Glyphosate (GLY) is an herbicidal active ingredient and both in vitro and in vivo studies suggest that GLY alters ovarian function. To determine if a chronic GLY exposure model affected steroidogenesis or folliculogenesis in vivo, postnatal day 42 C57BL6 female mice were orally delivered vehicle control (saline) or GLY (2 mg/Kg) from a pipette tip five days per week for either five or ten weeks. Mice were euthanized at the pro-estrus stage of the estrous cycle. GLY exposure did not impact body weight gain, organ weights, or healthy follicle numbers. In addition, GLY exposure did not affect abundance of ovarian mRNA encoding kit ligand (Kitlg), KIT protooncogene receptor tyrosine kinase (c-Kit), insulin receptor (Insr), insulin receptor substrate (Irs1 or Irs2) and protein thymoma viral proto-oncogene 1 (AKT) or phosphorylated AKT. Ovarian mRNA or protein abundance of Star, 3β-hydroxysteroid dehydrogenase (Hsd3b1), Cyp11a1 or Cyp19a were also not altered by GLY. Circulating 17β-estradiol and progesterone concentration were unaffected by GLY exposure. In conclusion, chronic GLY exposure for five or ten weeks did not affect the ovarian endpoints examined herein.