Autophagy in Frog Visual Cells in Vitro

Isolated frog retinas were incubated in a medium free of serum and amino acids under dim white incandescent light of 20 lux/m2. After 1, 2, 6, and 9 hr of incubation, six retinas at each time point were fixed for electron microscopic investigation. Histochemical staining of acid phosphatase was performed in control and experimental tissues. Autophagic vacuoles in visual cell inner segments were counted and compared with the incidence of vacuoles in control tissues. The ratio of newly formed: old autophagic vacuoles was assessed in incubated retinas, and the number of autophagic vacuoles per rod cell and per cone cell was evaluated. The results indicated that the number of autophagic vacuoles was significantly increased from 1 to 9 hr of incubation, the ratio of newly formed: old autophagic vacuoles was constant over this period, and the amount of autophagy occurring in rods and cones was similar. In a second group of experiments, retinas were incubated under the same conditions but at two different levels of illumination. One series of retinas was incubated in dim red incandescent light of 5 lux/m2, the other series was incubated at bright white fluorescent light of 300 lux/m2. The total numbers of autophagic vacuoles showed a consistent elevation of 20% in bright white light material as compared wot dim red light material. Autophagic vacuoles per cone were significantly higher in retinas incubated in white light than in retinas incubated in red light. Autophagic vacuoles per rod were about equal in both groups. Our observations indicated that visual cells contain an intracellular mechanism of degradation, which is increased under changed metabolic conditions and modified as a function of different levels of illumination.