Expression pattern of notch2 in the fetal lung tissue of mice obtained by in vitro embryo culture and embryo transfer

Yıl 2025, Cilt: 14 Sayı: 2, 25 - 30, 25.12.2025

Göksel Doğan , Levent Karagenç

https://doi.org/10.53913/aduveterinary.1746913

Öz

Notch is a critical regulator of differentiation of proximal airway epithelial cells and development of alveoli. Any defect in Notch2 signaling (pathway) impairs alveolar development in mice. The purpose of the current study was to determine the levels of mRNA encoding Notch2 and to reveal the expression pattern of Notch2 in the fetal lung tissues of mice obtained by the transfer of blastocyst to pseudopregnant females. Two groups (control group: CG, and experimental group: EG) were included in the study. Experimental fetuses were produced by transfer of in vitro cultured blastocysts. Mice obtained after natural mating served as Control group. The transcriptional expression of Notch2 was determined with quantitative-Real time PCR (qRT-PCR). Expression pattern of Notch2 was detected using immunohistochemistry. Results of qRT-PCR indicated that the levels of mRNA encoding Notch2 were similar between the groups. Results of immunohistochemistry demontrated that Notch2 immunopositivity was observed in lung tissues of both CG and EG fetuses. Results further demonstrated that Notch2 was strongly expressed in the bronchi, bronchioles, epithelial cells of terminal sacculus, vascular endothelial cells and interstitial area in the lung tissues of CG fetuses. Although the expression was weak, Notch2 was detected in the glandular structures that will form the alveoli, as well as in the bronchi, bronchioles, vascular endothelial cells and interstitial area in the lung tissues of EG fetuses. In any case, details surrounding if and to what extent Notch2 plays a role in the regulation of lung development as a consequence of in vitro embryo culture and embryo transfer warrants further investigations.

Anahtar Kelimeler

embryo transfer , in vitro embryo culture , lung development , notch2 , mouse

Etik Beyan

All animal experiments were certificated by the institutional Animal Ethics Committee of Adnan Menderes University, Aydin, Turkey (protocol no: 64583101/2014/151).

Destekleyen Kurum

The present study was conducted without any financial support.

İn vitro embriyo kültürü ve embriyo transferi ile elde edilen farelerde fetal akciğer dokusunda Notch2 ekspresyon paterni

Öz

Notch solunum yolu epitel hücrelerinin farklılaşmasında ve alveollerin gelişiminde kritik bir düzenleyicidir. Notch2 sinyal yolağında gözlenen herhangi bir defekt farelerde alveolar gelişimi bozmaktadır. Bu çalışmanın amacı, in vitro kültürü yapılan blastosistlerin yalancı gebe dişilere transferiyle oluşturulan farelere ait fetal akciğer dokularında Notch2 ekspresyonunun değişip değişmediğini analiz etmek ve bu farelerin akciğer dokularında Notch2 ekspresyon paternini belirlemektir. Çalışmada bir deneme ve bir kontrol grubu bulunmaktadır. Deneme grubuna ait fetüsler (EG), in vitro kültürü yapılan blastosistlerin yalancı gebe dişilere transferi yoluyla elde edilmişlerdir. Doğal olarak yumurtlayan dişilerden elde edilen fareler ise kontrol grubunu (CG) oluşturmuştur. Notch2' nin transkripsiyonel ekspresyonunu belirlemek amacıyla kantitatif-gerçek zamanlı PCR (qRT-PCR) kullanılmıştır. Notch2 ekspresyon paterni immünohistokimya yöntemiyle belirlenmiştir. Elde edilen veriler, Notch2 'nin transkripsiyonel ekspresyonunun her iki grupta da benzer olduğunu göstermektedir. Hem kontrol ve hem de deneme gruplarına ait fetal akciğer dokularında Notch2 immün-pozitifliği gözlemlenmiştir. Notch2 kontrol grubuna ait akciğer dokularında (CG) bronş, bronşiyol, endotelyal hücreler ve sakkuler keseleri çevreleyen epitel hücrelerinde baskın şekilde eksprese edilmektedir. Deneme grubu fetüslerinde (EG) ise Notch2 ekspresyonu hafif bir şekilde gözlemlenmiştir. Notch2 bronş, bronşiyol, endotelyal hücreler ve intersitisyel alanlarda görülmekle birlikte ileride alveolleri oluşturacak olan bezsel yapılarda da eksprese edilmektedir. Ancak, Notch2' nin in vitro embriyo kültürü ve embriyo transferine bağlı olarak akciğer gelişiminin düzenlenmesinde bir rol oynayıp oynamadığı veya ne ölçüde rol oynadığı hala belirsizliğini korumaktadır ve bu nedenle daha fazla araştırma yapılması gerekmektedir.

Anahtar Kelimeler

in vitro embriyo kültürü , embriyo transferi , akciğer gelişimi , notch2 , fare

Etik Beyan

Tüm hayvan deneyleri Adnan Menderes Üniversitesi, Hayvan Deneyleri Etik Kurulu tarafından onaylanmıştır (protokol no: 64583101/2014/151).

Destekleyen Kurum

Sunulan çalışma herhangi bir finansal destek almadan gerçekleştirilmiştir.

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