değişim gösterip göstermediğinin ortaya konulması amaçlanmıştır. Yöntem: Bu çalışmada, 4 haftalık BalbC farelerinin GV ve in vivo MII oosit evreleri arasındaki global DNA metilasyonu farkı immünofloresan yöntemi kullanılarak incelendi. Bu amaçla GV ve MII aşamasındaki oositlerde 5-metil sitozin 5mC işaretlemesi sonrası Zeiss LSM-880 Airyscan konfokal mikroskopta alınan optik kesitlerinden elde edilen görüntülerden Image-J yazılımı kullanılarak hesaplanan sinyal yoğunlukları değerlendirildi. Bulgular: Global DNA metilasyonu, 5-metil sitozin 5mC işaretlemesi sonrası değerlendirildiğinde, GV aşamasındaki oositlerde, çekirdek bölgesinde gözlemlenirken, MII aşamasındaki oositlerde metafaz plağına uygun lokasyonda, olduğu izlendi. Global DNA metilasyonunun göreceli sinyal yoğunluğu değerlendirildiğinde; MII aşamasında GV aşamasına göre 3,2 katlık istatistiki olarak anlamlı bir azalma olduğu saptandı. Bu azalışın birinci mayoz sonrası DNA miktarındaki azalmaya bağlı olup olmadığı için yapılan hesaplamalar da bunun sadece DNA miktarındaki azalmadan kaynaklanmadığını gösterdi. Sonuç: Oositlerde, GV aşamasına kıyasla MII evresinde global DNA metilasyon seviyesinin üç kattan daha fazla azalmış olması, fertilizasyon öncesi oositteki DNA metilasyonunun çeşitli mekanizmalarla kontrol edildiğini ve bunun fertilizasyon dinamiğinde önemli olabileceğini göstermiştir.

Makale detayları

Alteration of Global DNA Methylation in the Oocyte Maturation Process

Yazarlar:

Özgür Çınar

Yayıncı:

Akdeniz University

Ana Başlıklar:

Klinik Tıp Bilimleri

Dergi:

Akdeniz Tıp Dergisi Vol.10 Issue 1, pp.47-52

Belge Tipi:

Makale

Yayın Tarihi:

1/2024

Belge Dili:

Türkçe

Document ID (Org No.):

2001309833

pISSN:

2687-2781

Epigenetic

Global DNA methylation

Oogenesis

OocyteEpigenetik

Global DNA metilasyonu

Oogenez

Oosit

Sayı

Makale detayları

Yazarlar: Özgür Çınar

Yayıncı: Akdeniz University

Ana Başlıklar: Klinik Tıp Bilimleri

Dergi: Akdeniz Tıp Dergisi Vol.10 Issue 1, pp.47-52

Belge Tipi: Makale

Yayın Tarihi: 1/2024

Belge Dili: Türkçe

Document ID (Org No.): 2001309833

pISSN: 2687-2781

Sayı

Yazarlar:

Özgür Çınar

Yayıncı:

Akdeniz University

Ana Başlıklar:

Klinik Tıp Bilimleri

Dergi:

Akdeniz Tıp Dergisi Vol.10 Issue 1, pp.47-52

Belge Tipi:

Makale

Yayın Tarihi:

1/2024

Belge Dili:

Türkçe

Document ID (Org No.):

2001309833

pISSN:

2687-2781