Concentration Assay:Article Title: Estrogen's Effects on Excitatory Synaptic Transmission Entail Integrin and TrkB Transactivation and Depend Upon β 1-integrin function
Article Snippet: Images were processed through restorative deconvolution (99% confidence, Volocity 4.0; Perkin-Elmer) and individual z-stacks were used to construct a three-dimensional (3D) montage of each sample field (136 × 105 × 2 μ m 3 ); elements meeting size constraints of synapses, and detected across multiple intensity thresholds, were quantified using automated systems ( Rex et al , 2009 ; Seese et al , 2013 ). .. Agents used included estrogen receptor- α (ER α ) antagonist MPP (1, 3-Bis (4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy) phenol]-1 H -pyrazole dihydrochloride), ER β antagonist PHTPP (4-[2-phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5- a ]pyrimidin-3-yl]phenoland), β -E2 ((17 β )-estra-1,3,5(10)-triene-3,17-diol), GM6001 and batimastat (Tocris, Bristol, UK), and G-15 (Chemical, Ann Arbor, MI); these compounds were prepared in 100% dimethylsulfoxide and diluted to a final concentration in ACSF. .. TrkB-Fc Chimera Protein (688-TK; R&D Systems, Minneapolis, MN), ANA-12 (Tocris), and neutralizing anti- β 1-integrin (MAB1987Z; Millipore, Billerica, MA) were diluted in ACSF to 2 μ g/ml, 500 nM, and 0.2 mg/ml, respectively.
Article Title: Local synthesis of estradiol in the rostral ventromedial medulla protects against widespread muscle pain in male mice
Article Snippet: .. 864 865 866 Pharmacologic Target Company Cat. # Concentration Solution ICI 182,780 ER antagonist; GPER agonist Tocris Bioscience 1047 0.2–20 μM 0.03% ethanol; 0.9% saline Letrozole Aromatase inhibitor Selleckchem S1235 350 uM–3.5 mM 2% DMSO; 0.9% saline MPP ER-α antagonist Tocris bioscience 1991 10–300 nM 2% DMSO; 0.9% saline PHTPP ER-β antagonist Tocris Bioscience 2662 1 μM 2% DMSO; 0.9% saline eN eu ro Ac ce pte d M an us cri pt 35 869 870 36 Drug Limb Effect for group p value ICI 182,780 (MALE) Ipsilateral Contralateral F4,30 = 1.36 F4,30 = 5.6 p = 0.271 p = 0.002 ICI 182,780 (FEMALE) Ipsilateral Contralateral t9 = 0.15 t9 = 1.22 p = 0.882 p = 0.254 Letrozole Ipsilateral Contralateral F4,19 = 0.09 F4,19 = 8.0 p = 0.985 p = 0.0006 PHTPP Ipsilateral Contralateral t7 = 0.52 t7 = 0.10 p = 0.619 p = 0.922 MPP Ipsilateral Contralateral F2,15 = 2.09 F2,15 = 13.78 P = 0.195 P = 0.0004 ICI182,780 Tukey post-hoc Veh vs 0.2 μM p = 0.051 Veh vs 2 μM p = 0.033 Veh vs 20 μM p = 0.004 Veh vs missed site p = 0.998 Missed site vs 20 μM p = 0.032 Letrozole doses Tukey post-hoc Veh vs 350 μM p = 0.327 Veh vs 1 mM p = 0.022 Veh vs 3.5 mM p = 0.001 Veh vs missed site p = 0.982 Missed site vs 3.5 mM p = 0.003 MPP doses Tukey post-hoc Control vs 300 nM p = 0.0006 Control vs 10 nM p = 0.958 10 nM vs 300 nM p = 0.004 872 Drug Dose Sample size ICI 182,780 (Male) Vehicle Missed site 0.2 uM 2 uM 20 uM 8 5 7 7 8 Letrozole (Male) Vehicle Missed site 350 uM 1 mM 3.5 mM 5 5 4 5 5 eN eu ro Ac ce pte d M an us cri pt 37 876 eN eu ro Ac ce pte d M an us cri pt 38 Gene name F DFn, DFd p value Gabra1 F2, 15 = 1.56 p = 0.243 Gabra2 F2, 15 = 0.65 p = 0.536 Gabrb2 F2, 15 = 2.37 p = 0.128 Gabrb3 F2, 15 = 3.77 p = 0.047 Gabrg1 F2, 15 = 1.06 p = 0.371 Gabrg2 F2, 15 = 1.52 p = 0.249 Oprm1 F2, 15 = 1.51 p = 0.253 Grin1 F2, 15 = 1.77 p = 0.204 Slc32a1 F2, 15 = 0.06 p = 0.938 Oprk1 F2, 15 = 1.10 p = 0.358 877 878 Table 4: Statistical analysis results for quantitative polymerase chain reaction (One-way 879 ANOVA comparing pain-free, sham, and gonadectomized male mice). ..
Saline:Article Title: Local synthesis of estradiol in the rostral ventromedial medulla protects against widespread muscle pain in male mice
Article Snippet: .. 864 865 866 Pharmacologic Target Company Cat. # Concentration Solution ICI 182,780 ER antagonist; GPER agonist Tocris Bioscience 1047 0.2–20 μM 0.03% ethanol; 0.9% saline Letrozole Aromatase inhibitor Selleckchem S1235 350 uM–3.5 mM 2% DMSO; 0.9% saline MPP ER-α antagonist Tocris bioscience 1991 10–300 nM 2% DMSO; 0.9% saline PHTPP ER-β antagonist Tocris Bioscience 2662 1 μM 2% DMSO; 0.9% saline eN eu ro Ac ce pte d M an us cri pt 35 869 870 36 Drug Limb Effect for group p value ICI 182,780 (MALE) Ipsilateral Contralateral F4,30 = 1.36 F4,30 = 5.6 p = 0.271 p = 0.002 ICI 182,780 (FEMALE) Ipsilateral Contralateral t9 = 0.15 t9 = 1.22 p = 0.882 p = 0.254 Letrozole Ipsilateral Contralateral F4,19 = 0.09 F4,19 = 8.0 p = 0.985 p = 0.0006 PHTPP Ipsilateral Contralateral t7 = 0.52 t7 = 0.10 p = 0.619 p = 0.922 MPP Ipsilateral Contralateral F2,15 = 2.09 F2,15 = 13.78 P = 0.195 P = 0.0004 ICI182,780 Tukey post-hoc Veh vs 0.2 μM p = 0.051 Veh vs 2 μM p = 0.033 Veh vs 20 μM p = 0.004 Veh vs missed site p = 0.998 Missed site vs 20 μM p = 0.032 Letrozole doses Tukey post-hoc Veh vs 350 μM p = 0.327 Veh vs 1 mM p = 0.022 Veh vs 3.5 mM p = 0.001 Veh vs missed site p = 0.982 Missed site vs 3.5 mM p = 0.003 MPP doses Tukey post-hoc Control vs 300 nM p = 0.0006 Control vs 10 nM p = 0.958 10 nM vs 300 nM p = 0.004 872 Drug Dose Sample size ICI 182,780 (Male) Vehicle Missed site 0.2 uM 2 uM 20 uM 8 5 7 7 8 Letrozole (Male) Vehicle Missed site 350 uM 1 mM 3.5 mM 5 5 4 5 5 eN eu ro Ac ce pte d M an us cri pt 37 876 eN eu ro Ac ce pte d M an us cri pt 38 Gene name F DFn, DFd p value Gabra1 F2, 15 = 1.56 p = 0.243 Gabra2 F2, 15 = 0.65 p = 0.536 Gabrb2 F2, 15 = 2.37 p = 0.128 Gabrb3 F2, 15 = 3.77 p = 0.047 Gabrg1 F2, 15 = 1.06 p = 0.371 Gabrg2 F2, 15 = 1.52 p = 0.249 Oprm1 F2, 15 = 1.51 p = 0.253 Grin1 F2, 15 = 1.77 p = 0.204 Slc32a1 F2, 15 = 0.06 p = 0.938 Oprk1 F2, 15 = 1.10 p = 0.358 877 878 Table 4: Statistical analysis results for quantitative polymerase chain reaction (One-way 879 ANOVA comparing pain-free, sham, and gonadectomized male mice). ..
Article Title: Local Synthesis of Estradiol in the Rostral Ventromedial Medulla Protects against Widespread Muscle Pain in Male Mice
Article Snippet: MPP , ER-α antagonist , Tocris Bioscience , 1991 , 10–300 nM , 2% DMSO; 0.9% saline. .. PHTPP , ER-β antagonist , Tocris Bioscience , 2662 , 1 μM , 2% DMSO; 0.9% saline. ..
Control:Article Title: Local synthesis of estradiol in the rostral ventromedial medulla protects against widespread muscle pain in male mice
Article Snippet: .. 864 865 866 Pharmacologic Target Company Cat. # Concentration Solution ICI 182,780 ER antagonist; GPER agonist Tocris Bioscience 1047 0.2–20 μM 0.03% ethanol; 0.9% saline Letrozole Aromatase inhibitor Selleckchem S1235 350 uM–3.5 mM 2% DMSO; 0.9% saline MPP ER-α antagonist Tocris bioscience 1991 10–300 nM 2% DMSO; 0.9% saline PHTPP ER-β antagonist Tocris Bioscience 2662 1 μM 2% DMSO; 0.9% saline eN eu ro Ac ce pte d M an us cri pt 35 869 870 36 Drug Limb Effect for group p value ICI 182,780 (MALE) Ipsilateral Contralateral F4,30 = 1.36 F4,30 = 5.6 p = 0.271 p = 0.002 ICI 182,780 (FEMALE) Ipsilateral Contralateral t9 = 0.15 t9 = 1.22 p = 0.882 p = 0.254 Letrozole Ipsilateral Contralateral F4,19 = 0.09 F4,19 = 8.0 p = 0.985 p = 0.0006 PHTPP Ipsilateral Contralateral t7 = 0.52 t7 = 0.10 p = 0.619 p = 0.922 MPP Ipsilateral Contralateral F2,15 = 2.09 F2,15 = 13.78 P = 0.195 P = 0.0004 ICI182,780 Tukey post-hoc Veh vs 0.2 μM p = 0.051 Veh vs 2 μM p = 0.033 Veh vs 20 μM p = 0.004 Veh vs missed site p = 0.998 Missed site vs 20 μM p = 0.032 Letrozole doses Tukey post-hoc Veh vs 350 μM p = 0.327 Veh vs 1 mM p = 0.022 Veh vs 3.5 mM p = 0.001 Veh vs missed site p = 0.982 Missed site vs 3.5 mM p = 0.003 MPP doses Tukey post-hoc Control vs 300 nM p = 0.0006 Control vs 10 nM p = 0.958 10 nM vs 300 nM p = 0.004 872 Drug Dose Sample size ICI 182,780 (Male) Vehicle Missed site 0.2 uM 2 uM 20 uM 8 5 7 7 8 Letrozole (Male) Vehicle Missed site 350 uM 1 mM 3.5 mM 5 5 4 5 5 eN eu ro Ac ce pte d M an us cri pt 37 876 eN eu ro Ac ce pte d M an us cri pt 38 Gene name F DFn, DFd p value Gabra1 F2, 15 = 1.56 p = 0.243 Gabra2 F2, 15 = 0.65 p = 0.536 Gabrb2 F2, 15 = 2.37 p = 0.128 Gabrb3 F2, 15 = 3.77 p = 0.047 Gabrg1 F2, 15 = 1.06 p = 0.371 Gabrg2 F2, 15 = 1.52 p = 0.249 Oprm1 F2, 15 = 1.51 p = 0.253 Grin1 F2, 15 = 1.77 p = 0.204 Slc32a1 F2, 15 = 0.06 p = 0.938 Oprk1 F2, 15 = 1.10 p = 0.358 877 878 Table 4: Statistical analysis results for quantitative polymerase chain reaction (One-way 879 ANOVA comparing pain-free, sham, and gonadectomized male mice). ..
Article Title: Phytoestrogen Agathisflavone Ameliorates Neuroinflammation-Induced by LPS and IL-1β and Protects Neurons in Cocultures of Glia/Neurons
Article Snippet: .. To induce neuroinflammation, at 26 DIV, cocultures were treated for 24 h with LPS (1 μg/mL; Sigma Aldrich, Saint Luis, MO, USA, L2880) or IL-1β (10 ng/mL; R&D Systems, Minneapolis, MN, USA, 501-RL-010); then, the medium was removed and replaced with medium containing just agathisflavone (0.1 or 1 μM) or vehicle, and cultures were analyzed after 24 h. To assess whether the effects of agathisflavone were mediated through ERs, neuron–glial cocultures were treated with the selective ER-α antagonist MPP dihydrochloride at 10 nM (1,3-bis(4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy)phenol]-1H-pyrazole dihydrochloride; Sigma Aldrich, Saint Luis, MO, USA, M7068) or the selective ER-β antagonist PHTPP at 1 μM (4-[2-phenyl-5,7-bis(trifluoromethyl) pyrazolo[1,5-a]pyrimidin-3-yl]phenol; Tocris, Bristol, UK, #2662); control cultures were treated with DMSO vehicle. ..
Real-time Polymerase Chain Reaction:Article Title: Local synthesis of estradiol in the rostral ventromedial medulla protects against widespread muscle pain in male mice
Article Snippet: .. 864 865 866 Pharmacologic Target Company Cat. # Concentration Solution ICI 182,780 ER antagonist; GPER agonist Tocris Bioscience 1047 0.2–20 μM 0.03% ethanol; 0.9% saline Letrozole Aromatase inhibitor Selleckchem S1235 350 uM–3.5 mM 2% DMSO; 0.9% saline MPP ER-α antagonist Tocris bioscience 1991 10–300 nM 2% DMSO; 0.9% saline PHTPP ER-β antagonist Tocris Bioscience 2662 1 μM 2% DMSO; 0.9% saline eN eu ro Ac ce pte d M an us cri pt 35 869 870 36 Drug Limb Effect for group p value ICI 182,780 (MALE) Ipsilateral Contralateral F4,30 = 1.36 F4,30 = 5.6 p = 0.271 p = 0.002 ICI 182,780 (FEMALE) Ipsilateral Contralateral t9 = 0.15 t9 = 1.22 p = 0.882 p = 0.254 Letrozole Ipsilateral Contralateral F4,19 = 0.09 F4,19 = 8.0 p = 0.985 p = 0.0006 PHTPP Ipsilateral Contralateral t7 = 0.52 t7 = 0.10 p = 0.619 p = 0.922 MPP Ipsilateral Contralateral F2,15 = 2.09 F2,15 = 13.78 P = 0.195 P = 0.0004 ICI182,780 Tukey post-hoc Veh vs 0.2 μM p = 0.051 Veh vs 2 μM p = 0.033 Veh vs 20 μM p = 0.004 Veh vs missed site p = 0.998 Missed site vs 20 μM p = 0.032 Letrozole doses Tukey post-hoc Veh vs 350 μM p = 0.327 Veh vs 1 mM p = 0.022 Veh vs 3.5 mM p = 0.001 Veh vs missed site p = 0.982 Missed site vs 3.5 mM p = 0.003 MPP doses Tukey post-hoc Control vs 300 nM p = 0.0006 Control vs 10 nM p = 0.958 10 nM vs 300 nM p = 0.004 872 Drug Dose Sample size ICI 182,780 (Male) Vehicle Missed site 0.2 uM 2 uM 20 uM 8 5 7 7 8 Letrozole (Male) Vehicle Missed site 350 uM 1 mM 3.5 mM 5 5 4 5 5 eN eu ro Ac ce pte d M an us cri pt 37 876 eN eu ro Ac ce pte d M an us cri pt 38 Gene name F DFn, DFd p value Gabra1 F2, 15 = 1.56 p = 0.243 Gabra2 F2, 15 = 0.65 p = 0.536 Gabrb2 F2, 15 = 2.37 p = 0.128 Gabrb3 F2, 15 = 3.77 p = 0.047 Gabrg1 F2, 15 = 1.06 p = 0.371 Gabrg2 F2, 15 = 1.52 p = 0.249 Oprm1 F2, 15 = 1.51 p = 0.253 Grin1 F2, 15 = 1.77 p = 0.204 Slc32a1 F2, 15 = 0.06 p = 0.938 Oprk1 F2, 15 = 1.10 p = 0.358 877 878 Table 4: Statistical analysis results for quantitative polymerase chain reaction (One-way 879 ANOVA comparing pain-free, sham, and gonadectomized male mice). ..
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![Estrogen receptor (ER) activation is required for agathisflavone to inhibit microgliosis and promote remyelination. (A) Root mean square deviation (RMSD) values and rod representation of crystallographic ligand pose (lilac) and the best pose of this ligand generated by DOCK 6.8 (yellow) for each complex. Distances less than 2 Å between the calculated pose and the crystallographic pose indicates that the program was successful in reproducing the experimental data (B) Representation of interactions between agathisflavone (FAB) and retinoic and estrogen receptors; the captions are described in the figure. (C–G) Organotypic cerebellar slices from SOX10-EGFP animals were maintained for 7DIV, then exposed to LPC for 15–17 h, followed by 2 h pretreatment with the selective ER-α antagonist MPP dihydrochloride at 10 nM (1,3-Bis(4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy)phenol]-1H-pyrazole-dihydrochloride), or the selective ER-β antagonist <t>PHTPP</t> at 1 μM (4-[2-Phenyl-5,7-bis(trifluoromethyl) pyrazolo[1,5- a ]pyrimidin-3-yl]phenol), which were kept together with 10 μM FAB for a further 2 DIV. (C) Oligodendrocytes were identified by the Sox10-EGFP reporter (green) and slices were immunolabeled for MBP (red), neurofilament (blue) and Iba-1 (yellow); scale bar: 20 μm. Bar graphs compare LPC and LPC + FAB 10 μM with the effects of the ER antagonists MPP and PHTPP on the NF + axon index (D), the percentage of MBP+/NF + myelinated axons (E), the number of Sox10-EGFP + oligodendrocytes (F) and the number of Iba1+ microglia (G); data are expressed as the mean ± SEM (n = 5); ‡ p < 0.05, ‡‡ p < 0.01, ‡‡‡‡ p < 0.0001 (comparing LPC-DMSO to other treatment groups); †† p < 0.01 and ††† p <0.001 (comparing LPC + FAB10 to LPC + FAB10+MPP); & p < 0.05 (comparing LPC+FAB10+MPP to LPC+FAB10+PHTPP); One-way ANOVA followed by Tukey’s post-hoc test. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_2432/pmc07482432/pmc07482432__gr8.jpg)