Evaluation of ezrin and fascin 1 in the PFOS treated Sertoli cell culture: An in vitro study

Yıl 2020, Cilt: 4 Sayı: 1, 16 - 20, 02.03.2020

Nazlı Ece Gungor-ordueri

https://doi.org/10.30565/medalanya.573983

Öz

Aim: Depending on the findings resulting from the knock-downing of ezrin and of fascin 1 in vivo, we aim to show the defects or disruption of the blood-testis barrier (BTB) structure and F-actin bundling after Perfluorooctanesulfonate (PFOS) treatment in primary Sertoli cell culture. 

Methods: Study Design: Primary Sertoli cell isolation was occurred with control and PFOS-treated (20M) groups. Sertoli cells were prepared for both experiments as 0.5 x 106 cell/ml. Method: Dual-labeled immunofluorescence analysis to assess co-localization of fascin 1 with ezrin both in Sertoli cells was performed, and Co-IP, by using lysates of seminiferous tubules, was performed using actin and ezrin proteins to identify specific protein-protein interaction with fascin 1. 

Results: Firstly, we showed that ezrin and fascin 1, which were components of the ectoplasmic specialization were co-localized in the Sertoli cells and also they were interacted each other. Secondly, we indicated that they were dislocated in the PFOS-treated Sertoli cells in vitro. Because of PFOS (20M), the actin-based cytoskeleton was no longer capable of supporting the distribution and/or localization of actin-regulatory proteins at the cell-cell interface necessary to maintain localization of actin-regulatory at the BTB.

Conclusion: In summary, these findings suggest that ezrin and fascin 1 can work together to preserve BTB integrity by regulating F-actin organization in the PFOS-mediated Sertoli cell disruption. 

Anahtar Kelimeler

Blood-testis barrier, ectoplasmic specialization, F-actin, fascin-1, PFOS

Destekleyen Kurum

The Rockefeller University, The Population Council

Teşekkür

Dr. C. Yan Cheng

PFOS ile tedavi edilen Sertoli hücre kültüründe ezrin ve fascin 1'in araştırılması: İn vitro bir çalışma

Öz

Amaç: Bu çalışmada, Ezrin ve Fascin 1’in in vivo olarak baskılanması sonucundaki bulgularla birlikte, Kan testis bariyeri (KTB) yapısının bozulması ve primer Sertoli hücre kültüründe, Perflorooktansülfonat (PFOS) muamelesinden sonra F-aktin demetlenmesinin gösterilmesi amaçlanmaktadır.

Yöntem: Çalışma Tasarımı: Primer Sertoli hücre izolasyonu, kontrol ve PFOS ile muamele edilen (20μM) gruplarla yapıldı. Sertoli hücre konsantrasyonu her iki deney için 0.5 x 106 hücre/ml olacak şekilde hazırlandı. Metod: Hem Fascin 1'in Sertoli hücrelerinde Ezrin ile birlikte lokalizasyonunu değerlendirmek üzere çift etiketli immünofloresan analizi yapıldı, hem de seminifer tübül lizatlarından aktin ve Ezrin proteinlerinin Fascin 1 proteini ile spesifik protein-protein etkileşimini tanımlamak için Co-IP deneyi uygulandı.

Bulgular: İlk olarak, ektoplazma özelleşmesi bileşenleri olan Ezrin ve Fascin 1 in Sertoli hücrelerinde ko-lokalize olduğu ve birbirleri ile etkileşime girdiği gösterilmiştir. İkinci olarak ise PFOS ile muamele edilen Sertoli hücrelerinde Ezrin ve Fascin 1 dislokasyonu gösterilmiştir. 20μM PFOS uygulandığında aktin tabanlı hücre iskeleti KTB’deki hüce-hücre bağlantı bölgelerindeki aktin düzenleyici proteinlerin yayılmasını ve/veya lokalizasyonunu destekleyememiştir.

Tartışma: Sonuç olarak bu bulgular, PFOS aracılı Sertoli hücre bozulmasında, Ezrin ve Fascin 1'in KTB bütünlüğünü korumak için F-aktin organizasyonunu düzenleyerek birlikte çalışabileceğini desteklemektedir.

Anahtar Kelimeler

Kan-testis bariyeri, ektoplazma özelleşmesi, F-aktin, Ezrin, Fascin-1, PFOS

Kaynakça

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