Abstract

On a global scale, the frequencies and magnitudes of hypoxic events in coastal and estuarine waters have increased dramatically over the past 20 years. Fish populations are suitable indicators for the assessment of the quality of aquatic ecosystems, as they are omnipresent and often comprise a variety of different lifestyles and adaption strategies. We have investigated on the molecular level the impact of hypoxia on two fish species typical of European estuaries. We monitored the expression of eleven putatively hypoxia-responsive genes by means of quantitative real-time RT-PCR in brains, gills and hearts of the ruffe (Gymnocephalus cernua) and the flounder (Platichthys flesus). We first investigated the effect of naturally occurring hypoxia in the Elbe estuary. In a second approach, expression changes in the response to hypoxia were monitored under controlled laboratory conditions. The genes that showed the strongest effect were two respiratory proteins, myoglobin and neuroglobin, as well as the apoptosis enzyme caspase 3. As previously observed in other fish, myoglobin, which was considered to be muscle-specific, was found in brain and gills as well. Comparison of field and laboratory studies showed that – with the exception of the heart of flounder – that mRNA levels of the selected genes were about the same, suggesting that laboratory conditions reflect natural conditions. Likewise, trends of gene expression changes under hypoxia were the same, although hypoxia response was more pronounced in the Elbe estuary. In general, the flounder displayed a stronger response to hypoxia than the ruffe, suggesting that the flounder is more susceptible to hypoxia. The most pronounced differences were found among tissues within a species, demonstrating that hypoxia response is largely tissue-specific. In summary, our data suggest that laboratory experiments essentially mimic field data, but additional environmental factors enhance hypoxia response in nature.

Highlights

  • Environmental hypoxia is an increasing problem worldwide with severe consequences for aquatic ecosystems

  • We investigated the molecular response to hypoxia in the ruffe and in the flounder by Quantitative real-time RT-PCR (qRT-PCR) employing a set of genes that represent a broad range of cellular functions

  • Evaluation of the mRNA levels of the three putative reference genes revealed that RPLP0 expression was not affected by the hypoxic conditions in both species

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Summary

Introduction

Environmental hypoxia is an increasing problem worldwide with severe consequences for aquatic ecosystems. In shallow coastal areas and estuaries, environmental hypoxia has been increasing severely over the past decades [3]. These habitats are characterised by various additional anthropomorphic stressors, such as hydro-morphological changes and excessive nutrient inputs as well as natural variations in turbidity and temperature, which might have synergistic impacts [4,5]. Hypoxia mainly occurs during the summer months when the solubility of oxygen in water decreases due to the rising temperature. This effect is further enhanced by the stratification of the water column by formation of thermo- and haloclines [7]. In combination with an insufficient vertical mixing rate through stratification, this leads to bottom-water hypoxia, which is common in estuaries [6,9,10,11]

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