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Using an experimental model based on the modulation of follicle growth, this work aimed to evaluate if the polyamine metabolic pathway is regulated by the periovulatory endocrine milieu. Nelore cows were manipulated to ovulate small (n\u2009=\u200915) or large (n\u2009=\u200915) follicles, then the profiles of polyamines and their synthetic enzymes were compared between groups. Transcripts for the enzymes of this pathway, ornithine decarboxylase 1 (ODC1; the rate\u2010limiting enzyme in polyamine biosynthesis) protein quantification, adenosylmethionine decarboxylase 1 (AMD1) protein immunolocalization, and concentrations of the different polyamines (putrescine, spermidine, and spermine) were respectively quantified by quantitative reverse\u2010transcriptase PCR, immunoblotting, immunohistochemistry, and gas chromatography\u2010mass spectrometry in both the endometrium and uterine flushing. No differences in gene and protein expression or concentration of polyamines were observed between groups. There were significant correlations between the relative abundance of <jats:italic>ODC1<\/jats:italic> and spermidine\/spermine N1\u2010acetyltransferase 1 (<jats:italic>SAT1<\/jats:italic>) transcripts as well as between antizyme inhibitor 1 (<jats:italic>AZIN1<\/jats:italic>) and adenosylmethionine decarboxylase 1 (<jats:italic>AMD1<\/jats:italic>) transcripts. In conclusion, our results show that the polyamine metabolic pathway is present and functional, but not regulated by the periovulatory endocrine milieu in the bovine endometrium. <jats:italic>Mol. Reprod. 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