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| Status |
Public on Feb 09, 2007 |
| Title |
Wildtype Mouse Hypothalamus Cerebellum Fed Fast |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
The hypothalamus is a key site for the integration of the response to nutrient deprivation in higher organisms. Transcripts that are dynamically regulated in the hypothalamus by feeding and fasting are likely to play a regulatory role in energy balance. In order to identify hypothalamic genes whose expression is altered by fasting, we compared the transcriptomes of hypothalamic blocks from freely feeding vs overnight fasted mice using whole genome oligonucleotide arrays. In order to exclude changes due to non-specific neuronal stress responses to energy deprivation we simultaneously profiled the cerebellum, a brain site with no known regulatory role in energy balance. The power of this approach was illustrated by the fact that while we confirmed previous observations that the Sulfotransferase family 1A gene is upregulated by fasting in the hypothalamus, we showed that it was similarly upregulated in the cerebellum and is therefore unlikely to be specifically involved in the regulation of energy balance. In contrast, using three different methods of pathway enrichment analysis, multiple genes involved in mitochondrial oxidative phosphorylation were found to be co-ordinately downregulated by fasting in the hypothalamus but not in the cerebellum. These findings suggest that a switch from oxidative to non-oxidative metabolism in the hypothalamus may be an important element of the homeostatic response to nutrient deprivation.
Keywords: hypothalamus, cerebellum, fasting, appetite, obesity
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| Overall design |
Wild-type female SV/129 mice were purchased from Charles River. At onset of the dark phase the evening before the experiment, food was removed from one group of 6 mice; while a further 6 animals were allowed continued access to chow. Both groups had ad libitum access to water. The next morning (16 hours later), mice were sacrificed by cervical dislocation and decapitated. The brains were removed and the hypothalami and cerebellar slices were rapidly dissected and snap frozen on powdered dry ice. Purified RNA from each condition and brain region was pooled resulting in 4 different sample groups (Fed & Fasted; hypothalamus & cerebellum). Double stranded cDNA was generated from 2μg of total RNA from each sample group, converted to cRNA and biotin labelled using the Codelink Expression Assay Reagent Kit (GE Healthcare). 10μg of biotin labelled cRNA was hybridized overnight onto CodeLink™ Mouse Whole Genome Bioarrays (GE Healthcare) according to manufacturer’s protocols. Each RNA sample group was hybridized in triplicate. One Cerebellum Fast chip was discarded because it failed QC.
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| Contributor(s) |
Curtis RK, Tung YL, Piper SJ, Coll AP, O'Rahilly S, Yeo GS |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| Submission date |
Feb 09, 2006 |
| Last update date |
Mar 25, 2013 |
| Contact name |
Keira Curtis |
| Organization name |
University of Cambridge
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| Department |
Clinical Biochemistry
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| Street address |
Box 232, Addenbrooke's Hospital, Hills Road
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| City |
Cambridge |
| ZIP/Postal code |
CB2 2QR |
| Country |
United Kingdom |
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| Platforms (1) |
| GPL2897 |
GE Healthcare/Amersham Biosciences CodeLink™ Mouse Whole Genome Bioarray |
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| Samples (11)
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| Relations |
| BioProject |
PRJNA94863 |
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