Ractopamine Hydrochloride and Estradiol + Trenbolone Acetate Implants Alter Myogenic mRNA, <i>β</i>-Adrenergic Receptors, and Blood Metabolites

Two commonly used growth promotants in the United States beef industry are β-agonists and anabolic steroid hormones. Each has been shown to increase lean muscle deposition in cattle provided treatments of each growth technology, but much is still unknown of how steroidal implants and β-agonists work in combination. It was our goal to determine the effect of implant strategy and β-agonist administration in beef feedlot heifers (n = 264). A 3 × 2 factorial randomized complete block design was used with 2 levels of OPT and 3 different durations of terminal implant (TI) windows for a total of 6 treatment groups with 9 replications. Terminal implants (20 mg estradiol/200 mg trenbolone acetate implant, Component TE-200) were provided to heifers 140 d from slaughter (TI140), 100 d from slaughter (TI100), or 60 d from slaughter (TI60). Animals receiving the later two TI being first implanted on day 0 (8 mg estradiol/80 mg trenbolone acetate implant, Component TE-IH). The second treatment of the cattle received was the orally active beta adrenergic agonist, ractopamine-hydrochloride (RH) in the form of Optaflexx®(OPT;0 (NO) or 200 (YES) mg/hd·d-1) over the final 28 days of the trial. Thirty animals were subjected to longissimus muscle (LM) biopsies on d 0, 40, 80, 112, and at slaughter on d 140 to view mRNA levels of myogenic related genes and protein quantities of the β1-adrenergic receptor (β1 AR) and β2-adrenergic receptor (β2 AR). On the same days, blood samples were taken from 108 animals to assess changes in plasma blood urea nitrogen (BUN), non-esterified fatty acids (NEFA) and progesterone due to treatments. Relative mRNA levels of myosin heavy chain IIX (MHC IIX), AMPKα, and IGF-I were increased (P < 0.05) in animals receiving a TI100 over the other two implant dates after OPT was fed to animals. After OPT administration myosin heavy chain IIA (MHC IIA) mRNA levels tended to decrease (P = 0.09) due to OPT. An interaction between TI d and OPT administration caused an increase (P < 0.05) in MHC IIA mRNA level in the TI60/Yes treatment group over all other treatments except the TI100/No treatment group. Protein intensity of the β2 AR was decreased (P < 0.05) by the latest TI d (TI60) during OPT feeding, while β1 AR protein intensity tended to be lower (P < 0.10) in animals fed OPT. Plasma BUN levels were reduced (P < 0.05) after terminal implants and OPT feeding;while progesterone was decreased (P < 0.05) by OPT alone. Neither growth promotant affected NEFA levels in plasma. Collectively, these data indicate that ractopamine hydrochloride and estradiol + trenbolone acetate implants alter myogenic mRNA, β-adrenergic receptors, and blood metabolites in finishing beef heifers.

Zinc Alters Ractopamine HCl Response in Cultured Skeletal Muscle Cells

The objective of this study was to determine if zinc, when added in combination with ractopamine hydrochloride (RH), would stabilize the interaction of RH with the β-adrenergic receptor, as indicated by altered cAMP concentrations, mRNA quantity, or protein abundance. Cultured bovine skeletal muscle cells were established and treated after 120 h for 6, 24, and 96 h with differentiation media of specific treatments. Treatments were applied in a factorial arrangement with two levels of zinc (0 μM or 1 μM) and two levels of RH (0 μM or 10 μM) in differentiation media. cAMP levels were measured at 6, 24, and 96 h, while mRNA and protein were measured at 24 and 96 h. At 6 h, no differences (P > 0.05) were detected in cAMP levels between the treatments. However, at 24 h the 10 μM RH, 1 μM zinc treatment had the greatest concentrations of cAMP (P < 0.05). At 96 h the 10 μM RH, 0 μM zinc treatment had a lower concentration of cAMP (P = 0.05) compared to the control. No differences were detected in mRNA (β1-adrenergic receptor, β2-adenergic receptor, AMPKα, myosin heavy chain I, myosin heavy chain IIA, and myosin heavy chain IIX) concentrations between treatments. Protein quantity of the β1-adrenergic receptor and β2-adrenergic receptor did not differ between treatments. These results indicate that zinc, in combination with RH, may help sustain the RH response during prolonged exposure as indicated by increased cAMP concentrations.

Interactive Effects of Zinc and Zilpaterol Hydrochloride on Bovine <i>β</i>-Adrenergic Receptors

The objective of this study was to determine if the addition of zinc (Zn) in combination with zilpaterol HCL (ZH) affected the interaction of ZH with the beta2-adrenergic receptor (β-AR) by altering cAMP production, gene expression, and protein abundance in cultured skeletal muscle cells. Cultures of muscle bovine satellite cells were established and treated at 120 h with: 1) 0 μM Zn/zilpaterol hydrochloride (ZH;CON);2) 0 μM Zn/10 μM ZH (ZH);3) 1 μM Zn from Zn chloride/0 μM ZH (Zn);4) 1 μM Zn from Zn chloride/10 μM ZH (ZN/ZH) in differentiation media for an additional 0, 6, 24, 48 and 96 h. Protein and mRNA were isolated and quantified at 24 and 96 h, and cAMP was measured at 0, 6, 24, 48 and 96 h. At 0, 24, 48 and 96 h, no differences (P > 0.05) were detected in cAMP production. At 6 h, Zn cells had the greatest concentration of cAMP (P < 0.05) compared to ZH treatments. No differences (P > 0.05) were detected in mRNA abundance at 24 h. At 96 h, 0 μM Zn/10 μM ZH cells had an increased abundance of myosin heavy chain (MHC)-I mRNA (P < 0.05) compared to CON. Furthermore, ZH had a greater abundance of MHC-IIX mRNA (P < 0.05) and a tendency for a greater abundance of IGF-1 mRNA (P < 0.15) compared to CON and ZN/ZH. No differences (P > 0.05) were detected in the protein abundance of β1AR and the β2AR. These results indicated Zn and ZH in combination did not have an additive effect on β2-AR function as indicated by cAMP concentrations.