|
| Status |
Public on Dec 14, 2007 |
| Title |
Lip-C-treated sham OBX C57 mouse, biol rep1 |
| Sample type |
RNA |
| |
|
| Source name |
microdissected olfactory epithelium
|
| Organism |
Mus musculus |
| Characteristics |
Genotype: w/t; strain C57BL/6J; male, 6 weeks old, olfactory epithelium
|
| Treatment protocol |
To deplete OE macrophages, mice were anesthetized with isoflurane and injected intranasally (i.n.) with 50 µl of liposomes containing clodronate (Lip-C) once per day at 4 PM for three consecutive days. To deplete a subpopulation of blood monocytes, the mice were injected intravenously (i.v.) with 200 µl Lip-C in the lateral tail vein on the first and third days of i.n. injections at the same time of day. Mice underwent bilateral sham olfactory bulbectomy on the second day of i.n. injections. Mice were euthanized at 48 h following sham OBX. Following euthanasia with carbon dioxide, the olfactory epithelium was quickly microdissected, weighed, and snap-frozen in liquid nitrogen.
|
| Growth protocol |
The mice were purchased from Jackson Laboratories (Bar Harbor, ME) at 5 weeks of age and were maintained in an animal facility at the University of Kentucky Department of Laboratory Animal Research (DLAR) until they reached 6 weeks of age. At the DLAR, they were kept on a 12 hr light:dark cycle in Bioclean units with sterile-filtered air and provided food and water ad libitum.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Snap-frozen microdissected olfactory epithelium was pulverized in TRI Reagent (Sigma-Aldrich, St. Louis, MO) and processed through a QIAshredder column (Qiagen, Valencia, CA). The total RNA was further purified using the Qiagen RNeasy Mini-Kit according to the manufacturer’s protocol. Total RNA yield and purity was assessed with a spectrophotometer and with the model 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA); all samples had A260/A280 ratios of 1.9–2.2 and showed two sharp peaks corresponding to 18S and 28S RNA on the Bioanalyzer electropherograms.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA was prepared according to the standard Affymetrix protocol from 3 ug total RNA (Expression Analysis Technical Manual, 2004, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 13.33 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Mouse Genome 430 2.0 Array . GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GCS 3000 7G scanner
|
| Description |
Gene expression data from olfactory epithelium of Lip-C-treated sham OBX C57 mice
|
| Data processing |
The data were analyzed with Affymetrix MicroArray Suite 5.0 using Affymetrix default analysis settings. A threshold value of 1,500 was used to normalize the data across all the microarrays.
|
| |
|
| Submission date |
Dec 15, 2006 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Marilyn L. Getchell |
| E-mail(s) |
mgetch@uky.edu
|
| Organization name |
University of Kentucky
|
| Department |
Anatomy and Neurobiology
|
| Lab |
Sanders-Brown Center on Aging
|
| Street address |
800 South Limestone St.
|
| City |
Lexington |
| State/province |
KY |
| ZIP/Postal code |
40536-0230 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE6540 |
Expression data from olfactory epithelium of Lip-C-treated mice compared to Lip-O-treated control mice |
|
| Relations |
| Reanalyzed by |
GSE119085 |
