Analysis of the transcriptome of bovine endometrial cells isolated by laser micro-dissection (2): impacts of post-partum negative energy balance on stromal, glandular and luminal epithelial cells

Abstract

Background In post-partum dairy cows, the energy needs to satisfy high milk production induces a status of more or less pronounced Negative Energy Balance (NEB). NEB associated with fat mobilization impairs reproductive function. In a companion paper, we described constitutive gene expression in the three main endometrial cell types (stromal, glandular and luminal epithelial cells) isolated by laser capture micro-dissection (LCM) showing the specificities of their transcriptomic profiles. This study investigates the specific impact of NEB on gene expression in these cells around 80 days after parturition at day 15 of the oestrus cycle and describes their specific response to NEB. Results Following the description of their constitutive expression, the transcriptome profiles obtained by RNA sequencing of the three cells types revealed that differences related to the severity of NEB altered mainly specific patterns of expression related to individual cell types. Number of differentially expressed genes between severe NEB (SNEB) and mild NEB (MNEB) cows was higher in ST than in LE and GE, respectively. SNEB was associated with differential expression of genes coding for proteins involved in metabolic processes and embryo-maternal interactions in ST. Under-expression of genes encoding proteins with functions related to cell structure was found in GE whereas genes encoding proteins participating in pro-inflammatory pathways were over-expressed. Genes associated to adaptive immunity were under-expressed in LE. Conclusion The severity of NEB after calving is associated with changes in gene expression around 80 days after parturition corresponding to the time of breeding. Specific alterations in GEs are associated with activation of pro-inflammatory mechanisms. Concomitantly, changes in the expression of genes encoding proteins involved in cell interactions and maternal recognition of pregnancy takes place in ST. The combination of these effects possibly altering the uterine environment and embryo maternal interactions may negatively influence the establishment of pregnancy.

Progesterone differentially affects the transcriptomic profiles of cow endometrial cell types

“…This is particularly relevant for epithelial cells for which the cell type’s representability is low, as specific gene expression data is diluted in the average of the whole tissue. Furthermore, as observed here and recently also reported in another model [ 14 ], genes may only be differentially expressed in a specific cell type and remain unaffected in the others.…”

“…This study confirms the difficulty in capturing samples with good quality RNA from LE. As in Chankeaw et al [ 13 , 14 ], despite preparation of a high number of slides from biopsies, the number of sequenced samples was lower for LE than for GE and ST cells.…”

“…This method has been used to study the interactions between the endometrial epithelial and stromal compartments with seminal plasma in a murine model [ 9 ] and the conceptus induced regulation of endometrial function in the porcine [ 10 ], ovine [ 11 ] and equine [ 12 ] species. In the cow, recent studies based on this approach described the specific molecular signatures of endometrial stromal, glandular and luminal epithelial cells, as well as the effect of negative energy balance on the transcriptomic profiles of endometrial compartments in the mid-luteal phase [ 13 , 14 ]. The above experiments revealed that the different endometrial cell types show distinct transcription patterns.…”

Multi-Tissue Transcriptome Study of Innate Immune Gene Expression Profiling Reveals Negative Energy Balance Altered the Defense and Promoted System Inflammation of Dairy Cows

“…The RNA-seq next-generation sequencing (NGS) data of negative energy balance and ketosis-associated transcriptome in dairy cows was obtained from a public database (GEO database). The original sequencing data for liver, adipose, hypothalamus, endometrial, and blood corresponded to the following GEO numbers: 24 adipose samples [ 20 ] ( ; accessed on: 8 October 2020), 23 peripheral blood leukocyte samples [ 21 ] (PRJNA605719), 24 Endometrial samples [ 22 ] ( ; accessed on: 25 March 2020), 21 hypothalamus samples [ 23 ] ( ; accessed on: 5 August 2013), 18 liver samples [ 24 ] ( ; accessed on: 24 April 2012). The raw fasta data from all the samples were used for the subsequent analysis.…”

Subclinical endometritis differentially affects the transcriptomic profiles of endometrial glandular, luminal, and stromal cells of postpartum dairy cows

“…The bovine endometrium is a complex and heterogeneous tissue, and quantification of gene transcription from the whole endometrium may not reflect cell-specific transcription. In postpartum dairy cows, stromal (ST), glandular epithelial (GE), and luminal epithelial (LE) endometrial cells isolated by laser capture microdissection (LCM) exhibit very distinct molecular signatures (Chankeaw et al, 2021a), differentially influenced by progesterone (Pereira et al, 2022) and negative energy balance (Chankeaw et al, 2021b).…”