Adipose tissue (AT) is known to express genes involved in steroid synthesis and metabolism. Due to the ability of dairy cows to mobilize large amounts of body fat, elevated mobilization of AT might increase the release of steroidogenic compounds into the circulation, affecting the physiological steroid hormone balance. In the present study, we tested the mRNA expression of enzymes catalyzing the conversion of cholesterol to 11-deoxycorticosterone i.e., steroidogenic acute regulatory protein (StAR), 3b-hydroxysteroid dehydrogenase (3bHSD), and P450–21-hydroxylase (CYP21) in subcutaneous (sc)AT of dairy cows with different body condition score (BCS). German Holstein cows (n = 38) were preselected and allocated to groups 15 wk before calving, based on either a high (HBCS; BCS >3.75) or normal (NBCS; BCS <3.5) BCS. In addition, back fat thickness (BFT) of both groups was measured (HBCS: BFT >1.4; NBCS: BFT <1.2). Biopsies from scAT (tail head region) were sampled on d −49, 3, 21, and 84 relative to calving. The mRNA abundance of the target genes was assessed by qPCR and normalized by using the 4 most stable reference genes. Data were analyzed using the mixed-model procedure followed by Bonferroni correction (SPSS 21). Differences in BCS and BFT (P < 0.001) due to early grouping of cows were maintained throughout the whole study. From calving until d 84 after calving, HBCS cows lost twice as much BFT compared with NBCS cows (P ≤ 0.001). From d −49 to +84, in both groups the mRNA abundance of CYP21 and StAR increased by 65% (P < 0.01) and 275% (P < 0.001), respectively. The abundance of CYP21 mRNA tended to be 48% higher in NBCS cows on d 21 (P = 0.068) compared with HBCS cows, whereas that of 3bHSD was consistently low over all time points. Increased mRNA abundance of StAR over time, revealed the increased availability of cholesterol for the steroidogenic response. The post-partal increase of CYP21 abundance indicated an enhanced conversion of progesterone into corticosterone in bovine AT. However, the low and unchanged expression of 3bHSD over time gives no indication for a major role of bovine AT in the synthesis of androgens, estrogens and corticosteroids.