organism: Mus musculus
description: Protocols: We used 10 virgin female mice (Mus musculus L., outbred MF1) kept on a 12 h:12 h light:dark cycle (lights on 07:00·h) at ambient temperature of 21°C (range 20–22°C) and relative humidity of 59% (range 54–64%). Food (CRM, Pelleted Rat and Mouse Breeder and Grower Diet, Special Diets Services, BP Nutrition, Witham, Essex, UK) and water were available ad libitum. At age of 9–11 weeks, mice were acclimated to the single housing environment for 1 week, after which they were paired with males for 11 days. All females got pregnant and gave birth to young. The day of birth was counted as day 0 of lactation. Body mass, food intake, litter size and litter mass were recorded from day 4 to end of lactation (day 18). On day 6 of lactation, half of the lactating females (n = 5) were shaved, while the other half (n = 5) served as an unshaved control group (details below). Milk production was evaluated at peak lactation from measurements of metabolizable energy intake (MEI) and daily energy expenditure (DEE) (details below). On day 18 of lactation, mice were sacrificed by CO2 overdose, followed by the excision of the right inguinal mammary gland. The tissue was immediately frozen in liquid N2 and stored at −80°C prior to RNA extraction. Once the measurements on day 6 of lactation were completed, all 10 lactating females were anaesthetized with gaseous isoflurane for ~10 min. While under anaesthesia, 5 females were shaved dorsally (using a Wella Contura Hair Clipper, Basingstoke, Hants, UK) to gently remove ~72% of fur (Król et al. 2007). The remaining mice were handled similarly but not shaved. Hair regrowth was prevented by repeating the shaving protocol on days 10 and 14 of lactation. Total RNA from the inguinal mammary gland was isolated by homogenization of ~100 mg of tissue in TRIzol® Reagent (Ambion by Life Technologies, Carlsbad, CA, USA), using 3 mm tungsten carbide beads and a TissueLyser II Disruption System (Qiagen GmbH, Hilden, Germany). Following isolation, the RNA was quantified by spectrophotometry (NanoDrop Technologies, Wilmington, DE, USA) and its integrity was confirmed by electrophoresis (Agilent Technologies, Santa Clara, CA, USA). All RNA samples had a RIN number > 7.8, meeting the criteria for RNA-seq. RNA-seq library preparation and sequencing were carried at the Beijing Genomics Institute (BGI, Shenzhen, China). The libraries for each of the 10 samples were constructed using the TruSeq Stranded mRNA Sample Preparation Kit (Illumina, San Diego, CA), according to the manufacturer's instructions.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2022-12-31
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2022-12-31
attributes.tag: External Id
attributes.value: SAMEA14201046
attributes.tag: INSDC center alias
attributes.value: UNIVERSITYOFABERDEEN
attributes.tag: INSDC center name
attributes.value: University of Aberdeen
attributes.tag: INSDC first public
attributes.value: 2022-12-31T00:58:16Z
attributes.tag: INSDC last update
attributes.value: 2022-12-31T00:58:16Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-11654:Sample 6
attributes.tag: age
attributes.value: 9-11
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: individual
attributes.value: 6
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: milk production
attributes.value: 130.9 kilojoule per day
attributes.tag: organism part
attributes.value: mammary gland
attributes.tag: sample name
attributes.value: E-MTAB-11654:Sample 6
attributes.tag: sex
attributes.value: female
attributes.tag: stimulus
attributes.value: unshaved fur
attributes.tag: strain
attributes.value: MF1