FIGURE 5.

Acute glucocorticoid treatment attenuates β3 adrenergic signaling but does not affect non-adrenergic ACTH-signaling. (A) Gene expression of melanocortin 2 receptor (Mc2r) in response to varying ambient temperatures obtained from RNA sequencing of murine brown adipose tissue. Data are shown as RPKM (reads per kilobase per million mapped reads) (B) Primary brown adipocytes were either not pretreated or with dexamethason (Dexa) for 1 h prior to microplate-based respirometry. Subsequently, stimulated respiration as fold increase of basal respiration was determined in response to adrenocorticotropic hormone (ACTH), isoproterenol (ISO), Dexa or a combination of ACTH and Dexa. After either no or 1, 2 or 4 h of pretreatment with Dexa (C) ACTH- and ISO-stimulated respiration, relative expression of (D) uncoupling protein 1 (Ucp1), (E) β3-adrenergic receptor (Adrb3) and (F) Mc2r or (G) relative cellular cyclic aenosine monophosphate (cAMP) was assessed in primary brown adipocytes. Data are presented as means ± SD (n = 4). (B,C,G) were analyzed by two-way ANOVA (Tukey’s test). (D–F) were analyzed by one-way ANOVA. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001.