Glutamine synthetase in cultured whole retinas from the embryonic chick. Role of protein and RNA syntheses in 4 degrees C storage enhancement.

Glutamine synthetase (GS) activity is enhanced in cultured whole retinas when a 72 h incubation at 37 degrees C is preceded by storage at 4 degrees C for 2-24 h. This enhancement occurs even in the absence of glucocorticoids and is maximal in retinas from 11 to 14 d embryos. In comparison, cortisol-induced increases in retinal GS activity at 37 degrees C are optimal in retinas from 8 to 12 d embryos. This study, using cycloheximide (an inhibitor of protein synthesis) and cordycepin (an inhibitor of RNA synthesis), indicates that both protein and RNA synthesis are required for the 4 degrees C storage enhancement of GS activity. The necessary RNA synthesis occurs within the first 48 h following transfer to 37 degrees C and does not require concomitant protein synthesis. Uridine uptake, but not incorporation into trichloroacetic acid-precipitable material, is increased by initial 4 degrees C storage when compared with whole retina controls incubated at 37 degrees C for the total time. In contrast, both uptake and incorporation of amino acids are increased in 4 degrees C-stored retinas for as long as 72 h subsequent to transfer from 4 to 37 degrees C. This suggests that enhancement GS activity may arise from a combination of elevated general protein synthesis and specific messenger-RNA synthesis following 4 degrees C storage.

Effect of dissociative methods on cortisol binding and glutamyltransferase inducibility in chick embryo retina.

The ability of the isolated embryonic chick retina (12 days) to bind a steroid (cortisol) decreases when the tissue is dissociated; the extent of this decrease depends upon the method of dissociation. Trypsin and mechanical dissociation decreased cortisol binding slightly; papain dissociation essentially eliminated it. Cortisol binding decreased with time in culture in both whole retina and monolayer cultures; this decrease may reflect, in part, a similar development decrease in ovo. Inducibility of glutamine synthetase in whole retinas and retinal monolayers prepared with either trypsin or papain also decreased with time in culture. For whole and trypsin-dissociated retinas, the drop in inducibility correlates with the drop in cortisol-binding capacity. This was not the case for monolayer cultures prepared by papain dissociation.