|
Status |
Public on Oct 20, 2010 |
Title |
BL6_4h vs. basal replicate 10 |
Sample type |
RNA |
|
|
Channel 1 |
Source name |
brain
|
Organism |
Mus musculus |
Characteristics |
stress: stressed time point: 4h strain: C57BL/6
|
Treatment protocol |
Mice were subjected to forced swimming in a cylinder of 24cm height and a diameter of 11 cm filled with water at 22-25°C for 5 minutes. Afterwards, they were transferred to their own original cage (i.e. single housed again) until decapitation.
|
Growth protocol |
C57BL/6J and DBA/2J male mice with an age of 3-5 months (supplied from Harlan Winkelman, Borchen) were single housed under a 06:00-18:00 light cycle, where food and water were available ad libitum.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA of PVN micropunctured tissue was extracted using the TRIzol reagent (Invitrogen) according to manufacturer´s instruction. RNA qualitiy was analysed by gel electrophoresis.
|
Label |
Cy3
|
Label protocol |
aRNA synthesis and aRNA labelling was performed with the Amino Allyl MessageAmp™ aRNA Kit (Ambion) following the manufacturer’s protocol using 3 µg of total RNA.
|
|
|
Channel 2 |
Source name |
brain
|
Organism |
Mus musculus |
Characteristics |
stress: basal strain: C57BL/6
|
Treatment protocol |
Mice were subjected to forced swimming in a cylinder of 24cm height and a diameter of 11 cm filled with water at 22-25°C for 5 minutes. Afterwards, they were transferred to their own original cage (i.e. single housed again) until decapitation.
|
Growth protocol |
C57BL/6J and DBA/2J male mice with an age of 3-5 months (supplied from Harlan Winkelman, Borchen) were single housed under a 06:00-18:00 light cycle, where food and water were available ad libitum.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA of PVN micropunctured tissue was extracted using the TRIzol reagent (Invitrogen) according to manufacturer´s instruction. RNA qualitiy was analysed by gel electrophoresis.
|
Label |
Cy5
|
Label protocol |
aRNA synthesis and aRNA labelling was performed with the Amino Allyl MessageAmp™ aRNA Kit (Ambion) following the manufacturer’s protocol using 3 µg of total RNA.
|
|
|
|
Hybridization protocol |
The labeled aRNA samples were hybridized on Max-Planck Institute 24 k mouse cDNA arrays (Max-Planck-Institute of Psychiatry, Munich, Germany) as described at Deussing at al., 2207, J Cereb Blood Flow Metab., performing 3 technical replicates for each control/CRH treated dye coupling combination.
|
Scan protocol |
Mircorarrays were scanned on a PerkinElmer Life Sciences ScanArray 4000 laser scanner.
|
Data processing |
The microarray data analysis was accomplished by the fixed circle quantification method using QuantArray (PerkinElmer Life Sciences) and raw data was normalized using a nonlinear regression method (Yang et al., 2002).
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|
|
Submission date |
Mar 15, 2010 |
Last update date |
Oct 20, 2010 |
Contact name |
Benno Pütz |
E-mail(s) |
puetz@mpipsykl.mpg.de
|
Phone |
++49-(0)89-30622-222
|
Fax |
++49-(0)89-30622-601
|
Organization name |
MPI of Psychiatry
|
Department |
Statistical Genetics
|
Street address |
Kraepelinstr. 2
|
City |
München |
ZIP/Postal code |
80804 |
Country |
Germany |
|
|
Platform ID |
GPL7467 |
Series (1) |
GSE20877 |
Novel aspects of the transcriptional stress response in the paraventricular nucleus of the mouse strains DBA/2J and C57BL/6J |
|