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BioAssay: AID 2179

Modulation of Metabotropic Glutamate Receptor mGluR4: Rat PAM Fold-Shift

The primary pathophysiological change giving rise to the symptoms of Parkinson's disease (PD) is a loss of the dopaminergic neurons in the substantia nigra pars compacta (SNc) that are involved in modulating the function of basal ganglia (BG) nuclei. Unfortunately, traditional therapies for treatment of PD based on dopamine replacement strategies eventually fail in most patients and are more ..
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 Tested Compounds
 Tested Compounds
All(9)
 
 
Active(7)
 
 
Inactive(2)
 
 
 Tested Substances
 Tested Substances
All(9)
 
 
Active(7)
 
 
Inactive(2)
 
 
AID: 2179
Data Source: Vanderbilt Screening Center for GPCRs, Ion Channels and Transporters (mGluR4 Rat PAM fold shift)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2009-12-07

Data Table ( Complete ):           Active    All
Target
BioActive Compounds: 7
Depositor Specified Assays
AIDNameTypeComment
2437Modulation of the Metabotropic Glutamate Receptor mGluR4summary
2807Modulation of the Metabotropic Glutamate Receptor mGluR4: Calcium Assayconfirmatory
Description:
Assay Provider: Colleen Niswender
Assay Provider Affiliation: Vanderbilt University

The primary pathophysiological change giving rise to the symptoms of Parkinson's disease (PD) is a loss of the dopaminergic neurons in the substantia nigra pars compacta (SNc) that are involved in modulating the function of basal ganglia (BG) nuclei. Unfortunately, traditional therapies for treatment of PD based on dopamine replacement strategies eventually fail in most patients and are associated with numerous side effects. A great deal of effort has been focused on developing a detailed understanding of the circuitry and function of the BG to develop novel, nondopaminergic, approaches for restoring normal BG function in PD patients. Exciting advances suggest that metabotropic glutamate receptors (mGluRs), including the group III mGluRs (mGluR4, -7 and -8), play important roles in regulating transmission through the BG and could serve as targets for novel PD therapeutics (Conn et al., 2005). For instance, mGluR4 activation reduces overactive GABA release at a specific inhibitory BG synapse (Macinnes and Duty, 2008; Marino et al., 2003; Valenti et al., 2003) and reverses motor deficits in a variety of rodent PD models (Konieczny et al., 2007; MacInnes et al., 2004; Marino et al., 2003; Ossowska et al., 2007; Valenti et al., 2003).

To more selectively activate mGluR4 and improve upon the pharmacokinetic liabilities of glutamate analogs, we and others have developed novel positive allosteric modulators (PAMs) which potentiate glutamate function at mGluR4 (Engers et al., 2009; Maj et al., 2003; Marino et al., 2003; Niswender et al., 2008a; Niswender et al., 2008b; Williams et al., 2008); several of these tool compounds exhibit antiparkinsonian and neuroprotective effects in multiple rodent PD models (Battaglia et al., 2006; Marino et al., 2003; Niswender et al., 2008a). Unfortunately, many available compounds have lacked pharmacokinetic properties to make them useful tools for study of mGluR4 function via systemic routes of administration. The probe compound developed here exhibits sufficient potency, efficacy, and pharmacokinetic properties, including brain penetration, to make it a useful compound to progress mGluR4 biology, which will undoubtedly allow the intense study of mGluR4 activation in multiple areas of neuroscience such as psychiatric disorders (Stachowicz et al., 2006; Stachowicz et al., 2004), cancer (Iacovelli et al., 2006), and addiction (Blednov et al., 2004).


References

1. Battaglia G, Busceti CL, Molinaro G, Biagioni F, Traficante A, Nicoletti F and Bruno V (2006) Pharmacological activation of mGlu4 metabotropic glutamate receptors reduces nigrostriatal degeneration in mice treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. J Neurosci 26(27):7222-7229.
2. Blednov YA, Walker D, Osterndorf-Kahanek E and Harris RA (2004) Mice lacking metabotropic glutamate receptor 4 do not show the motor stimulatory effect of ethanol. Alcohol 34(2-3):251-259.
3. Conn PJ, Battaglia G, Marino MJ and Nicoletti F (2005) Metabotropic glutamate receptors in the basal ganglia motor circuit. Nat Rev Neurosci 6(10):787-798.
4. Engers DW, Niswender CM, Weaver CD, Jadhav S, Menon UN, Zamorano R, Conn PJ, Lindsley CW and Hopkins CR (2009) Synthesis and evaluation of a series of heterobiarylamides that are centrally penetrant metabotropic glutamate receptor 4 (mGluR4) positive allosteric modulators (PAMs). J Med Chem 52(14):4115-4118.
5. Iacovelli L, Arcella A, Battaglia G, Pazzaglia S, Aronica E, Spinsanti P, Caruso A, De Smaele E, Saran A, Gulino A, D'Onofrio M, Giangaspero F and Nicoletti F (2006) Pharmacological activation of mGlu4 metabotropic glutamate receptors inhibits the growth of medulloblastomas. J Neurosci 26(32):8388-8397.
6. Konieczny J, Wardas J, Kuter K, Pilc A and Ossowska K (2007) The influence of group III metabotropic glutamate receptor stimulation by (1S,3R,4S)-1-aminocyclo-pentane-1,3,4-tricarboxylic acid on the parkinsonian-like akinesia and striatal proenkephalin and prodynorphin mRNA expression in rats. Neuroscience 145(2):611-620.
7. Macinnes N and Duty S (2008) Group III metabotropic glutamate receptors act as hetero-receptors modulating evoked GABA release in the globus pallidus in vivo. Eur J Pharmacol 580(1-2):95-99.
8. MacInnes N, Messenger MJ and Duty S (2004) Activation of group III metabotropic glutamate receptors in selected regions of the basal ganglia alleviates akinesia in the reserpine-treated rat. Br J Pharmacol 141(1):15-22.
9. Maj M, Bruno V, Dragic Z, Yamamoto R, Battaglia G, Inderbitzin W, Stoehr N, Stein T, Gasparini F, Vranesic I, Kuhn R, Nicoletti F and Flor PJ (2003) (-)-PHCCC, a positive allosteric modulator of mGluR4: characterization, mechanism of action, and neuroprotection. Neuropharmacology 45(7):895-906.
10. Marino MJ, Williams DL, Jr., O'Brien JA, Valenti O, McDonald TP, Clements MK, Wang R, DiLella AG, Hess JF, Kinney GG and Conn PJ (2003) Allosteric modulation of group III metabotropic glutamate receptor 4: a potential approach to Parkinson's disease treatment. Proc Natl Acad Sci U S A 100(23):13668-13673.
11. Niswender CM, Johnson KA, Weaver CD, Jones CK, Xiang Z, Luo Q, Rodriguez AL, Marlo JE, de Paulis T, Thompson AD, Days EL, Nalywajko T, Austin CA, Williams MB, Ayala JE, Williams R, Lindsley CW and Conn PJ (2008a) Discovery, characterization, and antiparkinsonian effect of novel positive allosteric modulators of metabotropic glutamate receptor 4. Mol Pharmacol 74(5):1345-1358.
12. Niswender CM, Lebois EP, Luo Q, Kim K, Muchalski H, Yin H, Conn PJ and Lindsley CW (2008b) Positive allosteric modulators of the metabotropic glutamate receptor subtype 4 (mGluR4): Part I. Discovery of pyrazolo[3,4-d]pyrimidines as novel mGluR4 positive allosteric modulators. Bioorg Med Chem Lett 18(20):5626-5630.
13. Ossowska K, Konieczny J, Wardas J, Pietraszek M, Kuter K, Wolfarth S and Pilc A (2007) An influence of ligands of metabotropic glutamate receptor subtypes on parkinsonian-like symptoms and the striatopallidal pathway in rats. Amino Acids 32(2):179-188.
14. Stachowicz K, Chojnacka-Wojcik E, Klak K and Pilc A (2006) Anxiolytic-like effects of group III mGlu receptor ligands in the hippocampus involve GABAA signaling. Pharmacol Rep 58(6):820-826.
Stachowicz K, Klak K, Klodzinska A, Chojnacka-Wojcik E and Pilc A (2004) Anxiolytic-like effects of PHCCC, an allosteric modulator of mGlu4 receptors, in rats. Eur J Pharmacol 498(1-3):153-156.
15. Valenti O, Marino MJ, Wittmann M, Lis E, DiLella AG, Kinney GG and Conn PJ (2003) Group III metabotropic glutamate receptor-mediated modulation of the striatopallidal synapse. J Neurosci 23(18):7218-7226.
16. Williams R, Niswender CM, Luo Q, Le U, Conn PJ and Lindsley CW (2008) Positive allosteric modulators of the metabotropic glutamate receptor subtype 4 (mGluR4). Part II: Challenges in hit-to-lead. Bioorg Med Chem Lett.
Protocol
Cell culture, plating, and dye loading. HEK/GIRK cells stably expressing the M4 muscarinic receptor were grown in 45 percent Dulbecco's Modified Eagle Media (DMEM), 45 percent Ham's F12, 10 percent fetal bovine serum (FBS), 100 units/ml penicillin/streptomycin, 20 mM HEPES (pH 7.3), 1 mM sodium pyruvate, 2 mM glutamine, and 700 ug/ml G418. The rat mGluR4 cell line was prepared by PCR amplification of the entire coding sequence of each receptor and cloning into pIRES puro 3 (Invitrogen). Cloning sites were BamHI/Not I. HEK/GIRK/M4 cells were transfected with 24 ug of DNA and stable transfectants were selected with puromycin. And a monoclonal cell lines was established. Cells were grown in 45 percent Dulbecco's Modified Eagle Media (DMEM), 45 percent Ham's F12, 10percent fetal bovine serum (FBS), 100 units/ml penicillin/streptomycin, 20 mM HEPES (pH 7.3), 1 mM sodium pyruvate, and 2 mM glutamine (Growth Media). mGluR/GIRK lines were supplemented with 600 ng/ml puromycin dihydrochloride (Sigma-Aldrich) and 700 ug/ml G418 (Mediatech, Inc., Herndon, VA). Cells for experiments were generally maintained for approximately 15-20 passages; this was particularly important for experiments examining the endogenous alpha2C receptor.

Assays were performed within Vanderbilt University's High-Throughput Screening Center. Cells were plated into 384 well, black-walled, clear-bottom poly-D-lysine coated plates (Greiner) at a density of 15,000 cells/20 uL/well in DMEM containing 10 percent dialyzed FBS, 20 mM HEPES, and 100 units/ml penicillin/streptomycin (Assay Media). Plated cells were incubated overnight at 37 degrees C in the presence of 5 percent CO2. The following day, the medium was removed from the cells and 20 uL/well of 1.7 uM concentration of the indicator dye BTC-AM (Invitrogen; prepared as a stock in DMSO and mixed in a 1:1 ratio with pluronic acid F-127) in Assay Buffer (Hanks Balanced Salt Solution (Invitrogen) containing 20 mM HEPES pH 7.3) was added to the plated cells. Cells were incubated for one hour at room temperature and the dye was replaced with 20 uL of Assay Buffer.

Test compound preparation. Glutamate was diluted in Thallium Buffer (125 mM sodium bicarbonate (added fresh the morning of the experiment), 1 mM magnesium sulfate, 1.8 mM calcium sulfate, 5 mM glucose, 12 mM thallium sulfate, 10 mM HEPES, pH 7.3) at 5x the final concentration to be assayed. For fold shift experiments, compounds were diluted to a 60 uM (2x final) concentration in Assay Buffer. Cell plates and compound plates were loaded onto a Hamamatsu FDSS 6000 kinetic imaging plate reader. Appropriate baseline readings were taken (10 images at 1 Hz, excitation, 470+/-20 nm emission, 540+/-30 nm) and test compounds were added. Compounds were added in a 20 uL volume and incubated for approximately 2.5 minutes prior to the addition of 10 ul of Thallium Buffer +/- agonist. After the addition of agonist, data were collected for an additional 2 min.

Thallium sulfate requires special handling and disposal precautions and investigators are cautioned to contact their Environmental Health and Safety Department to ensure proper procedures are followed.

Data analysis. Data were analyzed using usoft Excel. Raw data were opened in Excel and each data point in a given trace was divided by the first data point from that trace (static ratio). For experiments in which antagonists/potentiators were added, data were again normalized by dividing each point by the fluorescence value immediately before the agonist addition to correct for any subtle differences in the baseline traces after the compound incubation period. The slope of the fluorescence increase beginning five seconds after thallium/agonist addition and ending fifteen seconds after thallium/agonist addition was calculated. Curves were fitted using a four point logistical equation using XLfit (IDBS, Bridgewater, NJ). Subsequent confirmations of concentration-response parameters were performed using independent serial dilutions of source compounds and data from multiple days experiments were integrated and fit using a four point logistical equation in GraphPad Prism (GraphPad Software, Inc., La Jolla, CA).

For compounds that changed the EC50 of glutamate 3 fold or less, 'Outcome' was assigned as 'Inactive' and 'Score' was assigned as '1'. For compounds that changed the EC50 of glutamate greater than 3 fold, 'Outcome' was assigned as 'Active'. 'Score' was assigned as '50' for those with a fold-shift less than or equal to 10 or as '100' for those with a fold-shift greater than 10.
Result Definitions
Show more
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Rep1ForExperiment1_0.0000155_uM (1.55e-05μM**)Value for replicate 1 for experiment 1 at 0.0000155 uMFloat
2Rep2ForExperiment1_0.0000155_uM (1.55e-05μM**)Value for replicate 2 for experiment 1 at 0.0000155 uMFloat
3Rep1ForExperiment1_0.000156_uM (0.000156μM**)Value for replicate 1 for experiment 1 at 0.000156 uMFloat
4Rep2ForExperiment1_0.000156_uM (0.000156μM**)Value for replicate 2 for experiment 1 at 0.000156 uMFloat
5Rep1ForExperiment1_0.00156_uM (0.00156μM**)Value for replicate 1 for experiment 1 at 0.00156 uMFloat
6Rep2ForExperiment1_0.00156_uM (0.00156μM**)Value for replicate 2 for experiment 1 at 0.00156 uMFloat
7Rep1ForExperiment1_0.00900_uM (0.009μM**)Value for replicate 1 for experiment 1 at 0.00900 uMFloat
8Rep2ForExperiment1_0.00900_uM (0.009μM**)Value for replicate 2 for experiment 1 at 0.00900 uMFloat
9Rep1ForExperiment1_0.0624_uM (0.0624μM**)Value for replicate 1 for experiment 1 at 0.0624 uMFloat
10Rep2ForExperiment1_0.0624_uM (0.0624μM**)Value for replicate 2 for experiment 1 at 0.0624 uMFloat
11Rep1ForExperiment1_0.394_uM (0.394μM**)Value for replicate 1 for experiment 1 at 0.394 uMFloat
12Rep2ForExperiment1_0.394_uM (0.394μM**)Value for replicate 2 for experiment 1 at 0.394 uMFloat
13Rep1ForExperiment1_2.49_uM (2.49μM**)Value for replicate 1 for experiment 1 at 2.49 uMFloat
14Rep2ForExperiment1_2.49_uM (2.49μM**)Value for replicate 2 for experiment 1 at 2.49 uMFloat
15Rep1ForExperiment1_15.7_uM (15.7μM**)Value for replicate 1 for experiment 1 at 15.7 uMFloat
16Rep2ForExperiment1_15.7_uM (15.7μM**)Value for replicate 2 for experiment 1 at 15.7 uMFloat
17Rep1ForExperiment1_100_uM (100μM**)Value for replicate 1 for experiment 1 at 100 uMFloat
18Rep2ForExperiment1_100_uM (100μM**)Value for replicate 2 for experiment 1 at 100 uMFloat
19Rep1ForExperiment1_1000_uM (1000μM**)Value for replicate 1 for experiment 1 at 1000 uMFloat
20Rep2ForExperiment1_1000_uM (1000μM**)Value for replicate 2 for experiment 1 at 1000 uMFloat
21AverageForExperiment1_0.0000155uMAverage for experiment 1 at 0.0000155 uMFloat
22AverageForExperiment1_0.000156uMAverage for experiment 1 at 0.000156 uMFloat
23AverageForExperiment1_0.00156uMAverage for experiment 1 at 0.00156 uMFloat
24AverageForExperiment1_0.00900uMAverage for experiment 1 at 0.00900 uMFloat
25AverageForExperiment1_0.0624uMAverage for experiment 1 at 0.0624 uMFloat
26AverageForExperiment1_0.394uMAverage for experiment 1 at 0.394 uMFloat
27AverageForExperiment1_2.49uMAverage for experiment 1 at 2.49 uMFloat
28AverageForExperiment1_15.7uMAverage for experiment 1 at 15.7 uMFloat
29AverageForExperiment1_100uMAverage for experiment 1 at 100 uMFloat
30AverageForExperiment1_1000uMAverage for experiment 1 at 1000 uMFloat
31StddevForExperiment1_0.0000155uMStandard deviation for experiment 1 at 0.0000155 uMFloat
32StddevForExperiment1_0.000156uMStandard deviation for experiment 1 at 0.000156 uMFloat
33StddevForExperiment1_0.00156uMStandard deviation for experiment 1 at 0.00156 uMFloat
34StddevForExperiment1_0.00900uMStandard deviation for experiment 1 at 0.00900 uMFloat
35StddevForExperiment1_0.0624uMStandard deviation for experiment 1 at 0.0624 uMFloat
36StddevForExperiment1_0.394uMStandard deviation for experiment 1 at 0.394 uMFloat
37StddevForExperiment1_2.49uMStandard deviation for experiment 1 at 2.49 uMFloat
38StddevForExperiment1_15.7uMStandard deviation for experiment 1 at 15.7 uMFloat
39StddevForExperiment1_100uMStandard deviation for experiment 1 at 100 uMFloat
40StddevForExperiment1_1000uMStandard deviation for experiment 1 at 1000 uMFloat
41GluPotencyExperiment1Glutamate potency for experiment 1Float
42GluFoldShiftExperiment1Glutamate fold shift for experiment 1Float
43Rep1ForExperiment2_0.0000155_uM (1.55e-05μM**)Value for replicate 1 for experiment 2 at 0.0000155 uMFloat
44Rep2ForExperiment2_0.0000155_uM (1.55e-05μM**)Value for replicate 2 for experiment 2 at 0.0000155 uMFloat
45Rep1ForExperiment2_0.000156_uM (0.000156μM**)Value for replicate 1 for experiment 2 at 0.000156 uMFloat
46Rep2ForExperiment2_0.000156_uM (0.000156μM**)Value for replicate 2 for experiment 2 at 0.000156 uMFloat
47Rep1ForExperiment2_0.00156_uM (0.00156μM**)Value for replicate 1 for experiment 2 at 0.00156 uMFloat
48Rep2ForExperiment2_0.00156_uM (0.00156μM**)Value for replicate 2 for experiment 2 at 0.00156 uMFloat
49Rep1ForExperiment2_0.00900_uM (0.009μM**)Value for replicate 1 for experiment 2 at 0.00900 uMFloat
50Rep2ForExperiment2_0.00900_uM (0.009μM**)Value for replicate 2 for experiment 2 at 0.00900 uMFloat
51Rep1ForExperiment2_0.0624_uM (0.0624μM**)Value for replicate 1 for experiment 2 at 0.0624 uMFloat
52Rep2ForExperiment2_0.0624_uM (0.0624μM**)Value for replicate 2 for experiment 2 at 0.0624 uMFloat
53Rep1ForExperiment2_0.394_uM (0.394μM**)Value for replicate 1 for experiment 2 at 0.394 uMFloat
54Rep2ForExperiment2_0.394_uM (0.394μM**)Value for replicate 2 for experiment 2 at 0.394 uMFloat
55Rep1ForExperiment2_2.49_uM (2.49μM**)Value for replicate 1 for experiment 2 at 2.49 uMFloat
56Rep2ForExperiment2_2.49_uM (2.49μM**)Value for replicate 2 for experiment 2 at 2.49 uMFloat
57Rep1ForExperiment2_15.7_uM (15.7μM**)Value for replicate 1 for experiment 2 at 15.7 uMFloat
58Rep2ForExperiment2_15.7_uM (15.7μM**)Value for replicate 2 for experiment 2 at 15.7 uMFloat
59Rep1ForExperiment2_100_uM (100μM**)Value for replicate 1 for experiment 2 at 100 uMFloat
60Rep2ForExperiment2_100_uM (100μM**)Value for replicate 2 for experiment 2 at 100 uMFloat
61Rep1ForExperiment2_1000_uM (1000μM**)Value for replicate 1 for experiment 2 at 1000 uMFloat
62Rep2ForExperiment2_1000_uM (1000μM**)Value for replicate 2 for experiment 2 at 1000 uMFloat
63AverageForExperiment2_0.0000155uMAverage for experiment 2 at 0.0000155 uMFloat
64AverageForExperiment2_0.000156uMAverage for experiment 2 at 0.000156 uMFloat
65AverageForExperiment2_0.00156uMAverage for experiment 2 at 0.00156 uMFloat
66AverageForExperiment2_0.00900uMAverage for experiment 2 at 0.00900 uMFloat
67AverageForExperiment2_0.0624uMAverage for experiment 2 at 0.0624 uMFloat
68AverageForExperiment2_0.394uMAverage for experiment 2 at 0.394 uMFloat
69AverageForExperiment2_2.49uMAverage for experiment 2 at 2.49 uMFloat
70AverageForExperiment2_15.7uMAverage for experiment 2 at 15.7 uMFloat
71AverageForExperiment2_100uMAverage for experiment 2 at 100 uMFloat
72AverageForExperiment2_1000uMAverage for experiment 2 at 1000 uMFloat
73StddevForExperiment2_0.0000155uMStandard deviation for experiment 2 at 0.0000155 uMFloat
74StddevForExperiment2_0.000156uMStandard deviation for experiment 2 at 0.000156 uMFloat
75StddevForExperiment2_0.00156uMStandard deviation for experiment 2 at 0.00156 uMFloat
76StddevForExperiment2_0.00900uMStandard deviation for experiment 2 at 0.00900 uMFloat
77StddevForExperiment2_0.0624uMStandard deviation for experiment 2 at 0.0624 uMFloat
78StddevForExperiment2_0.394uMStandard deviation for experiment 2 at 0.394 uMFloat
79StddevForExperiment2_2.49uMStandard deviation for experiment 2 at 2.49 uMFloat
80StddevForExperiment2_15.7uMStandard deviation for experiment 2 at 15.7 uMFloat
81StddevForExperiment2_100uMStandard deviation for experiment 2 at 100 uMFloat
82StddevForExperiment2_1000uMStandard deviation for experiment 2 at 1000 uMFloat
83GluPotencyExperiment2Glutamate potency for experiment 2Float
84GluFoldShiftExperiment2Glutamate fold shift for experiment 2Float
85Rep2ForExperiment3_1000_uM (1000μM**)Value for replicate 2 for experiment 3 at 1000 uMFloat
86Rep1ForExperiment3_1000_uM (1000μM**)Value for replicate 1 for experiment 3 at 1000 uMFloat
87Rep2ForExperiment3_100_uM (100μM**)Value for replicate 2 for experiment 3 at 100 uMFloat
88Rep1ForExperiment3_100_uM (100μM**)Value for replicate 1 for experiment 3 at 100 uMFloat
89Rep2ForExperiment3_31.6_uM (31.6μM**)Value for replicate 2 for experiment 3 at 31.6 uMFloat
90Rep1ForExperiment3_31.6_uM (31.6μM**)Value for replicate 1 for experiment 3 at 31.6 uMFloat
91Rep2ForExperiment3_10.0_uM (10μM**)Value for replicate 2 for experiment 3 at 10.0 uMFloat
92Rep1ForExperiment3_10.0_uM (10μM**)Value for replicate 1 for experiment 3 at 10.0 uMFloat
93Rep2ForExperiment3_3.16_uM (3.16μM**)Value for replicate 2 for experiment 3 at 3.16 uMFloat
94Rep1ForExperiment3_3.16_uM (3.16μM**)Value for replicate 1 for experiment 3 at 3.16 uMFloat
95Rep2ForExperiment3_1.00_uM (1μM**)Value for replicate 2 for experiment 3 at 1.00 uMFloat
96Rep1ForExperiment3_1.00_uM (1μM**)Value for replicate 1 for experiment 3 at 1.00 uMFloat
97Rep2ForExperiment3_0.316_uM (0.316μM**)Value for replicate 2 for experiment 3 at 0.316 uMFloat
98Rep1ForExperiment3_0.316_uM (0.316μM**)Value for replicate 1 for experiment 3 at 0.316 uMFloat
99Rep2ForExperiment3_0.100_uM (0.1μM**)Value for replicate 2 for experiment 3 at 0.100 uMFloat
100Rep1ForExperiment3_0.100_uM (0.1μM**)Value for replicate 1 for experiment 3 at 0.100 uMFloat
101Rep2ForExperiment3_0.0100_uM (0.01μM**)Value for replicate 2 for experiment 3 at 0.0100 uMFloat
102Rep1ForExperiment3_0.0100_uM (0.01μM**)Value for replicate 1 for experiment 3 at 0.0100 uMFloat
103Rep2ForExperiment3_0.00100_uM (0.001μM**)Value for replicate 2 for experiment 3 at 0.00100 uMFloat
104Rep1ForExperiment3_0.00100_uM (0.001μM**)Value for replicate 1 for experiment 3 at 0.00100 uMFloat
105AverageForExperiment3_1000uMAverage for experiment 3 at 1000 uMFloat
106AverageForExperiment3_100uMAverage for experiment 3 at 100 uMFloat
107AverageForExperiment3_31.6uMAverage for experiment 3 at 31.6 uMFloat
108AverageForExperiment3_10.0uMAverage for experiment 3 at 10.0 uMFloat
109AverageForExperiment3_3.16uMAverage for experiment 3 at 3.16 uMFloat
110AverageForExperiment3_1.00uMAverage for experiment 3 at 1.00 uMFloat
111AverageForExperiment3_0.316uMAverage for experiment 3 at 0.316 uMFloat
112AverageForExperiment3_0.100uMAverage for experiment 3 at 0.100 uMFloat
113AverageForExperiment3_0.0100uMAverage for experiment 3 at 0.0100 uMFloat
114AverageForExperiment3_0.00100uMAverage for experiment 3 at 0.00100 uMFloat
115StddevForExperiment3_1000uMStandard deviation for experiment 3 at 1000 uMFloat
116StddevForExperiment3_100uMStandard deviation for experiment 3 at 100 uMFloat
117StddevForExperiment3_31.6uMStandard deviation for experiment 3 at 31.6 uMFloat
118StddevForExperiment3_10.0uMStandard deviation for experiment 3 at 10.0 uMFloat
119StddevForExperiment3_3.16uMStandard deviation for experiment 3 at 3.16 uMFloat
120StddevForExperiment3_1.00uMStandard deviation for experiment 3 at 1.00 uMFloat
121StddevForExperiment3_0.316uMStandard deviation for experiment 3 at 0.316 uMFloat
122StddevForExperiment3_0.100uMStandard deviation for experiment 3 at 0.100 uMFloat
123StddevForExperiment3_0.0100uMStandard deviation for experiment 3 at 0.0100 uMFloat
124StddevForExperiment3_0.00100uMStandard deviation for experiment 3 at 0.00100 uMFloat
125GluPotencyExperiment3Glutamate potency for experiment 3Float
126GluFoldShiftExperiment3Glutamate fold shift for experiment 3Float
127AvgGluPotency*Average glutamate potencyFloatμM
128StddevGluPotencyStandard deviation for glutamate potencyFloat
129SEMGluPotencySEM for glutamate potencyFloat
130AvgGluFoldShiftAverage glutamate fold shiftFloat
131StddevGluFoldShiftStandard deviation for glutamate fold shiftFloat
132SEMGluFoldShiftSEM for glutamate fold shiftFloat

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: NS053536

Data Table (Concise)
Classification
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