Abstract
We have developed and validated a novel technique that allows for a relatively quick and efficient isolation of embryonic tissue from the yolk in the rainbow trout, Oncorhynchus mykiss. After removal of the chorion, the yolk is first dissolved in an imidazole-KCl buffer and separated from the embryo proper and yolk sac (tissues) by centrifugation. SDS gel was used to identify proteins of 165, 48, 35 and 10 kDa unique to the embryonic tissue and a 6 kDa protein unique to the yolk. Using protein analysis and enzyme activity measurements (lactate dehydrogenase and glutamate dehydrogenase) of each fraction, we have determined that cross contamination is less than 10%. This new method will facilitate biochemical studies of the embryo and yolk that were previously very time consuming.
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Shahsavarani, A., Thomas, Z., Ballantyne, J. et al. A novel technique for the separation of yolk from the developing embryonic tissue in a teleost fish, Oncorhynchus mykiss . Fish Physiology and Biochemistry 24, 321–326 (2001). https://doi.org/10.1023/A:1015074405382
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DOI: https://doi.org/10.1023/A:1015074405382
- early development
- embryo
- glutamate dehydrogenase
- lactate dehydrogenase
- rainbow trout
- yolk protein
- yolk sac