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We investigated the effects of insulin-like growth factor 1 (IGF-1) on the morphology and follicular activation of ovine preantral follicles cultured in situ and whether the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway is involved in IGF-1 action in the sheep ovary. Ovine ovarian fragments were fixed for histological and terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) analyses (fresh control) or cultured in supplemented alpha-minimum essential medium (α-MEM+; control) or α-MEM+ with IGF-1 (1, 10, 50, 100 or 200 ng mL−1) for 7 days. Follicles were classified as normal or atretic, primordial or growing and the oocyte and follicle diameters were measured. DNA fragmentation was evaluated by TUNEL assay. Proliferating cell nuclear antigen (PCNA) immunohistochemistry was performed on the fresh control, α-MEM+ and 100 ng mL−1 IGF-1 samples. Inhibition of PI3K activity was performed through pretreatment with the PI3K inhibitor LY294002 and phosphorylated AKT (pAKT) expression was analysed after culture in the absence or presence of LY294002. IGF-1 at 100 ng mL−1 increased (P < 0.05) follicular activation compared with α-MEM+ and decreased TUNEL-positive cells (P < 0.05) compared with other treatments. PCNA-positive cells also increased (P < 0.05) in 100 ng mL−1 IGF-1. LY294002 significantly inhibited follicular activation stimulated by α-MEM+ and 100 ng mL−1 IGF-1 and reduced pAKT expression in follicles. Overall, IGF-1 at 100 ng mL−1 promoted primordial follicle activation, cell proliferation and reduced DNA fragmentation after in situ culture through the PI3K/AKT pathway. Primordial follicle growth or activation is under the control of several paracrine factors such as insulin-like growth factor 1 (IGF-1). This study investigated the effects of IGF-I on ovine ovarian tissue culture and it demonstrated that IGF-1 promoted activation through the PI3K/AKT signalling pathway. Controlling the levels of specific signalling members in the follicle culture can lead to retrieval of oocytes with an enhanced developmental potential. [ABSTRACT FROM AUTHOR]