Mehmet Osman ATLI, SEYİT ALİ KAYIŞ, Aydın GÜZELOĞLU, Selim ASLAN, Ahmet SEMACAN, Ercan KURAR, SEFA ÇELİK

Expression of epidermal growth factor (EGF) and heparin-binding EGF (HB-EGF) mrna in mare endometrium

EGF ve HB-EGF implantasyon sırasında ve embriyonik gelişim esnasında çok önemli roller oynamaktadır. Bu çalışmanın amacı, erken gebelik ve östrus siklusunda at endometriumunda EGF ve HB-EGF ekspresyon profilini karakterize etmektir. Endometrium biyopsileri östrustaki kısraklardan ovulasyon gününde (d0), geç diöstrusta (LD) ve luteolizis sonrası (AL) dönemlerde toplandı. Gebe kısraklardan ise gebeliğin 14. (P14), 18. (P18), 22. (P22) ve 60. (P60) günlerinde biyopsiler toplandı. Nispi mRNA ekspresyon seviyeleri real-time PZR kullanılarak tespit edildi. Normalize edilmiş veriler karışık model kullanılarak analiz edildi ve istatistiki olarak farklı olan gruplar Asgari Önemli Fark (AÖF) testi ile farklı gruplar tespit edildi. EGF mRNA ekspresyonu LD’de d0 göre artmasına rağmen, HB-EGF ekspresyonu siklus boyunca değişmedi. EGF ayrıca erken gebelik boyunca arttı ve en yüksek seviyeye 60. günde ulaştı. Benzer şekilde HB-EGF mRNA seviyesi 60. günde arttı. Gebelik, EGF ekspresyonunu 14. ve 18. günlerde LD ve AL göre arttırırken, HB-EGF sadece gebeliğin 18. gününde AL’e göre önemli oranda arttı. Bu sonuçlar, EGF’nin P4 seviyesinin yüksek olduğu geç diöstrusta ve gebeliğe bağlı olarak arttığını göstermektedir. HB-EGF ise gebeliğin daha ileri dönemlerinde uyarılmaktadır. Sonuç olarak, kısrak endometriumunda EGF ve HB-EGF’nin peri-implantasyon dönemindeki görülen olaylara katıldığı gözlenmiştir.

Anahtar Kelimeler:

at, biyopsi, diöstrüs, embriyo içeyerleşimi, embriyonik gelişim, embriyo, endometriyum, epidermal büyüme faktörü, gen ifadesi, genler, heparin, luteolizis, kısrak, elçi RNA, kızgınlık dönemi, kızgınlık, ovulasyon, gebelik, ters transkriptaz

Kısrak endometriumunda epidermal growth factor (EGF) ve heparin-binding EGF (HB-EGF) mrna ekspresyonu

EGF and HB-EGF play crucial roles in embryonic development and peri-implantation. In this study, aim was to characterize expression profiles of EGF and HB-EGF in mare endometrium during estrous cycle and early pregnancy. Endometrium biopsies were obtained from mares on day of ovulation (d0), late diestrus (LD) and after luteolysis (AL) in the estrus phase. In pregnant groups, biopsies were taken on days 14 (P14), 18 (P18), 22 (P22) and 60 (P60). Relative expression levels of genes were quantified using real-time RT-PCR. A mixed model was fitted on the normalized data and Least Significant Difference (LSD) test was employed to determine significantly different group(s). EGF mRNA expression was up-regulated at LD compared to d0 while HB-EGF expression was not changed throughout the cycle. EGF expression was also increased during early pregnancy with the highest expression level observed on P60. Similarly, HB-EGF mRNA level was increased on P60. Pregnancy induced EGF expression on P14 and P18 compared to LD and AL whereas expression of HB-EGF was only significantly higher on P18 than that of AL. These results indicate that EGF expression is up-regulated during the cycle at late diestrus when P4 is high and is increased by pregnancy. HB-EGF expression is induced later in the pregnancy. In conclusion, EGF and HB-EGF appear to involve in the events that happen in the mare endometrium during peri-implantation period.

Keywords:

horses, biopsy, dioestrus, embryo implantation, embryonic development, embryos, endometrium, epidermal growth factor, gene expression, genes, heparin, luteolysis, mares, messenger RNA, oestrous cycle, oestrus, ovulation, pregnancy, reverse transcriptase,

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