Intravenous Microinjection of Nephrotoxins into Zebrafish Embryo: A Technique to Deliver Nephrotoxic Agents into the Bloodstream of Zebrafish Embryo

Intravenous microinjection facilitates the delivery of molecules of interest into the bloodstream of a recipient animal. To deliver nephrotoxins into a zebrafish embryo, first, prepare an appropriate concentration of desired nephrotoxin solution.

Using a fine gel loading tip, load this nephrotoxin solution into a microinjection needle with a sharpened angular tip. Suspend the needle vertically to allow the nephrotoxin solution to fill the needle tip by gravity. Now, secure the loaded microinjection needle into the holder of a micromanipulator.

Subsequently, transfer an anesthetized zebrafish embryo into the prefabricated microinjection mold. Align the embryo in the mold with its head oriented within the well and the tail out of the well, improving the accessibility to the tail for microinjection. Next, precisely position the needle tip near the embryo.

Inject the nephrotoxin solution into the tail vein of the embryo. The injected nephrotoxic agents spread through the circulation and reach distant organs like the kidney. Gently retract the needle. Transfer the embryo into a fresh dish containing embryo media and allow it to recover. Incubate the embryo for a specific duration.

Within hours of injection, the nephrotoxins accumulate in the kidney and cause damage, leading to a decline in kidney function.