GSM2068611: Pineal Dusk rep1; Macaca mulatta; RNA-Seq
Twelve adult rhesus macaques (Macaca mulatta, 3 to 33 years of age, male and female, 4 to 10.3 kg) were obtained through the Veterinary Resources Program Primate Recycle Program of NIH over a period of 11 years.     All protocols and handling of the animals conformed to the Guidelines for Care and Use of Laboratory Animals (NIH Publication 80-23; Protocols 99-012 and 11-055 ). Animals were housed under a 12-h light, 12-h dark lighting regimen (lights on at 0600 h) in NIH facilities for at least 6 wk. The light level in the cages was approximately 325 Lux.   Animals (three per time point) were sacrificed between 1100 and 1300 (day) , 1700 and 1900 (dusk), 2300 and 0100 (night) and 0500 and 0700 (dawn).  Animals were anesthetized (ketamine, 0.1 ml/kg body weight) and a catheter was placed in the saphenous vein prior to transport to the procedure room.   Animals were sacrificed using Euthasol (Virbac, Fort Worth, TX)  or Beuthanasia D ( Schering Plough, Union, NJ).   Animals sampled at night  or dusk  were anesthesized  under dim red light and  blindfolded; euthanasia was done under dim white light and  the blindfold was removed  after decapitation. Removal of tissues, including the pineal gland, retina, and others (corpus callosum, frontal cortex, cerebellum, thalamus, hippocampus, caudate, medulla, pituitary, heart, liver, lung, kidney, skeletal muscle, small intestine,  testis and  ovary) was complete within 45 minutes following euthanasia; immediately following removal, samples  of each tissue were obtained and placed on solid  CO2.  Tissue was store at -80° C. Total RNA was extracted from individual pineal glands or pieces of other tissues (  2 to 4 mm 3 pieces) with TRIzol reagent. RNA libraries were prepared for sequencing using standard Illumina protocols