Genotoxic Effects of Environmental Pollution in Meriç Delta Fish: Expressions of Antioxidant Defence Signals, Heat Shock Proteins and DNA Damage-Repair Mechanisms

Abstract

In this study, the effects of pollution on economically important fish species for Gala and Sığırcı Lakes [Carassius gibelio (Bloch, 1782), Carassius carassius (Linnaeus, 1758), Scardinius erythrophthalmus (Linnaeus, 1758), Cyprinus carpio Linnaeus, 1758, Sander lucioperca (Linnaeus, 1758), Perca fluviatilis Linnaeus, 1758)] were investigated. The genotoxic effects of environmental pollution were investigated with reference to gene expressions of antioxidant defense signal (GS, Cu-Zn SOD, CAT), heat shock proteins (HSP26, HSP60, HSP70 and HSP83) and DNA repair (SMUG1) by qRT-PCR assay in muscle, liver and gill tissue of fish species. Additionally, DNA damage determined using RAPD assay. Band changes were clearly detected, especially in the gill tissues of all fishes. In addition, a significant increase in the relative expression of CuZn-SOD in muscle tissue of C. gibellio, S. erythrophthalmus and S. lucioperca. The expression of CAT changed significantly at all tissue of fishes. However, the highest increase in expression level of CAT 128.9, 102.1 and 138.4 fg/µg total RNA was determined in muscle, liver and gill tissues of S. lucioperca. While the HSP60 and HSP70 was the least affected by environmental pollutions, greatest DNA repair activity was clearly observed in liver tissue of S. lucioperca. We conclude that environmental pollution in Gala and Sığırcı Lake exert genotoxic effects on DNA stability, and gen expressions of defense signaling. Additionally, S. lucioperca may be a bio-indicator species for biomonitoring to water pollution.

Keywords

• Meriç Delta,fish tissues,RAPD analysis,antioxidant enzymes,gene expression

Meriç Delta Balıklarında Çevre Kirliliğinin Genotoksik Etkileri: Antioksidan Savunma, Isı Şok Protein Sinyali ve DNA Hasar-Onarım Mekanizmaları

Öz

Çalışmamızda, Gala ve Sığırcı Gölleri için ekonomik açıdan önemli olan balık türleri (Carassius gibelio (Bloch, 1782), Carassius carassius (Linnaeus, 1758), Scardinius erythrophthalmus (Linnaeus, 1758), Cyprinus carpio Linnaeus, 1758, Sander lucioperca (Linnaeus, 1758), Perca fluviatilis Linnaeus, 1758) üzerinde kirliliğin etkileri araştırılmıştır. Kirliliğin genotoksik etkileri, kas, solungaç ve karaciğer dokularında antioksidan savunma sinyali (GS, Cu-Zn SOD, CAT), ısı şok proteinleri (HSP26, HSP60, HSP70 ve HSP83) ve QRT-PCR testi ile DNA onarımı (SMUG1) metotları kullanılarak incelenmiştir. Ayrıca, RAPD analizi kullanılarak balık dokularındaki DNA hasarları belirlenmiştir. Bant değişimleri, özellikle tüm balıkların solungaç dokularında açıkça tespit edilmiştir. Ayrıca, C. gibellio, S. erythrophthalmus ve S. lucioperca'nın kas dokusunda CuZn-SOD'un nispi ekspresyonunda önemli bir artış gösterdiği tespit edilmiştir. CAT ekspresyonu tüm balık dokularında önemli ölçüde değişiklik sergilemiştir. Bununla birlikte, CAT 128,9, 102,1 ve 138,4 fg/ug total RNA'nın ekspresyon seviyesindeki en yüksek artış, S. lucioperca'nın kas, karaciğer ve solungaç dokularında belirlenmiştir. HSP60 ve HSP70 çevre kirliliğinden en az etkilenirken, S. lucioperca'nın karaciğer dokusunda en büyük DNA onarım aktivitesi açık şekilde gözlenmiştir. Çalışmamızın sonucunda, Gala ve Sığırcı Göllerindeki çevre kirliliğinin DNA stabilitesi ve savunma sinyallerinin gen ifadeleri üzerinde genotoksik etkileri olduğu ortaya konulmuştur. Ayrıca, S. lucioperca türünün, su kirliliğinin biyolojik olarak izlenmesinde biyoindikatör bir tür olarak kullanılabileceği sonucuna varılmıştır.

Anahtar Kelimeler

• Meriç Deltası,balık dokuları,RAPD analizi,antioksidan enzimler,gen ekspresyonu

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