A gene network of uterine luminal epithelium organizes mouse blastocyst implantation

RNA-seq analyses using embryos and uterine luminal layers collected from female mice on day 3 am, day 4 am, day 4 pm and day 5 am of pregnancy. Wildtype C57BL/6N females were used. The day when the vaginal plug was detected was considered day 1 of pregnancy. Pregnant mice were euthanized by cervical dislocation on designated days of pregnancy for the evaluation of pregnancy phenotypes and sample collection. On day 3, oviducts were flushed to confirm presence of 8 cells. On day 4, one uterine horn was flushed with saline to confirm the presence of blastocysts and the other side of the uterine horn was snap-frozen for cryosectioning. On day 5, mice were treated with 1% Chicago blue in saline i.v. to visualize implantation sites. Laser microdissection was performed to collect luminal layers.本研究针对妊娠第3日上午、第4日上午、第4日下午及第5日上午的雌性小鼠采集的胚胎与子宫腔上皮层开展RNA测序（RNA-seq）分析。实验采用野生型C57BL/6N品系雌性小鼠，以检测到阴道栓的当日记为妊娠第1日。在指定妊娠日期通过颈椎脱臼法处死孕鼠，以评估妊娠表型并采集样本。妊娠第3日时，冲洗输卵管以确认存在8细胞期胚胎；妊娠第4日，用生理盐水冲洗一侧子宫角以确认囊胚存在，另一侧子宫角则快速冷冻用于冷冻切片；妊娠第5日，经静脉注射（i.v.）1%生理盐水配制的芝加哥蓝溶液以显示着床位点，随后通过激光显微切割技术采集子宫腔上皮层。