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Status |
Public on Jun 06, 2006 |
Title |
Thrivikraman-5P50MH058922-050002 |
Organism |
Rattus norvegicus |
Experiment type |
Expression profiling by array
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Summary |
It is suggested that the stress induced activation of the HPA axis and associated increases in plasma ACTH and glucocorticoids (corticosterone, B in the rat) are contributing factors in the development of many psychopathologies. However, the circadian fluctuation of the HPA axis activity (characterized by a zenith in the plasma ACTH and B, before the beginning of the active awake state and the nadir during the inactive sleeping period) occurs throughout the life and is important for normal physiological and behavioral functioning. A blunting or enhancement of this rhythm through changes in the trough or in the peak is a characteristic feature of many pathological states. At the CNS level, the circadian fluctuation of the HPA axis activity is associated with changes in the expression profile of structural, functional, and immediate early genes. However, little is known about the specific role of B in the modulation of the circadian pattern of gene regulation in the CNS. Thus, during the circadian cycle B is a major factor, which could influence the expression profile of a large family of genes. Accordingly, the microarray technology, because of its characteristic feature that facilitates the identification of the expression profiles of a wide range of genes simultaneously, is an ideal technique. Because the B status of animals can be easily manipulated by adrenalectomy (ADX) surgery with or without B replacement, this model will be used in these studies. Thus, the proposed studies are intended to characterize, by the use of the microarray technique, the effects of the B milieu on diurnal gene expression profiles in selected rat brain regions strongly influenced by: a.) B status: amygdala, hippocampus, hypothalamus, and septum; and b.) Circadian Activity: dorsal raphe, frontal Cortex and locus coeruleus. The gradient of the B response during the circadian cycle implies that at the beginning of the sleep cycle MR are predominantly occupied while at the end of the sleep cycle, both MR and GR are occupied. Thus, we propose that differing ratios of activation of these nuclear receptors could differentially modulate gene expression and impart temporal characteristics in the expression profile of a large family of genes. Furthermore, we propose that these effects may be region specific, reflecting regional differences in GR and MR distribution, B uptake, and neural activity. B may exert these circadian effects on regional gene expression profiles either directly or indirectly via interneurons. However, little is known about the circadian effect of B on gene expression profiles in different brain regions. Such knowledge is important to identify the specific role of B in circadian differences in CNS function and, potentially, the development of psychopathologies. After arrival at the facility the rats (male Long Evans) will be maintained on 12:12 h LD cycle ('lights-on" at 0700 h) with food and water ad libitum. One week after, they will undergo bilateral ADX under survival anesthetics (day-0). The study will include three groups (n=10); (a) ADX: rats with no adrenals, (b) pADX: ADX rats with subcutaneous 40 percent B pellet (fused cholesterol and B), and (c) SHAM: rats undergoing flank incisions without the removal of the adrenals. After surgery, rats will be housed individually. The ADX rats will receive 0.5 percent saline in addition to regular tap water. On post-surgical day-7, 15 rats (five/group) will be sacrificed by decapitation 2h after 'lights-on", and the remaining 15, 2h after "lights-off". The brains will be removed immediately, frozen on dry ice, and stored at -80 C. The trunk blood will be collected with EDTA and plasma frozen for ACTH and B RIA. The brains from rats with confirmed ADX will be used for free hand dissection of brain regions: frontal cortex, septum, hypothalamus, amygdala, hippocampus, dorsal raphe, and locus coeruleus. Tissue will be stored at -80 C. Total RNA will be extracted from individual brain regions using Qiagen RNeasy Lipid Tissue kit. Initial evaluation of quality and quantity of the RNA will be done by the absorbance method. Frozen samples containing 0.5 ul total RNA or higher will be sent to the NINDS-NIMH Microarray Consortium in dry ice for evaluation of the integrity of 28S and 18S RNA on an Agilent Bioanalyzer. If the integrity of RNA is not good, the experiment will be repeated and new RNA samples will be provided. Replicates of each brain region with 28S/18S ratio of 1.7 and higher will be selected for assay using Affymetrix Rat U34A array. The RNA samples from one region from all the brains will be processed together. A total of 84 chips may be utilized in these studies, however the final number will depend upon integrity of the total RNA. Keywords: other
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Contributor(s) |
Thrivikraman KV |
Citation missing |
Has this study been published? Please login to update or notify GEO. |
Submission date |
May 04, 2006 |
Last update date |
Jul 31, 2017 |
Contact name |
Winnie Liang |
E-mail(s) |
wliang@tgen.org
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Organization name |
Translational Genomics
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Street address |
445 N. Fifth Street
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City |
Phoenix |
State/province |
AZ |
ZIP/Postal code |
85012 |
Country |
USA |
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Platforms (1) |
GPL1355 |
[Rat230_2] Affymetrix Rat Genome 230 2.0 Array |
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Samples (82)
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GSM107895 |
brain, frontal cortex: LabeledExtract2 |
GSM107896 |
brain, frontal cortex: LabeledExtract3 |
GSM107897 |
brain, frontal cortex: pADX AM FC_e1_le1 |
GSM107898 |
brain, frontal cortex: Sham PM FC_e1_le1 |
GSM107899 |
brain, frontal cortex: ADX PM FC_e1_le1 |
GSM107900 |
brain, frontal cortex: pADX PM FC_e1_le1 |
GSM107901 |
brain, frontal cortex: Sham AM S_e1_le1 |
GSM107902 |
brain, frontal cortex: ADX AM S_e1_le1 |
GSM107903 |
brain, frontal cortex: pADX AM S_e1_le1 |
GSM107904 |
brain, frontal cortex: Sham PM S_e1_le1 |
GSM107905 |
brain, frontal cortex: ADX PM S_e1_le1 |
GSM107906 |
brain, frontal cortex: pADX PM S_e1_le1 |
GSM107907 |
brain, hippocampi, left: Sham AM Hypo_e1_le1 |
GSM107908 |
brain, hippocampi, left: ADX AM Hypo_e1_le1 |
GSM107909 |
brain, hippocampi, left: pADX AM Hypo_e1_le1 |
GSM107910 |
brain, hippocampi, left: Sham PM Hypo_e1_le1 |
GSM107911 |
brain, hippocampi, left: ADX PM Hypo_e1_le1 |
GSM107912 |
brain, hippocampi, left: pADX PM Hypo_e1_le1 |
GSM107913 |
brain, amygdala: Sham AM A_e1_le1 |
GSM107914 |
brain, amygdala: ADX AM A_e1_le1 |
GSM107915 |
brain, amygdala: pADX AM A_e1_le1 |
GSM107916 |
brain, amygdala: Sham PM A_e1_le1 |
GSM107917 |
brain, amygdala: ADX PM A_e1_le1 |
GSM107918 |
brain, amygdala: pADX PM A_e1_le1 |
GSM107919 |
brain, hippocampus: Sham AM Hip_e1_le1 |
GSM107920 |
brain, hippocampus: ADX AM Hip_e1_le1 |
GSM107921 |
brain, hippocampus: pADX PM Hip_e1_le1 |
GSM107922 |
brain, hippocampus: Sham PM Hip_e1_le1 |
GSM107923 |
brain, hippocampus: ADX PM Hip_e1_le1 |
GSM107925 |
brain, frontal cortex: Sham AM DR_e1_le1 |
GSM107926 |
brain, frontal cortex: ADX AM DR_e1_le1 |
GSM107927 |
brain, frontal cortex: pADX AM DR_e1_le1 |
GSM107928 |
brain, frontal cortex: Sham PM DR_e1_le1 |
GSM107929 |
brain, frontal cortex: ADX PM DR_e1_le1 |
GSM107930 |
brain, frontal cortex: pADX PM DR_e1_le1 |
GSM107931 |
brain, frontal cortex: Sham AM LC_e1_le1 |
GSM107932 |
brain, frontal cortex: ADX AM LC_e1_le1 |
GSM107933 |
brain, frontal cortex: pADX AM LC_e1_le1 |
GSM107934 |
brain, frontal cortex: Sham PM LC_e1_le1 |
GSM107935 |
brain, frontal cortex: ADX PM LC_e1_le1 |
GSM107936 |
brain, frontal cortex: pADX PM LC_e1_le1 |
GSM107937 |
brain, frontal cortex: Sham AM FC b_e1_le1 |
GSM107938 |
brain, frontal cortex: ADX AM FC b_e1_le1 |
GSM107939 |
brain, frontal cortex: pADX AM FC b_e1_le1 |
GSM107940 |
brain, frontal cortex: Sham PM FC b_e1_le1 |
GSM107941 |
brain, frontal cortex: ADX PM FC b_e1_le1 |
GSM107942 |
brain, frontal cortex: pADX PM FC b_e1_le1 |
GSM107943 |
brain, frontal cortex: Sham AM S b_e1_le1 |
GSM107944 |
brain, frontal cortex: ADX AM S b_e1_le1 |
GSM107945 |
brain, frontal cortex: pADX AM S b_e1_le1 |
GSM107946 |
brain, frontal cortex: Sham PM S b_e1_le1 |
GSM107947 |
brain, frontal cortex: ADX PM S b_e1_le1 |
GSM107948 |
brain, frontal cortex: pADX PM S b_e1_le1 |
GSM107949 |
brain, hippocampi, left: Sham AM Hypo b_e1_le1 |
GSM107950 |
brain, hippocampi, left: ADX AM Hypo b_e1_le1 |
GSM107951 |
brain, hippocampi, left: pADX AM Hypo b_e1_le1 |
GSM107952 |
brain, hippocampi, left: Sham PM Hypo b_e1_le1 |
GSM107953 |
brain, hippocampi, left: ADX PM Hypo b_e1_le1 |
GSM107954 |
brain, hippocampi, left: pADX PM Hypo b_e1_le1 |
GSM107955 |
brain, amygdala: Sham AM A b_e1_le1 |
GSM107956 |
brain, amygdala: ADX AM A b_e1_le1 |
GSM107957 |
brain, amygdala: pADX AM A b_e1_le1 |
GSM107958 |
brain, amygdala: Sham PM A b_e1_le1 |
GSM107959 |
brain, amygdala: ADX PM A b_e1_le1 |
GSM107960 |
brain, amygdala: pADX PM A b_e1_le1 |
GSM107961 |
brain, hippocampus: Sham AM Hip b_e1_le1 |
GSM107962 |
brain, hippocampus: ADX AM Hip b_e1_le1 |
GSM107963 |
brain, hippocampus: pADX PM Hip b_e1_le1 |
GSM107964 |
brain, hippocampus: Sham PM Hip b_e1_le1 |
GSM107965 |
brain, hippocampus: ADX PM Hip b_e1_le1 |
GSM107966 |
brain, frontal cortex: Sham AM DR b_e1_le1 |
GSM107967 |
brain, frontal cortex: ADX AM DR b_e1_le1 |
GSM107968 |
brain, frontal cortex: pADX AM DR b_e1_le1 |
GSM107969 |
brain, frontal cortex: Sham PM DR b_e1_le1 |
GSM107970 |
brain, frontal cortex: ADX PM DR b_e1_le1 |
GSM107971 |
brain, frontal cortex: pADX PM DR b_e1_le1 |
GSM107972 |
brain, frontal cortex: Sham AM LC b_e1_le1 |
GSM107973 |
brain, frontal cortex: ADX AM LC b_e1_le1 |
GSM107974 |
brain, frontal cortex: pADX AM LC b_e1_le1 |
GSM107975 |
brain, frontal cortex: Sham PM LC b_e1_le1 |
GSM107976 |
brain, frontal cortex: ADX PM LC b_e1_le1 |
GSM107977 |
brain, frontal cortex: pADX PM LC b_e1_le1 |
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Relations |
BioProject |
PRJNA95547 |
Supplementary file |
Size |
Download |
File type/resource |
GSE4776_RAW.tar |
193.9 Mb |
(http)(custom) |
TAR (of CEL) |
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