Preantral follicles (PAF) and early antral follicles (EAF) were isolated from bovine ovaries and classified under a stereomicroscope as atretic or healthy. The atretic follicles were all considered as group I (in vivo atresia), whereas healthy follicles were assigned to five groups (group II, in vivo normal control; group III, in vitro normal control; group IV, in vitro induced atresia; groups V and VI, Lipopolysaccharide (LPS)-induced atresia in vitro). Group I and II follicles were immediately snap-frozen (−70°C) until DNA extraction, whereas group III–VI follicles were incubated (39°C, 5% CO2, 95% air) for periods up to 72 hr under various conditions. Group III follicles were maintained in complete medium (M199, bovine calf serum, sodium pyruvate, epidermal growth factor, insulin, transferrin, sodium selenite, penicillin, streptomycin, and amphotericin), whereas group IV follicles were incubated in the same medium, but without serum. Group V and VI follicles were maintained in complete medium, but in the presence of LPS (10 or 50 μ/ml, respectively). Results showed that follicles incubated in the absence of serum and those exposed to both doses of LPS became atretic. DNA isolated from all atretic follicles showed fragmentation typical of that described for apoptosis; this was also confirmed by in situ DNA labeling and histology. Atretic follicles did not produce estradiol (P < 0.001), but progesterone values increased with follicle size (P < 0.001) and time of incubation (P < 0.001). We concluded that in the absence of serum or in the presence of LPS, follicles undergo atresia via apoptosis. © 1996 Wiley-Liss, Inc.