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View Project devellis-5U24NS043562-02
Project Summary
Status: Public  
Publications: 1 Published
1 In Preparation
 		 
Project Detail Data Detail
Platform: Affymetrix MIAME Areas Compliance
Species: Rat Array Design Detail false
Organ/Tissue Type: brain Experiment Detail true
Organ Region: cerebral cortex Sample Detail false
Cell Type: primary neuronal cultures Hybridization Detail false
Study Type: time_series_design Measurement Detail false
Disease/Condition: brain development
Replicates: 0
Expected Samples: 112 
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Available Actions
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Investigator Contact Detail
Name Dr. Jean Devellis
Institution: Mental Retardation Research Center, UCLA
Street Address: 635 Charles E Young Dr South
NRB-375D
City, State/Province: LOS ANGELES , CA
Zip/Postal Code: 90095
Country: United States
Work Phone: 310 825 9395
Fax: 310 206 5061
E-mail: jdevellis@mednet.ucla.edu
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Proposal Detail
Grant: 5U24NS043562-02
Status: Public
Service Type: Start to Finish Profiling
IACUC: 1992-034-33
IACUC date: 2005-01-07
Study Relevance:
Nicotinic receptors (nAChR) are widely distributed in the adult brain where they mediate excitatory postsynaptic transmission and presynaptic release of other neurotransmitters. Functional nicotinic receptors are also present in the embryonic brain. Several studies have suggested a role for nAChRs in cognitive functions (learning and memory) and cellular events such as degeneration and survival. In vitro, activation of nAChR can differentially modulate neuritic outgrowth, and enhance early gene expression prior to neuronal differentiation. Furthermore, several reports have shown that nicotine can prevent cell death induced by activation of glutamatergic receptors, NMDA-subtype. Little is known of these events in the cerebral cortex. Knowledge of the genes activated by exposure of cortical neurons to nicotine or to an excitotoxic agent such as the glutamatergic agonist NMDA could potentially lead to new strategies for adult brain protection and repair.
Hypothesis:
We hypothesize that the neuroprotective effect of nAChRs is mediated by modulation of the changes in gene expressions elicited by NMDA.
Specific Aim:
The effect of nicotine and the cembranoid 4R, a novel nicotinic receptor non competitive antagonist, will be analyzed in primary cultures of cortical neurons, in the presence or the absence of NMDA, to identify which genes affected by NMDA receptor activation are modulated nicotine or 4R.
Experimental Procedure and Design:
In order to detect early changes in gene expression, primary culture of cortical neurons prepared from 16 day old Sprague Dawley rat embryos at 11 days in culture will be treated for different times (10, 30, 60, 180, 360, min) with 1 mM NMDA and then harvested. In a second set of experiments, neuronal cultures will be treated with NMDA for 10, 30 and 60 min then the drug will be washed out and the cells will be harvested 24 h later. It was reported that 8-24 h pretreatment of neurons with 10 micromolar nicotine is neuroprotective, therefore we will treat separate cultures with either 10 micromolar nicotine or 2 micromolar 4R for 18 h and then with NMDA for 30 min or 1 h. The cells will be either harvested soon after NMDA treatment or the drugs will be washed out and the cells harvested 24 h later. We will also examine sample treated only with either 4R or nicotine for 15 min or 18 h in order to determine which genes are differentially regulated by these drugs. These experiments will allow us to detect any change in gene expression elicited soon by each treatment or with some delay. The samples in Trizol will be kept at -70 until all the 3 or 5 replicates are harvested and then the RNA will be purified. RNA from 3-5 independent experiments for each different protocol or treatment together with controls will be sent to the facility.
Experimental Factors:
Conditions that are tested in the experiment. At least one is required. Experimental factors are the independent variables in the experiment.
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Factor Name Description Factor Category
specific activation or blockage of nicotinic neurotransmission Nicotine 10uM or 4R 2uM will be appli... timepoint
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Project Samples  This section lists the samples that are associated with this project. Individual sample details can be viewed by clicking on the View Sample icon to the right of the sample. If samples are selectable for analysis or for addition to a virtual
Samples associated with this project.
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Name Description Bio-Source Extracts  
9143 Control (5/7) (Experiment N... BioSource17 1
9138 Control (6/24) (Experiment ... BioSource17 1
9148 Control 7/8 (Experiment NMD... BioSource17 1
9153 Control (7/8RC) (Experiment... BioSource17 1
9144 NMDA 10 (5/7) (Experiment N... BioSource17 2
9139 NMDA 10 (6/24) (Experiment ... BioSource17 1
9149 NMDA 10 (7/8) (Experiment N... BioSource17 1
9154 NMDA 10 (7/8 RC) (Experimen... BioSource17 2
9146 NMDA 1H (5/7) (Experiment N... BioSource17 1
9141 NMDA 1H 6/24 (Experiment NM... BioSource17 1
9151 NMDA 1H 7/8 (Experiment NMD... BioSource17 1
9156 NMDA 1h 7/8 RC (Experiment ... BioSource17 2
9145 NMDA 30 5/7(Experiment NMDA... BioSource17 1
9140 NMDA 30 6/24(Experiment NMD... BioSource17 1
9150 NMDA 30 7/8 (Experiment NMD... BioSource17 2
9155 NMDA 30 7/8RC (Experiment N... BioSource17 1
9602 Control (Experiment NMDA2-R... BioSource17 1
9605 NMDA 1h (Experiment NMDA2-R... BioSource17 1
9604 NMDA 30 (Experiment NMDA2-R... BioSource17 1
9603 NMDA 10 (Experiment NMDA2-R... BioSource17 1
13397 NMDA 10 (7/8) (Experiment N... BioSource17 1
13402 NMDA 10 (7/8 RC) (Experimen... BioSource17 1
13403 NMDA 1h 7/8 RC (Experiment ... BioSource17 1
13399 NMDA 30 7/8 (Experiment NMD... BioSource17 1
13278 Control 8/23/02 (Experiment... BioSource17 1
13279 Control 8/30/02 (Experiment... BioSource17 1
13280 Control 9/06/02 (Experiment... BioSource17 1
13281 nico 18h 8/23/02 (Experimen... BioSource17 1
13282 nico 18h 8/30/02 (Experimen... BioSource17 1
13283 nico 18h 9/06/02 (Experimen... BioSource17 1
13284 nmda 1h 8/23/02 (Experiment... BioSource17 1
13285 nmda 1h 8/30/02 (Experiment... BioSource17 1
13286 nmda 1h 9/06/02 (Experiment... BioSource17 1
13287 nico 18h + nmda 1h 8/23/02 ... BioSource17 1
13288 nico 18h + nmda 1h 8/30/02 ... BioSource17 1
13289 nico 18h + nmda 1h 9/06/02 ... BioSource17 1
14987 Control 1 (Experiment Neuro... BioSource17 1
14988 Control 2 (Experiment Neuro... BioSource17 1
14989 Control 3 (Experiment Neuro... BioSource17 1
14990 Control 4 (Experiment Neuro... BioSource17 1
14991 Control 5 (Experiment Neuro... BioSource17 1
14992 nic 1 (Experiment Neuroprot... BioSource17 1
14993 nic 2 (Experiment Neuroprot... BioSource17 1
14994 nic 3 (Experiment Neuroprot... BioSource17 1
14995 nic 4 (Experiment Neuroprot... BioSource17 1
14996 nic 5 (Experiment Neuroprot... BioSource17 1
14997 nmda1 (Experiment Neuropro... BioSource17 1
14998 nmda2 (Experiment Neuropro... BioSource17 1
14999 nmda3 (Experiment Neuroprot... BioSource17 1
15000 nmda4 (Experiment Neuropro... BioSource17 1
15001 nmda5 (Experiment Neuropro... BioSource17 1
15002 nic + nmda1 (Experiment Ne... BioSource17 1
15003 nic + nmda2 (Experiment Ne... BioSource17 1
15004 nic + nmda3 (Experiment Neu... BioSource17 1
15005 nic + nmda4 (Experiment Neu... BioSource17 1
15006 nic + nmda5 (Experiment Neu... BioSource17 1
19224 CO (Experiment shortNMDA: R... BioSource17 1
19231 CO (Experiment shortNMDA: R... BioSource17 1
19238 CO (Experiment shortNMDA: R... BioSource17 1
19225 NMDA10 (Experiment shortNMD... BioSource17 1
19232 NMDA10 (Experiment shortNMD... BioSource17 1
19239 NMDA10 (Experiment shortNMD... BioSource17 1
19226 NMDA30 (Experiment shortNMD... BioSource17 1
19233 NMDA30 (Experiment shortNMD... BioSource17 1
19240 NMDA30 (Experiment shortNMD... BioSource17 1
19227 NMDA60 (Experiment shortNMD... BioSource17 1
19234 NMDA60 (Experiment shortNMD... BioSource17 1
19241 NMDA60 (Experiment shortNMD... BioSource17 1
19228 NMDA180 (Experiment shortNM... BioSource17 1
19235 NMDA180 (Experiment shortNM... BioSource17 1
19242 NMDA180 (Experiment shortNM... BioSource17 1
19229 NMDA360 (Experiment shortNM... BioSource17 1
19236 NMDA360 (Experiment shortNM... BioSource17 1
19243 NMDA360 (Experiment shortNM... BioSource17 1
19285 CO (Experiment 4Rlong: Run ... BioSource17 1
19286 CO (Experiment 4Rlong: Run ... BioSource17 1
19287 CO (Experiment 4Rlong: Run ... BioSource17 1
19288 4R-3 1/29 (Experiment 4Rlon... BioSource17 1
19289 4R-4 2/5 (Experiment 4Rlong... BioSource17 1
19290 4R-5 2/5CG (Experiment 4Rlo... BioSource17 1
19871 CO 1 (Experiment 4R18h: Run... BioSource17 1
19872 CO 2 (Experiment 4R18h: Run... BioSource17 1
19873 CO 3 (Experiment 4R18h: Run... BioSource17 1
19874 4R 1 (Experiment 4R18h: Run... BioSource17 1
19875 4R 2 (Experiment 4R18h: Run... BioSource17 1
19876 4R 3 (Experiment 4R18h: Run... BioSource17 1
21825 Control 1 (Experiment Neuro... BioSource17 1
21826 Control 2 (Experiment Neuro... BioSource17 1
21827 Control 3 (Experiment Neuro... BioSource17 1
21828 Control 4 (Experiment Neuro... BioSource17 1
21829 Control 5 (Experiment Neuro... BioSource17 1
21830 nic 1 (Experiment Neuroprot... BioSource17 1
21831 nic 2 (Experiment Neuroprot... BioSource17 1
21832 nic 3 (Experiment Neuroprot... BioSource17 1
21833 nic 4 (Experiment Neuroprot... BioSource17 1
21834 nic 5 (Experiment Neuroprot... BioSource17 1
21835 nmda1 (Experiment Neuropro... BioSource17 1
21836 nmda2 (Experiment Neuropro... BioSource17 1
21837 nmda3 (Experiment Neuroprot... BioSource17 1
21838 nmda4 (Experiment Neuropro... BioSource17 1
21839 nmda5 (Experiment Neuropro... BioSource17 1
21840 nic + nmda1 (Experiment Ne... BioSource17 1
21841 nic + nmda2 (Experiment Ne... BioSource17 1
21842 nic + nmda3 (Experiment Neu... BioSource17 1
21843 nic + nmda4 (Experiment Neu... BioSource17 1
21844 nic + nmda5 (Experiment Neu... BioSource17 1
23147 Co 1 (Experiment Nic 15 - R... BioSource17 1
23148 Co 2 (Experiment Nic 15 - R... BioSource17 1
23149 Co 3 (Experiment Nic 15 - R... BioSource17 1
23150 Nic 1 (Experiment Nic 15 -... BioSource17 1
23151 Nic 2 (Experiment Nic 15 - ... BioSource17 1
23152 Nic 3 (Experiment Nic 15 - ... BioSource17 1
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Project Hybridizations 

Action Button Key
View Hybridization View Hybridization  
Name Array Labeled Extract Hybridization Protocol  
RN_U34_1_hyb RN_U34_1 9149_e2_le1
RN_U34_2_hyb RN_U34_2 Co 5/7_le1
RN_U34_3_hyb RN_U34_3 Co 6/24_le1
RN_U34_4_hyb RN_U34_4 Co 7/8_le1
RN_U34_5_hyb RN_U34_5 Co 7/8 RC_le1
RN_U34_6_hyb RN_U34_6 NMDA10 5/7_le1
RN_U34_7_hyb RN_U34_7 NMDA10 6/24_le1
RN_U34_8_hyb RN_U34_8 NMDA10 7/8_le1
RN_U34_9_hyb RN_U34_9 NMDA10 7/8RC_le1
RN_U34_10_hyb RN_U34_10 NMDA1H 5/7_le1
RN_U34_11_hyb RN_U34_11 NMDA1H 6/24_le1
RN_U34_12_hyb RN_U34_12 NMDA1H 7/8_le1
RN_U34_13_hyb RN_U34_13 NMDA1H 7/8RC_le1
RN_U34_14_hyb RN_U34_14 NMDA30 5/7_le1
RN_U34_15_hyb RN_U34_15 NMDA30 6/24_le1
RN_U34_16_hyb RN_U34_16 NMDA30 7/8_le1
RN_U34_17_hyb RN_U34_17 NMDA30 7/8RC_le1
RN_U34_18_hyb RN_U34_18 Control_le1
RN_U34_19_hyb RN_U34_19 NMDA 1mM 1HOUR_le1
RN_U34_20_hyb RN_U34_20 NMDA 1mM 30'_le1
RN_U34_21_hyb RN_U34_21 NMDA 1mM 10'_le1
RN_U34_22_hyb RN_U34_22 9154_e3_le1
RN_U34_23_hyb RN_U34_23 9156_e3_le1
RN_U34_24_hyb RN_U34_24 9150_e3_le1
RN_U34_25_hyb RN_U34_25 Control 8/23_le1
RN_U34_26_hyb RN_U34_26 Control 8/30_le1
RN_U34_27_hyb RN_U34_27 Control 9/06_le1
RN_U34_28_hyb RN_U34_28 Nico 8/23_le1
RN_U34_29_hyb RN_U34_29 Nico 8/30_le1
RN_U34_30_hyb RN_U34_30 Nico 9/06_le1
RN_U34_31_hyb RN_U34_31 NMDA 8/23_le1
RN_U34_32_hyb RN_U34_32 NMDA 8/30_le1
RN_U34_33_hyb RN_U34_33 NMDA 9/06_le1
RN_U34_34_hyb RN_U34_34 Nico + NMDA 8/23_le1
RN_U34_35_hyb RN_U34_35 Nico + NMDA 8/30_le1
RN_U34_36_hyb RN_U34_36 Nico + NMDA 9/06_le1
RN_U34_37_hyb RN_U34_37 Control 1_le1
RN_U34_38_hyb RN_U34_38 Control 2_le1
RN_U34_39_hyb RN_U34_39 Control 3_le1
RN_U34_40_hyb RN_U34_40 Control 4_le1
RN_U34_41_hyb RN_U34_41 Control 5_le1
RN_U34_42_hyb RN_U34_42 Nic 1_le1
RN_U34_43_hyb RN_U34_43 Nic 2_le1
RN_U34_44_hyb RN_U34_44 Nic 3_le1
RN_U34_45_hyb RN_U34_45 Nic 4_le1
RN_U34_46_hyb RN_U34_46 Nic 5_le1
RN_U34_47_hyb RN_U34_47 NMDA 1_le1
RN_U34_48_hyb RN_U34_48 NMDA 2_le1
RN_U34_49_hyb RN_U34_49 NMDA 3_le1
RN_U34_50_hyb RN_U34_50 NMDA 4_le1
RN_U34_51_hyb RN_U34_51 NMDA 5_le1
RN_U34_52_hyb RN_U34_52 NIC + NMDA 1_le1
RN_U34_53_hyb RN_U34_53 NIC + NMDA 2_le1
RN_U34_54_hyb RN_U34_54 NIC + NMDA 3_le1
RN_U34_55_hyb RN_U34_55 NIC + NMDA 4_le1
RN_U34_56_hyb RN_U34_56 NIC + NMDA 5_le1
RN_U34_57_hyb RN_U34_57 CO2_le1
RN_U34_58_hyb RN_U34_58 CO3_le1
RN_U34_59_hyb RN_U34_59 CO4_le1
RN_U34_60_hyb RN_U34_60 NMDA 10 2_le1
RN_U34_61_hyb RN_U34_61 NMDA 10 3_le1
RN_U34_62_hyb RN_U34_62 NMDA 10 4_le1
RN_U34_63_hyb RN_U34_63 NMDA 30 2_le1
RN_U34_64_hyb RN_U34_64 NMDA 30 3_le1
RN_U34_65_hyb RN_U34_65 NMDA 30 4_le1
RN_U34_66_hyb RN_U34_66 NMDA 60 2_le1
RN_U34_67_hyb RN_U34_67 NMDA 60 3_le1
RN_U34_68_hyb RN_U34_68 NMDA 60 4_le1
RN_U34_69_hyb RN_U34_69 NMDA 180 2_le1
RN_U34_70_hyb RN_U34_70 NMDA 180 3_le1
RN_U34_71_hyb RN_U34_71 NMDA 180 4_le1
RN_U34_72_hyb RN_U34_72 NMDA 360 2_le1
RN_U34_73_hyb RN_U34_73 NMDA 360 3_le1
RN_U34_74_hyb RN_U34_74 NMDA 369 4_le1
RN_U34_75_hyb RN_U34_75 Co3_le1
RN_U34_76_hyb RN_U34_76 Co4 (2/5)_le1
RN_U34_77_hyb RN_U34_77 Co4 (2/5CG)_le1
RN_U34_78_hyb RN_U34_78 4R-3(1/29)_le1
RN_U34_79_hyb RN_U34_79 4R-4(2/5)_le1
RN_U34_80_hyb RN_U34_80 4R-4(2/5CG)_le1
RN_U34_81_hyb RN_U34_81 Co 1 (4/14)_le1
RN_U34_82_hyb RN_U34_82 Co 2 (4/22)_le1
RN_U34_83_hyb RN_U34_83 Co 3 (4/28)_le1
RN_U34_84_hyb RN_U34_84 4R 1 (4/14)_le1
RN_U34_85_hyb RN_U34_85 4R 2 (4/22)_le1
RN_U34_86_hyb RN_U34_86 4R 3 (4/28)_le1
RN_U34_87_hyb RN_U34_87 14987_e2_le1
RN_U34_88_hyb RN_U34_88 14988_e2_le1
RN_U34_89_hyb RN_U34_89 14989_e2_le1
RN_U34_90_hyb RN_U34_90 14990_e2_le1
RN_U34_91_hyb RN_U34_91 14991_e2_le1
RN_U34_92_hyb RN_U34_92 14992_e2_le1
RN_U34_93_hyb RN_U34_93 14993_e2_le1
RN_U34_94_hyb RN_U34_94 14994_e2_le1
RN_U34_95_hyb RN_U34_95 14995_e2_le1
RN_U34_96_hyb RN_U34_96 14996_e2_le1
RN_U34_97_hyb RN_U34_97 14997_e2_le1
RN_U34_98_hyb RN_U34_98 14998_e2_le1
RN_U34_99_hyb RN_U34_99 14999_e2_le1
RN_U34_100_hyb RN_U34_100 15000_e2_le1
RN_U34_101_hyb RN_U34_101 15001_e2_le1
RN_U34_102_hyb RN_U34_102 15002_e2_le1
RN_U34_103_hyb RN_U34_103 15003_e2_le1
RN_U34_104_hyb RN_U34_104 15004_e2_le1
RN_U34_105_hyb RN_U34_105 15005_e2_le1
RN_U34_106_hyb RN_U34_106 15006_e2_le1
RN_U34_107_hyb RN_U34_107 Co1_le1
RN_U34_108_hyb RN_U34_108 Co4_le1
RN_U34_109_hyb RN_U34_109 Co5_le1
RN_U34_110_hyb RN_U34_110 Nic1_le1
RN_U34_111_hyb RN_U34_111 Nic4_le1
RN_U34_112_hyb RN_U34_112 Nic5_le1
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