Pierzchała Mariusz, Pierzchała Dorota, Ogłuszka Magdalena, Poławska Ewa, Blicharski Tadeusz, Roszczyk Agnieszka, Nawrocka Agata, Urbański Pawel, Stepanow Kamila, Ciepłoch Aleksandra, Korwin-Kossakowska Agnieszka, Pas Marinus F W Te, Slaska Brygida, Buszewska-Forajta Magdalena, Jaśkowski Jedrzej M, Sachajko Mateusz, Herudzińska Magdalena, Jaśkowski Bartosz M, Niżański Wojciech, Fraser Leyland, Czarnik Urszula, Kadarmideen Haja N, Pareek Chandra S
Department of Animal Genomics and Biodiversity, Institute of Genetics and Animal Breeding, Polish Academy of Sciences, Jastrzebiec, 05-552 Magdalenka, Poland.
Wageningen UR Livestock Research, Animal Breeding and Genomics, 6708 PB Wageningen, The Netherlands.
Life (Basel). 2020 May 16;10(5):68. doi: 10.3390/life10050068.
During the early stages of pregnancy, the uterine endometrium undergoes dramatic morphologic and functional changes accompanied with dynamic variation in gene expression. Pregnancy-stage specific differentially expressed gene (DEG)-transcript-probes were investigated and identified by comparing endometrium transcriptome at 9th day (9D), 12th day (12D) and 16th day (16D) of early pregnancy in Polish large-white (PLW) gilts. Endometrium comparisons between 9D-vs-12D, 9D-vs-16D and 12D-vs-16D of early pregnancy identified 6049, 374 and 6034 highly significant DEG-transcript-probes ( < 0.001; >2 FC). GO term enrichment analysis identified commonly shared upregulated endometrial DEG-transcript-probes ( < 0.001; >2 FC), that were regulating the gene functions of anatomic structure development and transport (), DNA-binding and activity (), ion-binding and activity (), cell proliferation and apoptosis activity (. Downregulated DEG-transcript-probes ( < 0.001; >2 FC) were involved in regulating the gene functions of activity ( TC616413 gene-transcript and LOC100525539. Moreover, blastn comparison of microarray-probes sequences against assembly identified commonly shared upregulated endometrial DEG-transcript-probes (E < 0.06; >2 FC), that were regulating the gene functions of reproduction and growth (), cytoskeleton organization and activity (), activity (), enzyme-binding and cell-population proliferation (VAV3), cancer-susceptibility candidate gene (), cytoskeletal protein-binding (), ion-binding, enzyme regulator activity () Downregulated endometrial DEG-transcript-probes (E < 0.06; >2FC) were involved in regulating the gene functions of signal-transduction ( catabolic and metabolic processes (. Microarray validation experiment on selected candidate genes showed complementarity to significant endometrial DEG-transcript-probes responsible for the regulation of immune response (), lipid metabolism (), cell-adhesion (), (), intercellular transmission (), cell-adhesion () and response to stimuli () was confirmed by RT-PCR. This study provides a clue that identified pregnancy-stage specific microarray transcript probes could be considered as candidate genes for recognition and establishment of early pregnancy in the pig.
在妊娠早期，子宫内膜会发生显著的形态和功能变化，并伴随着基因表达的动态变化。通过比较波兰大白（PLW）后备母猪妊娠早期第9天（9D）、第12天（12D）和第16天（16D）的子宫内膜转录组，研究并鉴定了妊娠阶段特异性差异表达基因（DEG）-转录本-探针。妊娠早期9D与12D、9D与16D以及12D与16D之间的子宫内膜比较，鉴定出6049、374和6034个高度显著的DEG-转录本-探针（<0.001；>2倍变化）。基因本体（GO）术语富集分析确定了共同上调的子宫内膜DEG-转录本-探针（<0.001；>2倍变化），这些探针调节解剖结构发育和运输、DNA结合和活性、离子结合和活性、细胞增殖和凋亡活性等基因功能。下调的DEG-转录本-探针（<0.001；>2倍变化）参与调节活性（TC616413基因-转录本和LOC100525539）等基因功能。此外，将微阵列探针序列与基因组组装进行blastn比较，确定了共同上调的子宫内膜DEG-转录本-探针（E<0.06；>2倍变化），这些探针调节生殖和生长、细胞骨架组织和活性、活性、酶结合和细胞群体增殖（VAV3）、癌症易感性候选基因、细胞骨架蛋白结合、离子结合、酶调节活性等基因功能。下调的子宫内膜DEG-转录本-探针（E<0.06；>2倍变化）参与调节信号转导、分解代谢和代谢过程等基因功能。对选定候选基因的微阵列验证实验表明，与负责调节免疫反应、脂质代谢、细胞粘附、细胞间传递、细胞粘附和对刺激的反应等重要子宫内膜DEG-转录本-探针具有互补性，并通过逆转录-聚合酶链反应（RT-PCR）得到证实。本研究提供了一条线索，即所鉴定的妊娠阶段特异性微阵列转录探针可被视为识别和确定猪早期妊娠的候选基因。
