In vitro embriyo kültürü ve embriyo transferi ile elde edilen erişkin farelerin akciğer dokusunda p63 izoformlarının ekspresyonu

Öz

Bazal hücre biyolojisinde önemli bir rol oynayan p63 geni, TAp63 ve ΔNp63 olmak üzere iki p63 izoformunu kodlar. Daha önceki çalışmalarımızdan elde ettiğimiz veriler, kontrol grubu ile kıyaslandığında in-vitro embriyo kültürü ve embriyo transferi ile elde edilen fare fetüslerine ait akciğer dokularında TAp63 ve ΔNp63 izoformlarının mRNA ekspreyon düzeylerinin önemli ölçüde arttığını göstermektedir. Ancak, in-vitro embriyo kültürü ve transferi ile elde edilen fetüslerin akciğer dokularındaki TAp63 ve ΔNp63 izoformlarının değişen bu ifade düzeylerinin yetişkin dönemde devam edip etmediği bilinmemektedir. Sunulan bu çalışma, atmosferik oksijen koşullarında büyütülen blastosistlerin yalancı gebe dişilere transferi ile elde edilen yetişkin farelerin akciğer dokularında TAp63 ve ΔNp63 izoformlarına ait mRNA düzeylerinde herhangi bir değişiklik olup olmadığını araştırmayı amaçlamaktadır. Çalışmada bir Kontrol (CG) ve bir Deney (EG) grubu yer almaktadır. TAp63 ve ΔNp63'ün akciğer dokularındaki ekspresyon düzeylerini değerlendirmek amacıyla kantitatif real time-PCR (qRT-PCR) yöntemi kullanılmıştır. Elde edilen veriler, iki grup arasında TAp63 ve ΔNp63 mRNA ekspresyon düzeylerinde istatistiksel olarak anlamlı bir fark olmadığını göstermiştir. Ancak yetişkin dönemde TAp63 ve ΔNp63 ekspresyonları transkripsiyonel düzeylerde gruplar arasında dengelenmiş gibi gözükse de p63 izoformlarının yetişkin akciğer dokusunda translasyonel düzeylerde değişip değişmediği bilinmemektedir. Dolayısıyla, p63 izoformlarının in-vitro embriyo kültürü ve embriyo transferi ile elde edilen yetişkin fare akciğer dokularında hücre proliferasyonu, hücre farklılaşması ve apoptoz gibi hücresel mekanizmalarda rol oynayıp oynamadığını değerlendirmek amacıyla TUNEL, immünohistokimya, immünofloresan, western blott ve ELISA gibi yöntemler kullanılarak daha ileri araştırmalara ihtiyaç vardır. Ayrıca, TAp63 ve ΔNp63 ekspresyonu gösteren bazal hücre benzeri akciğer hücrelerinin, in-vitro embriyo kültürü ve embriyo transferine bağlı olarak hücre proliferasyonu, hücre farklılaşması ve apoptozdaki potansiyel rollerini açıklığa kavuşturmak için fonksiyonel çalışmalara ihtiyaç vardır.

Anahtar Kelimeler

• Embriyo transferi
• fare akciğeri
• in-vitro embriyo kültürü
• p63 izoformları
• TAp63 ve ΔNp63

Pulmonary expression of p63 isoforms in adult mice generated by in vitro embryo culture and embryo transfer

Öz

The p63 gene, which plays a crucial role in basal cell biology, encodes two primary isoforms, TAp63 and ΔNp63. Evidence indicated that the TAp63 and ΔNp63 isoforms were significantly upregulated in the lung tissues of fetuses obtained from in vitro embryo culture and embryo transfer, compared to control fetuses derived from naturally ovulating females. However, it remains unclear whether the altered expression of TAp63 and ΔNp63 isoforms observed in the lung tissues of fetuses obtained by in vitro embryo culture and transfer persists into adulthood. The current study aimed to investigate the transcriptional expression levels of TAp63 and ΔNp63 in the lung tissues of adult mice derived from transfer of blastocyst cultured under the atmospheric oxygen conditions. The study included one Control (CG) and one Experimental (EG) group. The transcript levels of TAp63 and ΔNp63 in lung tissues was determined by quantitative real-time- PCR (qRT-PCR). Data of present study indicated that the transcriptional expression levels of TAp63 and ΔNp63 were similar between two groups. However, although it seems to be balanced the transcriptional levels of TAp63 and ΔNp63 between groups, it remains unclear whether the translational levels of TAp63 and ΔNp63 alter in adult lung. To assess whether p63 isoforms have possible roles in cellular mechanisms including proliferation, differentiation and apoptosis in the lung tissues of adult mice derived from in vitro embryo culture and embryo transfer, further invastigation are warrant using multiple methods such as TUNEL, immunohistochemistry, immunofluorescence, western blotting and ELISA. Functional studies are also needed to clarify the potential roles of basal cell-like pulmonary cells expressing TAp63 and ΔNp63 in cell proliferation, cell differentiation and apoptosis as a consequence of in vitro embryo culture and embryo transfer.

Anahtar Kelimeler

• embryo transfer
• in vitro embryo culture
• mouse lung
• p63 isoforms
• TAp63 and ΔNp63

Kaynakça

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