G-protein coupled receptors (GPR) have been widely studied within human medicine as drug targets for metabolic disorders. They play central roles in many cell signaling processes, and also have application within dairy nutrition as targets for specific dietary components; for example, omega-3 fatty acids. To combat metabolic disorders prevalent in dairy cows during the transition period, which often co-occur with negative energy balance and lipid mobilization, it may be helpful to identify locations, activities, and roles of free fatty acid receptors (FFAR) and other members of the GPR family. To provide insight on tissue-specific differences in gene expression, and therefore, potential downstream pathways of fatty acid-sensing GPR in bovine, quantitative RT-PCR (qPCR) of subcutaneous adipose, liver, and polymorphonuclear leukocyte (PMN) samples during the transition period (−15, +10, and +30 d) were used for expression profiling of medium- (MCFA) and long-chain fatty acid (LCFA) receptors GPR120 and GPR40, MCFA receptor GPR84, and niacin receptor GPR109A. Results were analyzed in SAS to examine differences in expression over time (P < 0.05). In adipose tissue, GPR120 expression was highest at −15 d, decreased at +10 d, and further decreased at +30 d. GPR40 expression was highest at +10 d relative to other time points, and GPR109A expression was lower prepartum than both postpartum time points. GPR84 was undetectable. In liver tissue, GPR84 and GPR109A were nearly undetectable and did not differ (P > 0.05) across time. GPR40 had lower expression at +30 d than −15 or +10 d. GPR120 was undetectable. In PMN, GPR120 expression was increased between +10 and +30 d but neither were significantly different from −15 d. GPR84 expression was higher at +10 d relative to other time points. Neither GPR40 nor GPR109A were altered in PMN. The data suggest that there is likely not a direct role for the selected GPR in the liver during the transition period, but they do play variable roles in adipose and PMN. Dietary supplementation or exclusion of LCFA and/or niacin may provide a method of regulating GPR function during the prepartal or early postpartal periods.