In vitro embryo kültürü ve embryo transferi ile elde edilen farelerde testis dokusunda toll benzeri reseptörlerin ekspresyonu

Abstract

Toll benzeri reseptörlerin (TLR'ler) fare testis dokusunda immun yanıtta rol oynamaktan ziyade gelişimsel rolleri de olabileceğini gösteren kanıtlar mevcuttur. Her ne kadar fonksiyonel TLR proteinlerinin tümü fare testis dokusunda eksprese edilse de, TLR genlerinin transkripsiyonel ekspresyonunu fizyolojik koşullarda kapsamlı bir şekilde araştıran bir çalışmaya rastlanılmamıştır. Ayrıca, fare testis dokusunda TLR transkriptlerinin ekspresyon düzeylerinin in vitro embriyo kültürü ve embriyo transferi işlemlerinden etkilenip etkilenmediği bilinmemektedir. Bu nedenle, mevcut çalışmanın ilk amacı, fare testis dokusunda tüm TLR'lerin transkripsiyonel ekspresyonunu fizyolojik koşullarda incelemektir. Sunulan çalışmanın ikinci amacı ise, atmosferik oksijen koşulları altında kültüre edilen blastosistlerin yalancı gebe farelere transferinden elde edilen yetişkin testis dokularında TLR'lerin transkripsiyonel ekspresyon düzeylerinin, doğal olarak yumurtlayan dişilerden elde edilen yetişkin farelere ait testis dokuları ile kıyaslandıklarında, değişip değişmediğini araştırmaktır. Çalışmada sekiz haftalık Agouti farelerine (C57BL/6 x BALB/c) ait testis dokuları kullanılmıştır. Çalışmada bir adet Kontrol grubu ve bir adet Deneme grubu yer almıştır. Tüm TLR genlerinin ekspreyon düzeylerini belirlemek amacıyla Kantitatif-Gerçek Zamanlı -Polimeraz Zincir Reaksiyonu (qRT-PCR) yönteminden yaralanılmıştır. Elde edilen veriler, tüm TLR transkriptlerinin kontrol ve deneme grubuna ait yetişkin farelerin testis dokularında mevcut olduğunu göstermiştir. Bununla birlikte, TLR transkriptlerinin ekspresyon düzeylerinin gruplar arasında benzer olduğu ortaya konulmuştur. Elde edilen sonuçlar, fare testis dokusunda TLR ekspreyonunun in vitro embryo kültürü ve embryo transferinden etkilenmediğini göstemektedir. Ancak, tüm TLR transkriptlerinin hem fizyolojik ve fizyolojik olmayan koşullar altında testis dokusunda ekspresyonu, TLR'lerin fare spermatogenezinde fizyolojik rollere sahip olabileceğini düşündürmektedir. TLR'ler ile spermatogenezin gelişimsel süreçleri arasındaki ilişkiyi ortaya koymak amacıyla daha fazla araştırmaya ihtiyaç vardır.

Keywords

• Embriyo transferi
• fare
• in vitro embriyo kültürü
• testis
• toll benzeri reseptörler

Supporting Institution

Sunulan çalışma herhangi bir finansal destek almadan gerçekleştirilmiştir.

Ethical Statement

Bu çalışmada kullanılan hayvan materyalleri, Aydın Adnan Menderes Üniversitesi Hayvan Deneyleri Yerel Etik Kurulu tarafından 28.11.2019, 64583101/2019/121 Numaralı Etik Kurul Kararı ile onaylanan projeden elde edilmiştir.

Expression of Toll-Like Receptors in the Mouse Testis Derived From In Vitro Embryo Culture and Embryo Transfer

Abstract

Evidence suggests that Toll-like receptors (TLRs) may play developmental rather than immune roles in the mouse testis. Although all TLR proteins are present in the mouse testis, there are no studies that investigate the transcriptional expression of all TLR genes in physiological conditions. It also remains unexplored whether expression levels of TLR transcripts in mouse testis are influenced by in vitro embryo culture and embryo transfer. Therefore, the first objective of the present study was to examine transcriptional expression of all TLRs in mouse testis in physiological conditions. The second objective was to investigate whether transcriptional expressions of TLRs alter in testes of mice derived from transfer of blastocyst cultured under the atmospheric oxygen conditions, compared to those obtained from naturally ovulated females. Testis tissues of eight-week-old Agouti mice (C57BL/6 x BALB/c) were used in the current study. The study included one Control group (CG) and one Experimental group (EG). Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to assess the expressions of TLR genes. Data indicated that all TLR transcripts were present in the testis of CG and EG mice. Nonetheless, none of the TLR transcripts showed differential expressions between groups. TLR expression in the mouse testis appears to remain unaffected by in vitro embryo culture and embryo transfer. Expression of all TLRs in mouse testis under both physiological and detrimental conditions suggests that TLRs may have physiological roles in the process of spermatogenesis. Further studies are warranted to investigate the connection between TLRs and developmental processes of spermatogenesis.

Keywords

• mouse
• testis
• Embryo transfer
• in vitro embryo culture
• mouse
• testis
• toll-likereceptors

Supporting Institution

The present study was conducted without any financial support.

Ethical Statement

This study was approved by the Aydın Adnan Menderes University Animal Experiments Local Ethics Committee (28.11.2019, 64583101/2019/121 Number Ethics Committee Decision).

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