Holştayn Sığırlarında Foliküler ve Luteal Fazdaki Ovaryum Dokularında İntraovarian Genlerin Ekspresyon Profili

Abstract

Bu çalışmanın amacı, Holştayn sığırlarına ait preovülatör folikül ve korpus luteum dokularında BMP15, TGFB1, TGFB2 ve GDF9 genlerinin ekspresyon seviyelerini karşılaştırılmalı olarak belirlemektir. Bu amaç için, öncelikle dokular immunohistokimyasal boyama ile incelendi. Daha sonra foliküler sıvılar ELISA testiyle incelenerek östradiol ve progesteron seviyeleri belirlendi. Son olarak qRT-PCR ile gruplar arasında ilgili genlere ait ekspresyon seviyeleri tespit edildi. İmmunohistokimyasal boyama sonucunda preovülatör foliküllerde yoğun miktarda östrojen reseptör alfa ve progesteron reseptör immunpozitifliklerine, korpus luteum da çok hafif düzeyde östrogen alfa reseptör immunpozitifliğine, progesteron reseptörlerinin ise preovülatör foliküllerdeki pozitiflik düzeyine yakın olduğu belirlendi. Östradiol seviyesi, preovulatör foliküllerde yüksek, progesteron seviyesi ise korpus luteumda yüksek olarak bulundu. Preovülatör foliküllerdeki TGFB1 ve TGFB2 genlerine ait mRNA transkript seviyesi korpus luteuma göre istatistiksel olarak daha yüksek bulundu (p<0.01, p<0.05, sırasıyla), ancak gruplar arasında BMP15 ve GDF9 genlerine ait mRNA transkript seviyesinde istatistiksel olarak bir fark gözlenmedi (p>0.05). Sonuç olarak intraovarian genlerin farklı ekspresyonunun, ovaryum dokusunu oluşturan hücre popülasyonu içinde folikül dinamikleri ve gen ekspresyon seviyelerinin farklılıklarıyla ilişkili olabileceğini ve bu durumun da foliküler ve luteal dönemin sonucu olarak ortaya çıkabileceği düşünülmektedir. 

Keywords

• Korpus luteum,holştayn,preovulatör folikül,, intraovarian genler,qRT-PCR

Expression Profile of Intraovarian Genes in Ovary Tissues at Follicular and Luteal Phases in Holstein Cattle

Abstract

The aim of the present study is to determine comparatively expression levels of the BMP15, TGFB1, TGFB2 and GDF9 genes in the preovulatory follicle and corpus luteum tissue of Holstein cattle. For this purpose, primarily the tissues were examined by immunohistochemical staining. Later on, follicular fluid was analyzed by ELISA test and estradiol and progesterone levels were determined. Finally, expression levels of the related genes were determined between the groups by qRT-PCR. Immunohistochemical staining revealed that estrogen receptor alpha and progesterone receptor immunoreactivities were intensely present in preovulatory follicles, estrogen alpha receptor immunoreactivity was very slight and progesterone receptors were similar to positivity in preovulatory follicles in corpus luteum. Furthermore, estradiol level was high in preovulatory follicles and progesterone level was high in corpus luteum. The levels of mRNA transcripts of the TGFB1 and TGFB2 genes in the preovulatory follicles were statistically higher than the corpus luteum (p<0.01, p<0.05, respectively), but there was no statistically significant difference between the groups in the mRNA transcript levels of the BMP15 and GDF9 genes (p>0.05). As a result, it is thought that differential expression of intraovarian genes may be associated with differences in follicular dynamics and gene expression levels within the cell population of ovarian tissue, so this situation may result in follicular and luteal phase.

Keywords

• Corpus luteum,holstein,intraovarian genes,preovulatory follicle,qRT-PCR

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