Series includes pooled (n = 5 mice per biological replicate) samples from lacrimal, meibomian, and submandibular glands of female palcebo- and testosterone-treated BALB/c mice. All mice were ovariectomized at 8 weeks and allowed to recover from surgery for 10 days prior to hormone implanatation. All experiments were run in triplicate (pooled biological replicates) on CodeLink Mouse Uniset I Microarrays.
Extracted molecule
total RNA
Label
CodeLink Expression Assay Reagent Kit
Description
Young adult female BALB/c mice (n = 30), ovariectomized at age 8 weeks, were purchased from Taconic Laboratories (Germantown, NY) and maintained in constant temperature rooms with fixed light/dark periods of 12 hours. Animals were allowed to recover from surgery for 10 days, then anesthetized with ketamine and xylazine, and subcutaneous implants of placebo (n = 15; cholesterol, methylcellulose, lactose)- or testosterone (n = 15; 10 mg)-containing pellets were inserted in the subscapular region. Hormone pellets were obtained from Innovative Research of America (Sarasota, FL) and were designed to maintain male physiologic amounts of testosterone over a two week period. Following two weeks of hormone exposure, the mice were sacrificed in a CO2 chamber, and the lacrimal, meibomian, and submandibular glands were removed and pooled (5 mice per group) into three groups by treatment (n = 6 groups). Glands were immediately frozen and stored in liquid nitrogen until RNA isolation.
Data processing
The 10,000 spot intensities on the scanned microarray image were normalized to a median value of 1 and data were exported for analysis.
