|
| Status |
Public on Oct 18, 2017 |
| Title |
Small intestinal mucosa_1st-infection_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
Small intestinal mucosa, 1st-infection group, 6days after infection, replicate0
|
| Organism |
Canis lupus familiaris |
| Characteristics |
tissue: small intestinal mucosa (including submucosa and muscularis externa)
gender: female
age: 8 month old
|
| Treatment protocol |
For the control group, dogs were euthanized and necropsies were performed. The small intestine was cut out, the central part (approximately 15 cm) was washed with cold saline, then quickly frozen with liquid nitrogen. This samples were preserved in liquid nitrogen until use. For the first-infection group, 100,000 protoscoleces were orally administrated to infect E. multilocularis. For the repeated-infection group (n=4), 500,000 protoscoleces was administrated orally at 1st, 2nd and 3rd infection. At the repeated infection, the infection was terminated 35 days postinfection (dpi) by administrating 100 mg of praziquantel (2 tablets of Droncit®, Bayer-Animal Health). At the final infection, 100,000 protoscoleces was administrated by identical method. After six days of infection, the central part of small intestine from dogs infected with E. multilocularis was cut out as above mentioned method and the samples were treated with RNA later according to the manufacture’s instruction. The sample were preserved at -70°C until use.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Tissue samples from infected-dogs were carefully thawed in RNA later at room temprature. Frozen tissue from normal dogs were treated with RNAlater-ICE according to the manufacture’s instruction. Small intestinal mucosa was scraped with micro spatula on a plastic culture dishes and total RNA was extracted from tissue sample with RNeasy Mini Kit (Qiagen) according to the manufacture’s instruction. Concentration and purity of total RNA samples were checked by Nanodrop 1000 optical photometer. In each group, equal amount of purified total RNA from four dogs were pooled and then used for microarray analysis.
|
| Label |
Cy3
|
| Label protocol |
Agilent one-color Low Input Quick Amp Labeling Kit labeling protocol
|
| |
|
| Hybridization protocol |
Agilent one-color gene expression hyb/wash protocol
|
| Scan protocol |
Microarray slides were scanned in an Agilent DNA Sure Scan G4900DA at 3 micron resolution.
|
| Description |
Gene expression in small intestinal mucosa of 1st-infection group after 6days infection.
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 11.5.1.1 (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
Oct 17, 2017 |
| Last update date |
Jan 23, 2018 |
| Contact name |
Hirokazu Kouguchi |
| E-mail(s) |
kouguchi@d2.dion.ne.jp
|
| Organization name |
Hokkaido Institute of Public Health
|
| Street address |
N19, W12, Kita-ku
|
| City |
Sapporo |
| ZIP/Postal code |
0600819 |
| Country |
Japan |
| |
|
| Platform ID |
GPL15379 |
| Series (1) |
| GSE105098 |
Gene expression profile of small intestinal mucosa in dog repeatedly infected with the cestode Echinococcus multilocularis |
|
