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BioAssay: AID 623888

Modulation of mGlu3 (mGlu3_GIRK_Schild)

The hippocampus is a limbic cortical structure that plays an important role in a number of normal physiological processes and is a primary site of pathology in certain neurological disorders, such as Alzheimer's disease and temporal lobe epilepsy (see (Brown and Zador 1990) for review). Because of its important role in both normal and pathological processes, a great deal of effort has been more ..
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 Tested Substances
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Active(1)
 
 
AID: 623888
Data Source: Vanderbilt Specialized Chemistry Center (mGlu3_Thallium_Flux_Schild)
BioAssay Type: Panel
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2012-03-27
Hold-until Date: 2013-03-27
Modify Date: 2013-03-27

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compound: 1
Related Experiments
AIDNameTypeProbeComment
602451Optimization of novel mGluR3-selective allosteric modulatorsSummary depositor-specified cross reference: Optimization of novel mGluR3-selective allosteric modulators
623885Modulation of the Metabotropic Glutamate Receptor mGluR3 (mGlu3 GIRK Potency)Confirmatory2 same project related to Summary assay
623929Modulation of the Metabotropic Glutamate Receptor mGluR3 (mGlu2 GIRK Potency)Confirmatory same project related to Summary assay
623931ML289 Competition in Radioligand Binding assays (Ricerca)Other same project related to Summary assay
624028Modulation of the Metabotropic Glutamate Receptor mGluR3 (mGlu3 Galpha15 Calcium Potency)Confirmatory1 same project related to Summary assay
651839Modulation of the Metabotropic Glutamate Receptor mGluR3 (Negative Allosteric Modulators, mGlu3 GIRK Potency)Confirmatory5 same project related to Summary assay
651840Modulation of the Metabotropic Glutamate Receptor mGluR3 (Negative Allosteric Modulators mGlu3 Galpha15 Calcium Potency)Confirmatory15 same project related to Summary assay
Description:
The hippocampus is a limbic cortical structure that plays an important role in a number of normal physiological processes and is a primary site of pathology in certain neurological disorders, such as Alzheimer's disease and temporal lobe epilepsy (see (Brown and Zador 1990) for review). Because of its important role in both normal and pathological processes, a great deal of effort has been focused on developing a detailed understanding of the cellular mechanisms involved in regulation of transmission through the hippocampal circuit. Evidence suggests that the metabotropic glutamate receptors (mGlus) play a variety of roles in modulating transmission and cell excitability at each of the major excitatory synapses in the hippocampus (Anwyl 1999; Coutinho and Knopfel 2002). In particular, the Gi/o-coupled group II mGlus (mGlu2 and mGlu3) can have significant physiological effects in the hippocampus. Activation of group II mGlus reduces transmission at perforant path-dentate gyrus synapses (Macek et al. 1996), the mossy fiber synapse (Nicholls et al. 2006), and at synapses onto certain interneuron populations (Doherty et al. 2004). However, although presynaptic group II mGlus do not directly reduce synaptic transmission through actions on glutamatergic terminals at the SC-CA1 synapse (Gereau and Conn 1995b; Winder et al. 1996; Fitzjohn et al. 1999), we and others previously reported extensive studies demonstrating that group II mGlus in this region are involved in a novel form of glial-neuronal communication. In particular, activation of group II mGlus induces a marked potentiation of cAMP responses elicited by activation of beta-adrenergic receptors (betaARs) in astrocytes, leading to release of adenosine from the astrocytes and inducing a profound depression of transmission at the SC-CA1 synapse via activation of presynaptic A1 adenosine receptors (Winder et al. 1996; Moldrich et al. 2002). This unique response provides a mechanism by which activation of group II mGlus only reduces transmission at the SC-CA1 synapses under conditions where betaARs and group II mGlus are coincidentally activated. This could provide a protective mechanism to reduce risk of excitotoxicity during periods in which there is excessive excitatory drive when noradrenergic inputs are highly active, such as during periods of intense or prolonged stress.

Unfortunately, despite concerted efforts by multiple groups, selective reagents that differentiate between mGlu2 and mGlu3 have not been available to allow determination of the specific mGlu subtype involved in modulating betaAR-mediated cAMP responses and effects on synaptic transmission. However, within our mGlu5 program, we have discovered compounds with mGlu3 NAM activity, such as VU0092273 (Rodriguez et al. 2010), that have excellent aqueous solubility and achieve high CNS exposure. However, this compound also acts as an mGlu5 PAM. While the mGlu5 PAM activity prevents use of VU0092273 as a selective mGluR3 NAM probe, discovery of this compound still represents a major breakthrough in that it is the first compound that selectively blocks mGlu3 relative to mGlu2 and establishes the feasibility of optimizing novel molecules that act at mGlu3 without effects on the closely related mGlu2 subtype. Thus, it will now be critical to further optimize compounds based on VU0092273 that are selective for mGlu3 relative to all other mGlu subtypes. Establishing novel, highly selective mGlu3 probes represents a critical need that could directly impact our understanding of mGlu3 function and could have a major impact on the direction of current drug discovery efforts focused on discovery and development of agents that activate or inhibit group II mGlus.

This unique response provides a mechanism by which activation of group II mGlus only reduces transmission at the SC-CA1 synapses under conditions where betaARs and group II mGlus are coincidentally activated. This could provide a protective mechanism to reduce risk of excitotoxicity during periods in which there is excessive excitatory drive when noradrenergic inputs are highly active, such as during periods of intense or prolonged stress. However, it is also possible that this reduction in transmission at a key hippocampal synapse could impair hippocampal function and could contribute to the known disrupting effects of stress on hippocampal synaptic plasticity and cognitive function (Howland and Wang 2008). If so, selective antagonists of the specific group II mGlu subtype that mediates this response also have potential utility as cognition-enhancing agents. Consistent with this, while non-selective group II mGlu agonists can reduce stress responses (Swanson et al. 2005) these agents also impair hippocampal-dependent cognitive function (Higgins et al. 2004). While speculative, this hypothesis provides a critical impetus for determining the specific group II mGlu subtype responsible for this response and establishing the impact of selectively blocking this receptor on hippocampal function.

Compounds will be initially screened by performing concentration-response curves (CRCs, 10 points, ranging from approximately 30 uM-1 nM at 0.3% final DMSO concentration) to determine the potency and efficacy of novel compounds for mGlu3 in a thallium flux assay measuring coupling of mGlu3 to G Protein-coupled Inwardly Rectifying Potassium (GIRK) Channels (Niswender et al. 2008; Jin et al. 2010; Dhanya et al. 2011). Compounds with IC50 values less than 1 uM will next be evaluated for potency versus mGlu2 in a thallium flux CRC assay. Compounds demonstrating at least 20-fold selectivity for mGlu3 versus mGlu2 will then be evaluated for the mechanism of mGlu3 antagonism through Schild functional thallium flux assays at mGlu3 to determine if the compounds are antagonizing mGlu3 in a competitive or noncompetitive manner. For noncompetetive compounds passing the above steps, we will pursue follow-up studies to determine their selectivity for mGlu3 relative to other mGlu subtypes. Our goal for this project would be to generate an mGlu3-selective compound, with an IC50 under 1 uM, and with reasonable solubility in a solvent generally useful for in vitro experimentation (i.e., DMSO). The ultimate goal of this project from the PI's perspective is to generate compounds that would be useful proof-of-concept molecules to determine the mGlu subtype involved in glial-neuronal communication in the hippocampus.

REFERERENCES
1. Anwyl, R. (1999). "Metabotropic glutamate receptors: electrophysiological properties and role in plasticity." Brain Res Brain Res Rev 29(1): 83-120.
2. Brown, T. A. and A. M. Zador (1990). Hippocampus. The Synaptic Organization of the Brain. S. GM. New York, Oxford UP: 346-388.
3. Coutinho, V. and T. Knopfel (2002). "Metabotropic glutamate receptors: electrical and chemical signaling properties." Neuroscientist 8(6): 551-61.
4. Dhanya, R. P., S. Sidique, et al. (2011). "Design and synthesis of an orally active metabotropic glutamate receptor subtype-2 (mGluR2) positive allosteric modulator (PAM) that decreases cocaine self-administration in rats." J Med Chem 54(1): 342-53.
5. Doherty, J. J., S. Alagarsamy, et al. (2004). "Metabotropic glutamate receptors modulate feedback inhibition in a developmentally regulated manner in rat dentate gyrus." J Physiol 561(Pt 2): 395-401.
6. Fitzjohn, S. M., A. E. Kingston, et al. (1999). "DHPG-induced LTD in area CA1 of juvenile rat hippocampus; characterisation and sensitivity to novel mGlu receptor antagonists." Neuropharmacology 38(10): 1577-83.
7. Gereau, R. W. t. and P. J. Conn (1995b). "Roles of specific metabotropic glutamate receptor subtypes in regulation of hippocampal CA1 pyramidal cell excitability." J Neurophysiol 74(1): 122-9.
8. Jin, X., S. Semenova, et al. (2010). "The mGluR2 positive allosteric modulator BINA decreases cocaine self-administration and cue-induced cocaine-seeking and counteracts cocaine-induced enhancement of brain reward function in rats." Neuropsychopharmacology 35(10): 2021-36.
9. Macek, T. A., D. G. Winder, et al. (1996). "Differential involvement of group II and group III mGluRs as autoreceptors at lateral and medial perforant path synapses." J Neurophysiol 76(6): 3798-806.
10. Moldrich, R. X., K. Aprico, et al. (2002). "Astrocyte mGlu(2/3)-mediated cAMP potentiation is calcium sensitive: studies in murine neuronal and astrocyte cultures." Neuropharmacology 43(2): 189-203.
11. Nicholls, R. E., X. L. Zhang, et al. (2006). "mGluR2 acts through inhibitory Galpha subunits to regulate transmission and long-term plasticity at hippocampal mossy fiber-CA3 synapses." Proc Natl Acad Sci U S A 103(16): 6380-5.
12. Niswender, C. M., K. A. Johnson, et al. (2008). "A novel assay of Gi/o-linked G protein-coupled receptor coupling to potassium channels provides new insights into the pharmacology of the group III metabotropic glutamate receptors." Mol Pharmacol 73(4): 1213-24.
13. Rodriguez, A. L., M. D. Grier, et al. (2010). "Discovery of novel allosteric modulators of metabotropic glutamate receptor subtype 5 reveals chemical and functional diversity and in vivo activity in rat behavioral models of anxiolytic and antipsychotic activity." Mol Pharmacol 78(6): 1105-23.
14. Winder, D. G., P. S. Ritch, et al. (1996). "Novel glial-neuronal signalling by coactivation of metabotropic glutamate and beta-adrenergic receptors in rat hippocampus." J Physiol 494 ( Pt 3): 743-55.
Panel Information
mGlu3 Schild Assay - The effect of multiple concentrations of test compound on a glutamate dose response
PID§NameSubstancePanel TargetsDescription
ActiveInactive
13_uM_Test_CompoundGlutamate dose response with 3 uM Test Compound
210_uM_Test_CompoundGlutamate dose response with 10 uM Test Compound
330_uM_Test_CompoundGlutamate dose response with 30 uM Test Compound

§ Panel component ID.
Protocol
mGlu3 GIRK Schild
Culture of the rat mGlu3 GIRK cell line
HEK/GIRK cells stably expressing the M4 muscarinic receptor and rat mGlu3 were grown in 45% Dulbecco's Modified Eagle Media (DMEM), 45% Ham's F12, 10% fetal bovine serum (FBS), 100 units/ml penicillin/streptomycin, 20 mM HEPES (pH 7.3), 1 mM sodium pyruvate, 2 mM glutamine, 600 ng/ml puromycin dihydrochloride (Sigma-Aldrich) and 700 ug/ml G418 (Mediatech, Inc., Herndon, VA). (Growth Media). Cells for experiments were generally maintained for approximately 15-20 passages. All cell culture reagents were purchased from Invitrogen Corp. (Carlsbad, CA) unless otherwise noted.
Schild Protocol
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% 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% CO2. The following day, plated cells had their medium exchanged to Assay Buffer (Hanks Balanced Salt Solution (Invitrogen) containing 20 mM HEPES pH 7.3) using an ELX405 uplate washer (BioTek), leaving 20 uL/well, followed by addition of with 20 uL of 330 nM FluoZin-2 AM (Invitrogen, Carlsbad, CA) prepared as a 2.85 mM stock in DMSO and mixed in a 1:1 ratio with 10 percent (w/v) pluronic acid F-127 and diluted in Assay Buffer for 1 hour at room temperature. The dye was then exchanged to Assay Buffer using an ELX405, leaving 20 uL/well and the plates were incubated at room temperature for 10 min prior to assay. Test compounds were prepared in DMSO, and transferred to daughter plates using an Echo acoustic plate reformatter (Labcyte, Sunnyvale, CA), and diluted into Assay Buffer to generate a 2x stock. Glutamate concentration-responses were 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. Thallium flux was measured using the Functional Drug Screening System 6000 or 7000 (FDSS6000 or FDSS7000, Hamamatsu, Japan). Baseline readings were taken (10 images at 1 Hz, excitation, 470+/-20 nm, emission, 540+/-30 nm) and then 20 uL/well of test compounds or vehicle were added using the FDSS's integrated pipettor. Approximately 2.5 minutes later 10 uL of Thallium Buffer +/- a concentration-response of glutamate was added. After the addition of glutamate, data were collected for an approximately 3 additional 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. The log of the dose ratio will be calculated and linear regression performed to determine if responses are consistent with competitive (orthosteric) or noncompetitive (allosteric) interaction.
Comment
Compounds showing a Schild slope significantly different from unity (1.0) will be assigned 'Mechanism' = 'Allosteric'.
Result Definitions
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TIDNameDescriptionPID§Panel TargetsHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Fold_Shift_1The EC50 of the glutamate response in the presence of compound divided by the EC50 of the glutamate response in the presence of vehicle1Float
2Fold_Shift_2The EC50 of the glutamate response in the presence of compound divided by the EC50 of the glutamate response in the presence of vehicle2Float
3Fold_Shift_3The EC50 of the glutamate response in the presence of compound divided by the EC50 of the glutamate response in the presence of vehicle3Float
4Compound_Concentration_uM_1Concentration of compound tested in assay in micromolar1FloatμM
5Compound_Concentration_uM_2Concentration of compound tested in assay in micromolar2FloatμM
6Compound_Concentration_uM_3Concentration of compound tested in assay in micromolar3FloatμM
7EC50_uM_1EC50 value in micromolar1FloatμM
8EC50_uM_2EC50 value in micromolar2FloatμM
9EC50_uM_3EC50 value in micromolar3FloatμM
10%_Glu_Max_1Percent Maximum Glutamate Response1Float%
11%_Glu_Max_2Percent Maximum Glutamate Response2Float%
12%_Glu_Max_3Percent Maximum Glutamate Response3Float%
13Category_1Whether the compound is categorized as inactive, a potentiator, or an antagonist1String
14Category_2Whether the compound is categorized as inactive, a potentiator, or an antagonist2String
15Category_3Whether the compound is categorized as inactive, a potentiator, or an antagonist3String
16Mechanism_1Whether the mechanism of action is orthosteric or allosteric1String
17Mechanism_2Whether the mechanism of action is orthosteric or allosteric2String
18Mechanism_3Whether the mechanism of action is orthosteric or allosteric3String
19Value_at_100_uM_R1_1 (100μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
20Value_at_30_uM_R1_1 (30μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
21Value_at_10_uM_R1_1 (10μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
22Value_at_3_uM_R1_1 (3μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
23Value_at_1_uM_R1_1 (1μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
24Value_at_0.3_uM_R1_1 (0.3μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
25Value_at_0.1_uM_R1_1 (0.1μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
26Value_at_0.03_uM_R1_1 (0.03μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
27Value_at_0.01_uM_R1_1 (0.01μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
28Value_at_0.003_uM_R1_1 (0.003μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
29Value_at_0.001_uM_R1_1 (0.001μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
30Value_at_0.0003_uM_R1_1 (0.0003μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
31Value_at_100_uM_R2_1 (100μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
32Value_at_30_uM_R2_1 (30μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
33Value_at_10_uM_R2_1 (10μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
34Value_at_3_uM_R2_1 (3μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
35Value_at_1_uM_R2_1 (1μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
36Value_at_0.3_uM_R2_1 (0.3μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
37Value_at_0.1_uM_R2_1 (0.1μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
38Value_at_0.03_uM_R2_1 (0.03μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
39Value_at_0.01_uM_R2_1 (0.01μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
40Value_at_0.003_uM_R2_1 (0.003μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
41Value_at_0.001_uM_R2_1 (0.001μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
42Value_at_0.0003_uM_R2_1 (0.0003μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).1Float
43Value_at_100_uM_R1_2 (100μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
44Value_at_30_uM_R1_2 (30μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
45Value_at_10_uM_R1_2 (10μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
46Value_at_3_uM_R1_2 (3μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
47Value_at_1_uM_R1_2 (1μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
48Value_at_0.3_uM_R1_2 (0.3μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
49Value_at_0.1_uM_R1_2 (0.1μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
50Value_at_0.03_uM_R1_2 (0.03μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
51Value_at_0.01_uM_R1_2 (0.01μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
52Value_at_0.003_uM_R1_2 (0.003μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
53Value_at_0.001_uM_R1_2 (0.001μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
54Value_at_0.0003_uM_R1_2 (0.0003μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
55Value_at_100_uM_R2_2 (100μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
56Value_at_30_uM_R2_2 (30μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
57Value_at_10_uM_R2_2 (10μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
58Value_at_3_uM_R2_2 (3μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
59Value_at_1_uM_R2_2 (1μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
60Value_at_0.3_uM_R2_2 (0.3μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
61Value_at_0.1_uM_R2_2 (0.1μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
62Value_at_0.03_uM_R2_2 (0.03μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
63Value_at_0.01_uM_R2_2 (0.01μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
64Value_at_0.003_uM_R2_2 (0.003μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
65Value_at_0.001_uM_R2_2 (0.001μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
66Value_at_0.0003_uM_R2_2 (0.0003μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).2Float
67Value_at_100_uM_R1_3 (100μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
68Value_at_30_uM_R1_3 (30μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
69Value_at_10_uM_R1_3 (10μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
70Value_at_3_uM_R1_3 (3μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
71Value_at_1_uM_R1_3 (1μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
72Value_at_0.3_uM_R1_3 (0.3μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
73Value_at_0.1_uM_R1_3 (0.1μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
74Value_at_0.03_uM_R1_3 (0.03μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
75Value_at_0.01_uM_R1_3 (0.01μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
76Value_at_0.003_uM_R1_3 (0.003μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
77Value_at_0.001_uM_R1_3 (0.001μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
78Value_at_0.0003_uM_R1_3 (0.0003μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
79Value_at_100_uM_R2_3 (100μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
80Value_at_30_uM_R2_3 (30μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
81Value_at_10_uM_R2_3 (10μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
82Value_at_3_uM_R2_3 (3μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
83Value_at_1_uM_R2_3 (1μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
84Value_at_0.3_uM_R2_3 (0.3μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
85Value_at_0.1_uM_R2_3 (0.1μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
86Value_at_0.03_uM_R2_3 (0.03μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
87Value_at_0.01_uM_R2_3 (0.01μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
88Value_at_0.003_uM_R2_3 (0.003μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
89Value_at_0.001_uM_R2_3 (0.001μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float
90Value_at_0.0003_uM_R2_3 (0.0003μM**)Normalized fluorescence corrected for baseline and expressed as percent of measured ECmax value (see Protocol).3Float

** Test Concentration. § Panel component ID.
Additional Information
Grant Number: R01 NS031373

Classification
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