Farklı nüklear matürasyon aşamalarında oosit, kumulus hücreleri ve follikül sıvısında eksprese edilen matürasyon faktörlerinin rolü
Loading...
Files
Date
2016-08-05
Authors
Kuşpınar, Göktan
Journal Title
Journal ISSN
Volume Title
Publisher
Uludağ Üniversitesi
Abstract
İn vitro fertilizasyon uygulamalarında kontrollü ovaryan hiperstimülasyon sonrası yaklaşık % 8.6-15.2 oranında en az bir adet mayoz bölünmesini tamamlamamış oosit gelişimi gözlemlenir. Mayotik gelişim kilit düzenleyici proteinlerin kontrolü altındadır ve herhangi bir değişim matürasyon yetmezliği ile sonuçlanır. Kisspeptin (Kiss1) KISS1 geninin ürünü olan peptittir. Merkezi sinir sistemi düzeyinde ovaryan fonksiyonları düzenlemesindeki rolleri tanınmış olsa da, ovulasyonun kontrolündeki lokal rolleri tam olarak tanımlanamamıştır. İlk veriler hayvan çalışmalarında final oosit matürasyonundaki rolünü göstermiştir. Follikül rüptüründe ve kumulus ekspansiyonunda siklooksijenaz-2 (Cox-2) enzimatik olarak önemli rol oynamaktadır. Ovulasyonu ve follikül atılımını engellediği bilinen Cox-2 inhibitörünün, sıçan ovaryumunda seçici şekilde KISS1 ekspresyonunu baskıladığı gösterilmiştir. Bu veriler Kisspeptin ekspresyonunun Cox-2 ekspresyonu ile bağlantılı olabileceğini düşündürmektedir. Bu çalışmada matürasyonunu tamamlayamamış oositlerde ve kumulus hücrelerinde Kisspeptin ve reseptörü (Kiss1R) ile Cox-2 ekspresyon düzeyinin ve folliküler sıvı ile serum düzeyindeki konsantrasyonlarının oosit matürasyonundaki rollerini değerlendirmek hedeflendi. Kontrollü ovaryan stimülasyon döngülerinden elde edilen immatür ve matür oositler, follikül sıvıları ve kumulus hücreleri, hasta serum örnekleri çalışma kapsamında incelendi. Serum ve follikül sıvılarında ELİZA yöntemiyle, oosit ve kumulus hücrelerinde gerçek zamanlı polimeraz zincir reaksiyonu ile Kiss1 ve Kiss1R ve Cox-2 seviyelerine bakıldı. Elde edilen sonuçların birbirleriyle ve hastaların klinik verileriyle korelasyonları değerlendirildi. Serum ve folliküler sıvı düzeyinde Kiss1, Kiss1R ve Cox-2 konsantrasyonlarının oositin matürasyon seviyesi ile korele olmadığı görüldü. İmmatür oositlerde ve kumulus hücrelerinde Kiss1 ve reseptöründe azalma görülürken, Cox-2 ekspresyonunda artış gözlendi. Kontrollü ovaryan stimülasyon döngülerinde Kiss1 ve reseptörünün oosit matürasyon sürecinde mikroçevre düzeyinde etkin olduğu, Cox-2'nin negatif etkisinin ovulasyon öncesi final matürasyon basamağında kontrol edici rolünün olabileceği sonucuna varıldı.
In in vitro fertilization treatment following contolled ovarian hyperstimulation protocols, approximately 8.6% to 15.2% of all infertility patients at least produce one meiotically incompetent oocyte. Meiotic development is under the control of critical regulatory proteins and any alterations in this process results in meiotic maturation defects. Kisspeptin (Kiss1) is a peptide expressed by the KISS1 gene. Its role in the regulation of ovarian function at the level of central nervous system is known, however its local roles in the control of ovulation have not exactly been identified. Initial data from animal studies demonstrated the role of Kisspeptin in the final maturation of the oocyte. Cyclooxygenase-2 (Cox-2) has an important enzymatic role in cumulus expansion and follicle rupture. Cox-2 inhibitors, known as an inhibitor of ovulation and follicle rupture, has been shown to selectively suppress KISS1 expression in rat ovaries. All these data give rise to the thought that kisspeptin expression may be associated with Cox-2 expression. In this study, it is aimed to reveal the role of Kisspeptin, Kisspeptin receptor (Kiss1R) and Cox-2, on the oocyte maturation by evaluating the expression levels of Kiss1, Kiss1R and Cox-2 in immature oocytes and cumulus cells, as well as their concentration levels in serum and follicle fluids. In this study, immature and mature oocytes, cumulus cells and follicular fluids obtained from controlled ovarian stimulation cycles and serum from patients were examined. Kiss1, Kiss1R and Cox-2 levels were evaluated in oocytes and cumulus cells by real-time polymerase chain reaction, and in serum and follicle fluid by ELISA. The results were evaluated as the correlation to each other and to the clinical data of the patients. The levels of Kiss1, KissR1 and Cox-2 concentration were shown to not correlate with oocytes maturation level in serum and follicular fluid. While a decrease in the expression of Kiss1 and its receptor was seen in immature oocytes and cumulus cells, an increase in Cox-2 expression was observed. The results of this study suggested that, in controlled ovarian stimulation cycles, Kiss1 and its receptors are effective in oocytes maturation process at the level of microenvironment, and that the negative effects of Cox-2 may control the final maturation step before ovulation.
In in vitro fertilization treatment following contolled ovarian hyperstimulation protocols, approximately 8.6% to 15.2% of all infertility patients at least produce one meiotically incompetent oocyte. Meiotic development is under the control of critical regulatory proteins and any alterations in this process results in meiotic maturation defects. Kisspeptin (Kiss1) is a peptide expressed by the KISS1 gene. Its role in the regulation of ovarian function at the level of central nervous system is known, however its local roles in the control of ovulation have not exactly been identified. Initial data from animal studies demonstrated the role of Kisspeptin in the final maturation of the oocyte. Cyclooxygenase-2 (Cox-2) has an important enzymatic role in cumulus expansion and follicle rupture. Cox-2 inhibitors, known as an inhibitor of ovulation and follicle rupture, has been shown to selectively suppress KISS1 expression in rat ovaries. All these data give rise to the thought that kisspeptin expression may be associated with Cox-2 expression. In this study, it is aimed to reveal the role of Kisspeptin, Kisspeptin receptor (Kiss1R) and Cox-2, on the oocyte maturation by evaluating the expression levels of Kiss1, Kiss1R and Cox-2 in immature oocytes and cumulus cells, as well as their concentration levels in serum and follicle fluids. In this study, immature and mature oocytes, cumulus cells and follicular fluids obtained from controlled ovarian stimulation cycles and serum from patients were examined. Kiss1, Kiss1R and Cox-2 levels were evaluated in oocytes and cumulus cells by real-time polymerase chain reaction, and in serum and follicle fluid by ELISA. The results were evaluated as the correlation to each other and to the clinical data of the patients. The levels of Kiss1, KissR1 and Cox-2 concentration were shown to not correlate with oocytes maturation level in serum and follicular fluid. While a decrease in the expression of Kiss1 and its receptor was seen in immature oocytes and cumulus cells, an increase in Cox-2 expression was observed. The results of this study suggested that, in controlled ovarian stimulation cycles, Kiss1 and its receptors are effective in oocytes maturation process at the level of microenvironment, and that the negative effects of Cox-2 may control the final maturation step before ovulation.
Description
Keywords
Kisspeptin, Kisspeptin reseptörü, Siklooksijenaz-2, Final oosit matürasyonu, Kisspeptin receptor, Cyclooxygenase-2, Final oocyte maturation
Citation
Kuşpınar, G. (2016). Farklı nüklear matürasyon aşamalarında oosit, kumulus hücreleri ve follikül sıvısında eksprese edilen matürasyon faktörlerinin rolü. Yayınlanmamış yüksek lisans tezi. Uludağ Üniversitesi Sağlık Bilimleri Enstitüsü.