|
| Status |
Public on Oct 20, 2010 |
| Title |
BL6_8h vs. basal replicate 9 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
brain
|
| Organism |
Mus musculus |
| Characteristics |
stress: stressed
time point: 8h
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).
|
| |
|
| 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 |
|
