GSE136102
Profile of new early pregnancy markers identified by transcriptomic analysis in peripheral blood immune cells in beef heifers
We aimed with this study through RNAseq, found diferentially expressend genes (DEGs) that can be used as a new pregnancy markers in peripheral blood mononuclear cells (PBMC) on day 18 post artificial insemination; and to assess the mRNA profile by RT-qPCR of new pregnancy markers in PBMC and peripheral blood polymorphonuclear cells (PMN) at early pregnancy Methods: Nelore heifers (N=21) were artificially inseminated in a fixed time (D0). On Day 10, 14, 16, 18 and 20, blood was collected for isolation of PBMC and PMN, and P4 concentration assay and color Doppler ultrasonography was performed to evaluate the corpus luteum (CL) . Pregnancy diagnosis was done on Day 28 and heifers were classified in pregant (N=9) and non-pregant (N=12). Heifers (N=6/group) with different (P<0.05) plasma P4 concentration, CL area and blood perfusion on Day 18 were selected and RNAseq was done on PBMC samples. RNAseq analysis indicated 220 DEGs (200 up regulated in P). Twenty DEGs found on RNAseq of PBMC with the highest fold-changes or no overlapping between pregant  and non-pregant, were assessed by RT-qPCR from Day10 to 20 (N=6/group). Conclusions: nine DEGs studied by RT-qPCR presented increased expression on PBMC for Pregnant group in at least one-time point from Day16 to 20; but only 4 of these DEGs retained the expression increased on PMN (IFI6, IFI44, RSAD2 and LGALS3BP). Thus, results indicate potential genes to be used as novel pregnancy predictors in immune cells in cattle at early gestation. Overall design: The PBMC of six pregnant and six non-pregnant Nelore heifers on day 18 after artificial insemination were sequencing by RNAseq