Abstract
Microinjection was tested as a potentially powerful tool to introduce natural and anthropogenic pollutants directly into fish eggs to determine their toxicological impact on fish. With this technique, parental transfer of lipophilic contaminants may be mimicked. Here, we investigated the applicability of pollutant injection into the yolk of early zebrafish (Danio rerio) eggs with special regard on survival after vehicle injection. Tested vehicles were autoclaved tap water, dimethyl sulfoxide (DMSO), methanol, and triolein. Highest mortality occurred after the injection of DMSO and methanol. The lethality rates were up to 40% higher than under control conditions. Best survival rates were obtained after triolein and water injections. However, the triolein droplet was not assimilated by the embryo within 96 h post fertilization suggesting an incomplete uptake of triolein-solubilized chemicals. Technical aspects concerning microinjection in zebrafish eggs are discussed with special emphasis on quantitative injection. Microinjection into the yolk cell of zebrafish eggs is feasible, but the application of exact volumes appears problematic. However, microinjection is a powerful tool for studies without the demand for high volume accuracy. Adopting microinjection for pollutant research requires further investigation.
Highlights
Microinjection was tested as a potentially powerful tool to introduce natural and anthropogenic pollutants directly into fish eggs to determine their toxicological impact on fish
The survival after vehicle injection ranged from 100% to 60% depending on the injected substance as well as the injection volume within 24 h post fertilization
Individuals injected with 4.2 nL dimethyl sulfoxide (DMSO) or methanol showed significantly lower survival rates than zebrafish eggs after injection with similar volumes of autoclaved tap water or triolein (Figure 1A, B, Table 2)
Summary
Microinjection was tested as a potentially powerful tool to introduce natural and anthropogenic pollutants directly into fish eggs to determine their toxicological impact on fish. With this technique, parental transfer of lipophilic contaminants may be mimicked. Life stages of aquatic species including fish face different pollutant exposure routes. Microinjection allows, e.g., the production of transgenic cell lines or animals. It offers the direct administration of supporting or harmful substances into cells to investigate their mode of action or toxic potential [10,11].
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