The role of adrenergic activation on murine luteal cell viability and progesterone production Jing Wang, Min Tang, Huaide Jiang, Bing Wu, Wei Cai, Chuan.

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The role of adrenergic activation on murine luteal cell viability and progesterone production Jing Wang, Min Tang, Huaide Jiang, Bing Wu, Wei Cai, Chuan Hu, Riqiang Bao, Qiming Dong, Li Xiao, Gang Li, Chunping Zhang Theriogenology Volume 86, Issue 5, Pages 1182-1188 (September 2016) DOI: 10.1016/j.theriogenology.2016.04.008 Copyright © 2016 Elsevier Inc. Terms and Conditions

Fig. 1 The influence of NE and ISO on murine luteal cell viability. (A) MTT assay for the cultured luteal cells with control, 1-μM NE, 10-μM NE, and 100-μM NE after 2-day culture. (B) MTT assay for the cultured luteal cells with control, 1-μM ISO, 10-μM ISO, and 100-μM ISO after 2-day culture. * P < 0.05. (C) The primary luteal cells with control, 100-μM NE, and 100-μM ISO after 2-day culture. Scale bar, 50 μm. (D) The apoptosis of luteal cells after treatment with control, 100-μM NE, and 100-μM ISO after 2-day culture. Scale bar, 100 μm. ISO, isoprenaline; MTT, (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NE, norepinephrine. Theriogenology 2016 86, 1182-1188DOI: (10.1016/j.theriogenology.2016.04.008) Copyright © 2016 Elsevier Inc. Terms and Conditions

Fig. 2 The effect of propranolol on murine luteal cell viability induced by NE and ISO. (A) MTT assay for the cultured luteal cells with control, 100-μM NE, 10-μM propranolol, and 100-μM NE + 10-μM propranolol after 2-day culture. (B) MTT assay for the cultured luteal cells with control, 100-μM ISO, 10-μM propranolol, and 100-μM ISO + 10-μM propranolol after 2-day culture. * P < 0.05. ISO, isoprenaline; MTT, (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NE, norepinephrine. Theriogenology 2016 86, 1182-1188DOI: (10.1016/j.theriogenology.2016.04.008) Copyright © 2016 Elsevier Inc. Terms and Conditions

Fig. 3 The expression of genes in luteal cells was assessed by Real-time polymerase chain reaction. (A) shows that CyclinD2 messenger RNA (mRNA) was increased and Caspase3 mRNA was decreased after treatment with NE. (B) shows that Caspase3 mRNA was decreased and PGF2α was increased after treatment with ISO. * P < 0.05. ISO, isoprenaline; NE, norepinephrine; PGF2α, prostaglandin F2α. Theriogenology 2016 86, 1182-1188DOI: (10.1016/j.theriogenology.2016.04.008) Copyright © 2016 Elsevier Inc. Terms and Conditions

Fig. 4 The effect of NE and ISO on estradiol and progesterone secretion in luteal cells. The estradiol and progesterone was measured using radioimmunoassay. (A) and (B) showed the estradiol and progesterone content in cultured media, respectively, after treatment with 100-μM NE and 100-μM ISO. (C) showed that the effect of propranolol on progesterone production induced by NE and ISO. * P < 0.05. ISO, isoprenaline; NE, norepinephrine. Theriogenology 2016 86, 1182-1188DOI: (10.1016/j.theriogenology.2016.04.008) Copyright © 2016 Elsevier Inc. Terms and Conditions

Fig. 5 The messenger RNA (mRNA) and protein level of steroidogenic-related enzymes in luteal cells was assessed by real-time polymerase chain reaction and Western blotting. (A) showed the mRNA level of LHR, StAR, and 3β-HSD in luteal cells after treatment with 100-μM NE and 100-μM ISO. (B) showed the protein level of StAR and 3β-HSD in luteal cells after treatment with 100-μM NE and 100-μM ISO. * P < 0.05. 3β-HSD, 3β-hydroxysteroid dehydrogenase; ISO, isoprenaline; NE, norepinephrine. Theriogenology 2016 86, 1182-1188DOI: (10.1016/j.theriogenology.2016.04.008) Copyright © 2016 Elsevier Inc. Terms and Conditions