(A) A schematic of the ovarioles that make up a Drosophila ovary (above) and the structure of a single ovariole (below) showing the germline stem cells (left) and a string of increasingly mature follicles. Stages 8, 10, and 14 follicles are labeled. Prophase I arrested oocytes undergo meiotic resumption at stage 10, progressing to a secondary arrest point at metaphase I which is maintained until ovulation. (B) DAPI stained ovaries from females that were fed 1 day (left), fed 1 day then protein restricted for 1 day (middle), or fed 1 day then protein-restricted 13 days (right). Oocytes are colored as in A, revealing the stable storage of two mature stage 14 oocytes per ovariole. Each mature follicle is about 450 μM in length. (C) Eggs laid per day by females containing stored stage 14 follicles, that were provided with males after 3 (green), 6 (blue), 9 (orange) or 12 (red) days. Mating stimulates deposition of the stored oocytes as fertilized embryos. (D) Aging curves (days) for follicles stored in vivo at 29°C (magenta), 25°C (green) or 20°C (blue). For each point, stored oocytes were recovered as in (C) and the hatch rate determined (N > 100). (E) Protein content of mature oocytes that were unstored, stored in vivo for 1 day or for 13 days (‘oocyte age’). Protein restriction of mothers does not affect the protein content of stored mature oocytes. (F) Hatch rate of embryos developing from fresh mature (stage 14) oocytes from 5-day-old females, mature oocytes stored 14 days (during days 2–16) from 16 day-old females, or fresh mature oocytes from 20-day-old females. Oocyte age during storage, but not maternal age, is associated with reduced hatch rate. Error bars in (C), (E), and (F) denote SD.