Glyphosate (GLY) usage for weed control is extensive. To investigate ovarian impacts of chronic GLY exposure, female C57BL6 mice were orally administered saline as vehicle control (CT) or GLY at 0.25 (G0.25), 0.5 (G0.5), 1.0 (G1.0), 1.5 (G1.5), or 2 (G2.0) mg/kg for five days per wk for 20 wks. Feed intake increased (P < 0.05) in G1.5 and G2.0 mice and body weight increased (P < 0.05) in G1.0 mice. There was no impact of GLY on estrous cyclicity, nor did GLY affect circulating levels of 17β-estradiol or progesterone. Exposure to GLY did not impact heart, liver, spleen, kidney or uterus weight. Both ovarian weight and follicle number were increased (P < 0.05) by G2.0 but not affected at lower GLY concentrations. There were no detectable effects of GLY on ovarian protein abundance of pAKT, AKT, pAKT:AKT, H2AX, STAR, CYP11A1, HSD3B, CYP19A, ERA or ERB. Increased (P < 0.05) abundance of ATM protein was observed at G0.25 but not higher GLY doses. A dose-dependent effect (P < 0.10) of GLY exposure on ovarian protein abundance as quantified by LC-MS/MS was observed (G0.25 – 4 increased, 19 decreased; G0.5 – 5 increased, 25 decreased; G1.0 - 65 increased, 7 decreased; G1.5 - 145 increased, 2 decreased; G2.0 – 159 increased, 4 decreased). Pathway analysis was performed using DAVID and identified glutathione metabolism, metabolic and proteasome pathways as GLY exposure targets. These data indicate that chronic low-level exposure to GLY alters the ovarian proteome and may ultimately impact ovarian function.