Differential gene expression in anterior pituitary glands from anestrous and cycling postpartum beef cows

Oligonucleotide microarrays (GeneChip Bovine Genome Arrays, Affymetrix Inc., Santa Clara, CA) were used to evaluate gene expression profiles in anterior pituitary glands collected from 4 anestrous and 4 cycling postpartum primiparous beef cows to provide insight into genes associated with transition from an anestrous to a cycling status. Tissues were collected 40 to 61 d after calving from anestrous cows and from cyclic cows between d 7 and 13 of the estrous cycle (luteal phase) from d 54 to 77 after calving. Hybridization signals were normalized across arrays, and genes with mean differences in expression that were at least 1.5-fold apart and with a minimum difference in mean signal intensity of 10 were compared. Based on these criteria, 47 transcripts were increased (P < 0.025) and 31 transcripts were decreased (P < 0.025) in pituitary gland tissue from cycling compared with anestrous cows. Few transcripts identified in this analysis were associated previously with reproductive function. To provide greater detail on the influence that stage of the estrous cycle (i.e., collection during the luteal phase) might have on the differences detected in gene expression, quantitative real-time PCR was used to compare gene expression in anterior pituitaries of anestrous cows with an additional independent set of anterior pituitary glands collected at 4 different stages of the estrous cycle: 0.5 to 2 d (n = 9), 5 to 6.5 d (n = 5), 11.4 to 13.7 d (n = 5), and 17.9 to 19 d (n = 6) after the onset of estrus. Gastrin-releasing peptide, the gene that exhibited the largest fold increase in expression in the microarray experiment, and IGFBP3 mRNA were expressed at greater (P < 0.004) amounts in samples from the different stages of the estrous cycle than in samples from anestrous cows. In addition, expression of IGFBP3 mRNA was proportional to serum progesterone concentrations throughout the estrous cycle (P < 0.05). Expression of versican mRNA was decreased (P = 0.03) in samples from the different stages of the estrous cycle compared with anestrous cow samples. Results identified numerous genes that may be involved in the transition from anestrous to cycling status, providing novel insight into mechanisms regulating reproductive function.