Effects of assay and acclimation temperatures on incorporation of amino acids into protein of isolated hepatocytes from the european eel, L.

Hepatocytes of adult eels acclimated to 5° C, 10° C and 20° C, respectively were isolated by perfusion of the liver with collagenase. The liver-somatic index and the protein content of liver cells showed significantly higher values in fish kept at the lower temperatures. However, in the adenine nucleotide content and energy charge no significant differences were observed between the 5° C and the 20° C acclimation groups. The incorporation of radioactivity from a ¹⁴C-labelled amino acid mixture into perchloric acid precipitates was used as an estimate of over-all protein synthesis. When eel hepatocytes were incubated in Hanks' solution containing tracer amounts of amino acids, labelling of perchloric acid precipitates showed linear time courses over at least 60 min at 10° C and 20° C assay temperatures. The total cellular radioactivity, however, exhibited non-linear time courses. In the measurement range from 5° C to 25° C Arrhenius plots of protein labelling exhibited a discontinuity in both groups of fish. Hepatocytes from 10° C-acclimated eel showed almost twice the incorporation rates of amino acids as those from the 20° C-acclimated fish. It is concluded that high temperature dependencies in the low temperature range require an increase in the capacity of the apparatus for protein synthesis during cold acclimation.