Gökkuşağı Alabalıklarında (Oncorhynchus mykiss) Kloramin-T Uygulaması Sonucu Solungaçlarda Meydana Gelen Oksidatif Stresin Beyin Dokusundaki c-Fos ve BDNF Gen Ekspresyonu Üzerine Etkisinin Araştırılması

Kloramin-T, balıklarda solungaç hastalıklarını önlemek ve/veya bu hastalıklardan korunmak için kullanılan bir kimyasaldır. Birçok balık türünde solungaçlarda oluşan bakteriyel hastalıklar kloramin-T uygulanarak kontrol altına alınmaktadır. Ancak yanlış kloramin-T uygulaması sucul canlılarda oksidatif strese neden olmaktadır. Bu çalışmada, gökkuşağı alabalıklarına (Oncorhynchus mykiss) uygulanan kloramin-T’nin solungaçlarda oluşturacağı oksidatif stres düzeyinin araştırılması ve bu stresin beyindeki nöronal aktiviteyi etkileyip etkilemediğinin araştırılması amaçlanmaktadır. Bu amaç için gökkuşağı alabalıklarına sub-lethal dozda (2.8 mg/L) 0-1-24-48-72 ve 96 saat kloramin-T uygulaması yapıldı. Daha sonra antioksidant enzimleri kodlayan genlerin ( süperoksit dismütaz 1 (SOD1), süperoksit dismütaz 2 (SOD2), katalaz (CAT), glutathion peroksidaz 1 (GPX1), glutathion peroksidaz 4 (GPX4)) mRNA transkript seviyeleri qRT-PCR ile ölçüldü. Son olarak beyin dokularında nöronal aktiviteyi ölçmek amacıyla c-Fos ve BDNF genlerine ait ekspresyon seviyeleri yine qRT-PCR ile belirlendi. Kloramin-T uygulanan balıkların solungaçlarında SOD1, SOD2, CAT ve GPX1 genlerinin ekspresyon seviyelerinin kontrol grubuna göre önemli derecede arttığı gözlemlendi. Ancak GPX4 geninde önemli bir artış gözlemlenmedi. Aynı balıkların beyin dokuları incelendiğinde c-fos ve BDNF ekspresyonlarının benzer şekilde kontrol grubuna göre artış gösterdiği belirlendi. Çalışmadan elde edilen bu sonuçlar, aşırı ve/veya yanlış kloramin-T uygulamasının gökkuşağı alabalıklarının solungaç dokularında oksidatif strese neden olduğunu ve oksidatif strese maruz kalan balıkların beyin dokularında nöronal aktivitenin negatif yönde etkilendiğini göstermektedir.

Anahtar Kelimeler:

Kloramin-T, oksidatif stres, gökkuşağı alabalığı, c-Fos, BDNF.

Investigation of the Effect of Oxidative Stress in Gill Tissue on the Brain Tissue c-Fos and BDNF Gene Expression after Chloramine-T Application of Rainbow Trout (Oncorhynchus mykiss)

Chloramine-T is a chemical used to prevent and/or protect gill diseases in fish. In many fish species, bacterial diseases that occur in the gills are controlled by chloramine-T application. However, the wrong application of chloramine-T causes oxidative stress in aquatic organisms. In this study, it is aimed to investigate the oxidative stress level of chloramine-T applied to rainbow trout (Oncorhynchus mykiss) in gills and to investigate whether this stress affects neuronal activity in brain. For this purpose, rainbow trout were used sub-lethal dose (2.8 mg / L) 0-1-24-48-72 and 96 hours of chloramine-T. The levels of mRNA transcripts of genes encoding antioxidant enzymes (superoxide dismutase 1 (SOD1), superoxide dismutase 2 (SOD2), catalase (CAT), glutathione peroxidase 1 (GPX1), glutathione peroxidase 4 (GPX4)) were measured by qRT-PCR. Finally, expression levels of c-Fos and BDNF genes were determined by qRT-PCR to measure neuronal activity in brain tissues. It was observed that the expression levels of SOD1, SOD2, CAT and GPX1 genes increased significantly in the gills of fish treated with chloramine-T. However, there was no significant increase in GPX4 gene. When the brain tissues of the same fish were examined, it was determined that c-fos and BDNF expressions increased in comparison to the control group. The results of the study show that excessive and/or incorrect chloramine-T application causes oxidative stress in the gill tissues of rainbow trout and negative effects of neuronal activity on brain tissues of fish exposed to oxidative stress.

Keywords:

Chloramine-T, oxidative stress, rainbow trout, c-Fos, BDNF,

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