Identification of Differentially Expressed Gene Transcripts in Porcine Endometrium during Early Stages of Pregnancy.

Mariusz Pierzchała,Kamila Stepanow,Urszula Czarnik,Magdalena Herudzińska,Agnieszka Roszczyk,Dorota Pierzchała,Aleksandra Ciepłoch,Haja N Kadarmideen,Wojciech Niżański,Agata Nawrocka,Magdalena Ogłuszka,Tadeusz Blicharski,Magdalena Buszewska-Forajta,Mateusz Sachajko,Bartosz M Jaśkowski,Leyland Fraser,Ewa Poławska,J M Jaśkowski ,Brygida Ślaska ,Paweł Urbański ,A Korwin‐Kossakowska ,M.f.w Te Pas ,Chandra Shekhar Pareek

doi:10.3390/life10050068

Abstract

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 (p < 0.001; >2 FC). GO term enrichment analysis identified commonly shared upregulated endometrial DEG-transcript-probes (p < 0.001; >2 FC), that were regulating the gene functions of anatomic structure development and transport (TG), DNA-binding and methyltransferase activity (ZBTB2), ion-binding and kinase activity (CKM), cell proliferation and apoptosis activity (IL1B). Downregulated DEG-transcript-probes (p < 0.001; >2 FC) were involved in regulating the gene functions of phosphatase activity (PTPN11), TC616413 gene-transcript and Sus-scrofa LOC100525539. Moreover, blastn comparison of microarray-probes sequences against sus-scrofa11 assembly identified commonly shared upregulated endometrial DEG-transcript-probes (E < 0.06; >2 FC), that were regulating the gene functions of reproduction and growth (SELENOP), cytoskeleton organization and kinase activity (CDC42BPA), phosphatase activity (MINPP1), enzyme-binding and cell-population proliferation (VAV3), cancer-susceptibility candidate gene (CASC4), cytoskeletal protein-binding (COBLL1), ion-binding, enzyme regulator activity (ACAP2) Downregulated endometrial DEG-transcript-probes (E < 0.06; >2FC) were involved in regulating the gene functions of signal-transduction (TMEM33), catabolic and metabolic processes (KLHL15). Microarray validation experiment on selected candidate genes showed complementarity to significant endometrial DEG-transcript-probes responsible for the regulation of immune response (IL1B, S100A11), lipid metabolism (FABP3, PPARG), cell-adhesion (ITGAV), angiogenesis (IL1B), intercellular transmission (NMB), cell-adhesion (OPN) and response to stimuli (RBP4) 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.

Highlights

Reproduction is an essential element of effective animal production because it has a direct impact on its profitability
The quality analysis of endometrial microarray data were represented by the grouping of early proliferator‐activated gamma receptor), RBP4 (Retinol Binding Protein) and S100A11 (S100A11 protein) stages of pregnancies based on the expression signals across analyzed endometrial transcriptome were selected to validate the microarray experimental results (Table 3)
Results revealed the characteristics the grouping all three stages ofofearly probes were associated with regulation of significant of biologic processes,ofsuch as regulation pregnancies in Polish large-white (PLW) gilts based on their expression signals across analyzed endometrial transcriptome samples (Figure 3, Table S1)

Summary

Introduction

Reproduction is an essential element of effective animal production because it has a direct impact on its profitability. The crucial point is to create an optimal environment for the maturation of reproductive cells during the estrus cycle, followed by fertilization and fetal development. During the female estrous cycle, as well as during the early pregnancy periods in the reproductive system, there are physiological changes controlled at several levels, including gene expression changes in the endometrium [1,2]. The pregnancy in pigs takes about 114 days, while implantation, like in other livestock, such as a cow or sheep, is non-invasive [3,4]. In the first few weeks of pregnancy in the pig, these critical changes could be defined as the three main stages: (i) the peri-implantation period after fertilization, i.e., early pregnancy before maternal recognition of pregnancy (gestation period duration of 1–10 days: in this study 9th day), (ii) the peri-implantation period of maternal recognition of pregnancy (gestation period duration of 11–13 days: in this study 12th day) [6,7,8] and,

Results

Discussion

Conclusion