|
| Status |
Public on Oct 04, 2014 |
| Title |
Constipated rats -Vehicle |
| Sample type |
RNA |
| |
|
| Source name |
SD rat
|
| Organism |
Rattus norvegicus |
| Characteristics |
tissue: Distal colon
gender: male
age: 8 weeks
strain: SD
treatment: vehicle-treated constipation
|
| Treatment protocol |
Constipation was induced in SD rats by subcutaneous injection of loperamide (4 mg/kg weight) in 0.9% sodium chloride twice (9 AM and 6 PM) a day for 3 days, whereas the non-constipation group was injected with 0.9% sodium chloride alone as described in previous study. At 15 hr after the final treatment of loperamide, the constipation group was further divided into a vehicle-treated constipation group and AEtLP-treated constipation group. They were received a consistent volume of water or 15 uL/g body weight of AEtLP (1,000 mg/kg weight) via oral administration for once at 9 AM. At 24h after the treatment of AEtLP and vehicle, all animals were immediately euthanatized using CO2 gas and tissue samples were acquired and stored in Eppendorf tubes at -70℃ until assaued.
|
| Growth protocol |
All animals were handled at the Pusan National University Laboratory Animal Resources Center accredited by AAALAC International (Accredited Unit Number; 001525) according to the National Institutes of Health guidelines.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
Amplified and labeled cRNA was purified on cRNA Cleanup Module (Agilent Technology) according to the manufacturer’s protocol. Labeled cRNA target was quantified using ND-1000 spectrophotometer (NanoDrop Technologies, Inc., Wilmington, DE).
|
| |
|
| Hybridization protocol |
After checking labeling efficiency, fragmentation of cRNA was performed by adding 10X blocking agent and 25X fragmentation buffer and incubating at 60oC for 30 min. The fragmented cRNA was resuspended with 2X hybridization buffer and directly pipetted onto assembled Agilent’s Canine Oligo Microarray (44K). The arrays hybridized at 65oC for 17 hours using Agilent Hybridization oven (Agilent Technology, USA). The hybridized microarrays were washed as the manufacturer’s washing protocol (Agilent Technology, USA).
|
| Scan protocol |
The hybridized images were scanned using Agilent’s DNA microarray scanner and quantified with Feature Extraction Software (Agilent Technology, Palo Alto, CA).
|
| Data processing |
All data normalization and further analysis were performed using GeneSpringGX 7.3 (Agilent Technology, USA).
|
| |
|
| Submission date |
Oct 03, 2014 |
| Last update date |
Oct 04, 2014 |
| Contact name |
Dae Youn Hwang |
| E-mail(s) |
[email protected]
|
| Phone |
01072279769
|
| Organization name |
Pusan National University
|
| Street address |
50 Cheonghak-ri, Samnangjin-eup
|
| City |
Miryang |
| ZIP/Postal code |
KS011 |
| Country |
South Korea |
| |
|
| Platform ID |
GPL14746 |
| Series (1) |
| GSE62041 |
Characterization of changes in global genes expression in the distal colon of loperamide-induced constipation SD rats in response to the laxative effects of Liriope platyphylla |
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