This study aimed to determine the effects of rumen-protected methionine and choline supplementation during the transition period on mRNA expression of follicular cells of the 1st postpartum dominant follicle in Holstein cows. Multiparous cows were assigned in a randomized complete block design into 4 treatments from 21 d before calving to 30 DIM. Treatments were: CON (n = 10, fed the close-up and fresh cow diets with a Lys:Met = 3.5:1), MET (n = 9, fed the basal diet + methionine, Smartamine M to a Lys:Met = 2.9:1), CHO (n = 9, fed the basal diets + choline 60 g/d, Reashure), and MIX (n = 12, fed the basal diets plus Smartamine M to a Lys:Met = 2.9:1 and 60 g/d Reashure). Follicular development was monitored via ultrasound every 2 d starting at 7 DIM until the first dominant follicle reached a diameter of 16 mm. Follicular fluid from each cow was aspirated and cells were retrieved immediately by centrifugation and stored at −80°C until RNA extraction. Statistical analysis was performed using the MIXED procedure of SAS. Gene expression of LHCGR, STAR, 3β-HSD, P450scc, P450c17, CYP19A1, IRS1, IGF, MAT1A, SAHH, TLR4, TNF, IL1-β, IL8 and IL6 was measured by real-time PCR. Treatments did not affect mRNA expression of LHCGR, STAR, P450scc, CYP19A, SAHH, MAT1A and IL6 (P > 0.05) however, 3β-HSD expression was higher (P < 0.05) for MET (1.46 ± 0.3) and MIX (1.25 ± 0.3) than CON (0.17 ± 0.04) and CHO (0.26 ± 0.1). For TNF, TLR4 and IL1-B mRNA expression was higher (P < 0.05) for CON (11.70 ± 4.6, 21.29 ± 10.4, 6.28 ± 1.4) than CHO (2.77 ± 0.9, 2.16 ± 0.9, 2.29 ± 0.7) and MIX (2.23 ± 0.7, 1.46 ± 0.6, 2.92 ± 0.8). There was higher (P < 0.05) IL1-β expression and a tendency (P = 0.07) for higher TNF expression in CON (6.27 ± 1.4, 11.70 ± 4.6) than MET (3.28 ± 0.6, 3.06 ± 0.8). There was no difference (P = 0.43) between CON and MET for TLR4. Expression of IL8 mRNA was lower (P < 0.05) for CHO (0.98 ± 0.3) than CON (4.90 ± 0.7), MET (6.10 ± 1.7) and MIX (5.05 ± 1.8). In conclusion, supplementing Smartamine M and Reashure during the transition period changed mRNA expression in follicular cells of the 1st postpartum dominant follicle in Holstein cows.