Erformed with Microsoft Excel 2010 and R version 2.11.1.hTAAR9_fwd2: GCATATGAATTCATGGTGAACAATTTCTCCCAAG hTAAR9_rv2: GCTATCAGTAATGTTTTTAATCTGTCTCTACTTCTTCStatisticsFor electrophysiological BMS-790052 dihydrochloride web measurements and reporter gene assays, statistical analysis and curve fitting was done by the Hill equation using Microsoft Excel 2010 or SigmaPlot V8.0 (Systat Software, San Jose, CA). Error bars represent SEM.ChemicalsAll tested aminergic substances and standard chemicals were from Sigma Aldrich or J.T. Baker and dissolved as 100 mM stocks ` in Frog-Ringers solution with the exception of odorants in DMSO. Chemicals used for the initial MedChemExpress CTX-0294885 screening of hTAAR5 were: trimethylamine, tetramethylammonium hydroxide, triethylamine, triethanolamine, tetraethylammonium chloride, b-phenylethylamine, methylamine, dimethylamine, dimethylbutylamine, dimethylethylamine, dimethylisopropylamine, taurine, diethylmethylamine, tyramine, trimethylamine N-oxide, carnosine, octopamine, c-aminobutyric acid, histamine, cyclohexylamine, serotonin, ethanolamine, acetylcholine, choline, aminoacetophenone, N-methylpiperidine, isobutylamine, tropine, pyridine, piperine, tropane, methylmorpholine, dimethylpyrazine, 2-isobutyl-3methoxypyrazine, (2)-nicotine, (+)-nicotine, cotinine, trimethylphosphine, skatole (3-methylindole), putrescine (1,4-diaminobutane), spermidine, cadaverine, muscone, v-pentadecalactone, globalide, galaxolide, aurelione, and Henkel 100 (Henkel, Dusseldorf, Germany) [16]. ?Oligonucleotides1) hTAAR1_fwd: GCGCGGCCGCACCATGATGCCCTTTTGCCACAATATAATTAATAT hTAAR1_rv: GCGGCGGCCGCTGAACTCAATTCCAAAAATAATTTACACC hTAAR2_fwd: GCATATGAATTCATGTATTCATTTATGGCAGGAT hTAAR2_rv: GCATATGCGGCCGCCTACTCACTTTCTTTTTGCATACAC hTAAR5_fwd: GCTATCTATGCTGCATTTGATTTTCAGG hTAAR5_rv: GCTATCATTGAATGTGGGGAGTGCT hTAAR6_fwd: GCATATGAATTCATGAGCAGCAATTCATCCCTGC hTAAR6_rv: GCATATGCGGCCGCTTATATATGTTCAGAAAACAAATTCATG hTAAR8_fwd: GCATATGAATTCATGACCAGCAATTTTTCCCA hTAAR8_rv: GCATATGCGGCCGCTTATTCTAAAAATAAACTAATGGTTGATGA two overlapping fragments hTAAR9_fwd1: CAGAAGATAAACTAACACACAAGA hTAAR9_rv1: GCATATGCGGCCGCAATAAATTAGTTGTTGACGAATCAGTSupporting InformationFigure S1 Evaluation of cell-surface hTAAR5 receptor expression. Expression of the rhodopsin-tagged hTAAR5 receptor in transfected HANA3A cells was detected by immunocytochemical live-cell staining, using the anti-rhodopsin antibody 4D2 and a secondary antibody labeled with the fluorescent dye Alexa Fluor 488 (green). Cell nuclei were stained by DAPI (blue). Scaling bar: 10 mm. (TIF) Figure S2 Concentration response curve of mTAAR5.2)3)4)5)Responses to TMA were normalized to the response to forskolin (10 mM). Calculated EC50 for mTAAR5 is 940 nM. At the same time we repeated measurements for hTAAR5 and were able to reproduce previously calculated EC50 around 100 mM (n = 4). Data are given as mean 6 SEM of 2? independent experiments, each performed in duplicates. Error bars represent SEM. (TIF)AcknowledgmentsWe acknowledge the technical assistance of J. Gerkrath and A. Stoeck.6)Author ContributionsConceived and designed the experiments: GG TH HH. Performed the experiments: IW JK LW SZ AS JA CB MW. Analyzed the data: GG IW JK MW. Wrote the paper: GG IW JK.Human TAAR5 Is Activated by Trimethylamine
Boron neutron capture therapy (BNCT) is based on the nuclear capture and fission reactions of the 10B nucleus with low energy thermal/epithermal neutrons to yield high linear energy transfer a particles and recoiling 7Li nuclei. Since the path lengths of the particles ar.Erformed with Microsoft Excel 2010 and R version 2.11.1.hTAAR9_fwd2: GCATATGAATTCATGGTGAACAATTTCTCCCAAG hTAAR9_rv2: GCTATCAGTAATGTTTTTAATCTGTCTCTACTTCTTCStatisticsFor electrophysiological measurements and reporter gene assays, statistical analysis and curve fitting was done by the Hill equation using Microsoft Excel 2010 or SigmaPlot V8.0 (Systat Software, San Jose, CA). Error bars represent SEM.ChemicalsAll tested aminergic substances and standard chemicals were from Sigma Aldrich or J.T. Baker and dissolved as 100 mM stocks ` in Frog-Ringers solution with the exception of odorants in DMSO. Chemicals used for the initial screening of hTAAR5 were: trimethylamine, tetramethylammonium hydroxide, triethylamine, triethanolamine, tetraethylammonium chloride, b-phenylethylamine, methylamine, dimethylamine, dimethylbutylamine, dimethylethylamine, dimethylisopropylamine, taurine, diethylmethylamine, tyramine, trimethylamine N-oxide, carnosine, octopamine, c-aminobutyric acid, histamine, cyclohexylamine, serotonin, ethanolamine, acetylcholine, choline, aminoacetophenone, N-methylpiperidine, isobutylamine, tropine, pyridine, piperine, tropane, methylmorpholine, dimethylpyrazine, 2-isobutyl-3methoxypyrazine, (2)-nicotine, (+)-nicotine, cotinine, trimethylphosphine, skatole (3-methylindole), putrescine (1,4-diaminobutane), spermidine, cadaverine, muscone, v-pentadecalactone, globalide, galaxolide, aurelione, and Henkel 100 (Henkel, Dusseldorf, Germany) [16]. ?Oligonucleotides1) hTAAR1_fwd: GCGCGGCCGCACCATGATGCCCTTTTGCCACAATATAATTAATAT hTAAR1_rv: GCGGCGGCCGCTGAACTCAATTCCAAAAATAATTTACACC hTAAR2_fwd: GCATATGAATTCATGTATTCATTTATGGCAGGAT hTAAR2_rv: GCATATGCGGCCGCCTACTCACTTTCTTTTTGCATACAC hTAAR5_fwd: GCTATCTATGCTGCATTTGATTTTCAGG hTAAR5_rv: GCTATCATTGAATGTGGGGAGTGCT hTAAR6_fwd: GCATATGAATTCATGAGCAGCAATTCATCCCTGC hTAAR6_rv: GCATATGCGGCCGCTTATATATGTTCAGAAAACAAATTCATG hTAAR8_fwd: GCATATGAATTCATGACCAGCAATTTTTCCCA hTAAR8_rv: GCATATGCGGCCGCTTATTCTAAAAATAAACTAATGGTTGATGA two overlapping fragments hTAAR9_fwd1: CAGAAGATAAACTAACACACAAGA hTAAR9_rv1: GCATATGCGGCCGCAATAAATTAGTTGTTGACGAATCAGTSupporting InformationFigure S1 Evaluation of cell-surface hTAAR5 receptor expression. Expression of the rhodopsin-tagged hTAAR5 receptor in transfected HANA3A cells was detected by immunocytochemical live-cell staining, using the anti-rhodopsin antibody 4D2 and a secondary antibody labeled with the fluorescent dye Alexa Fluor 488 (green). Cell nuclei were stained by DAPI (blue). Scaling bar: 10 mm. (TIF) Figure S2 Concentration response curve of mTAAR5.2)3)4)5)Responses to TMA were normalized to the response to forskolin (10 mM). Calculated EC50 for mTAAR5 is 940 nM. At the same time we repeated measurements for hTAAR5 and were able to reproduce previously calculated EC50 around 100 mM (n = 4). Data are given as mean 6 SEM of 2? independent experiments, each performed in duplicates. Error bars represent SEM. (TIF)AcknowledgmentsWe acknowledge the technical assistance of J. Gerkrath and A. Stoeck.6)Author ContributionsConceived and designed the experiments: GG TH HH. Performed the experiments: IW JK LW SZ AS JA CB MW. Analyzed the data: GG IW JK MW. Wrote the paper: GG IW JK.Human TAAR5 Is Activated by Trimethylamine
Boron neutron capture therapy (BNCT) is based on the nuclear capture and fission reactions of the 10B nucleus with low energy thermal/epithermal neutrons to yield high linear energy transfer a particles and recoiling 7Li nuclei. Since the path lengths of the particles ar.
