Angiogenesis is involved in the local mechanisms that regulate follicular development and ovulation. Recently, the angiopoietin (ANPT)-Tie system has been shown to be required to regulate angiogenesis and blood vessel regression. Expression of the ANPT-Tie system in the cyclic ovary suggests that the relative changes in the expression of ANPT-1 and ANPT-2 influence the stability of ovarian blood vessels. In this study, we investigated 1) the mRNA expression for ANPT-1, ANPT-2, and endothelial cell-specific receptors Tie1 and Tie2 in the theca interna (TI) of the bovine developing, mature, and atretic follicles by using a semiquantitative reverse transcription polymerase chain reaction assay and 2) the effect of ANPT on the secretion of steroid hormones from bovine preovulatory follicles in vitro using a microdialysis system (MDS) implanted in the thecal layer. Bovine follicles were classified as developing, mature, and atretic according to size, follicular fluid content of estradiol (E2) and progesterone (P4), and characteristics of granulosa cells (GCs). Both ANPT and Tie mRNA were expressed in the TI, whereas GCs expressed ANPT mRNA only. The expression of ANPT-2 mRNA was decreased in the mature follicles. This decrease resulted in a decrease in the ANPT-2:ANPT-1 ratio (an index of instability of blood vessels), indicating that the blood vessels became more stable or mature. The early atretic follicles showed a higher ANPT-2:ANPT-1 ratio and higher Tie2 mRNA expression than did other follicles at healthy or later atretic stages. This finding may imply that blood vessels become unstable at the initial stage of follicular atresia. In both mid and late atretic follicles, Tie2 mRNA expression dramatically decreased, indicating a disruption of the ANPT-Tie system. In the MDS experiment, an infusion of ANPT-1 or ANPT-2 increased P4 release, whereas both ANPTs inhibited the release of androstenedione. ANPT-1 also increased E2 release. These results showed that the mRNA expression for ANPT-1, ANPT-2, Tie1, and Tie2 changes during follicular development, maturation, and atresia in bovine follicles and that ANPTs affect steroidogenesis in the preovulatory follicle. The results suggest that the ANPT-Tie system is involved the structural (angiogenesis) and secretory changes that occur during follicular development and atresia.