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 Affymetrix MG U74A v.2 microarrays.
Extracted molecule
total RNA
Label
Affymetrix labelling 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
Affymetrix data were filtered for spot quality by restricting analysis to genes with signal intensity greater than 100 for every sample.
