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- Chankeaw, Wiruntita, et al.
(author)
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Analysis of the transcriptome of bovine endometrial cells isolated by laser micro-dissection (1): specific signatures of stromal, glandular and luminal epithelial cells
- 2021
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In: BMC Genomics. - : Springer Science and Business Media LLC. - 1471-2164. ; 22
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Journal article (peer-reviewed)abstract
- Background A number of studies have examined mRNA expression profiles of bovine endometrium at estrus and around the peri-implantation period of pregnancy. However, to date, these studies have been performed on the whole endometrium which is a complex tissue. Consequently, the knowledge of cell-specific gene expression, when analysis performed with whole endometrium, is still weak and obviously limits the relevance of the results of gene expression studies. Thus, the aim of this study was to characterize specific transcriptome of the three main cell-types of the bovine endometrium at day-15 of the estrus cycle. Results In the RNA-Seq analysis, the number of expressed genes detected over 10 transcripts per million was 6622, 7814 and 8242 for LE, GE and ST respectively. ST expressed exclusively 1236 genes while only 551 transcripts were specific to the GE and 330 specific to LE. For ST, over-represented biological processes included many regulation processes and response to stimulus, cell communication and cell adhesion, extracellular matrix organization as well as developmental process. For GE, cilium organization, cilium movement, protein localization to cilium and microtubule-based process were the only four main biological processes enriched. For LE, over-represented biological processes were enzyme linked receptor protein signaling pathway, cell-substrate adhesion and circulatory system process. Conclusion The data show that each endometrial cell-type has a distinct molecular signature and provide a significantly improved overview on the biological process supported by specific cell-types. The most interesting result is that stromal cells express more genes than the two epithelial types and are associated with a greater number of pathways and ontology terms.
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| 2. |
- Chankeaw, Wiruntita, et al.
(author)
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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
- 2021
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In: BMC Genomics. - : Springer Science and Business Media LLC. - 1471-2164. ; 22
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Journal article (peer-reviewed)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.
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| 4. |
- Chankeaw, Wiruntita, et al.
(author)
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Elevated non-esterified fatty acids impair survival and promote lipid accumulation and pro-inflammatory cytokine production in bovine endometrial epithelial cells
- 2018
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In: Reproduction, Fertility and Development. - 1031-3613 .- 1448-5990. ; 30, s. 1770-1784
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Journal article (peer-reviewed)abstract
- Elevated non-esterified fatty acids (NEFAs) are associated with negative effects on bovine theca, granulosa and oviductal cells but the effects of NEFAs on bovine endometrial epithelial cells (bEECs) are not as well documented. The objective of this study was to define the effects of NEFAs on bEECs. Postprimary bEECs were treated with 150, 300 or 500 mu M of either palmitic acid (PA), stearic acid (SA) or oleic acid (OA) or a mixture of NEFAs (150 mu M of each FA) or 0.5% final concentration of vehicle ethanol (control). Viability and proliferation of bEECs exposed to 150 mu M of each NEFA or a mixture of NEFAs were unaffected. Increased lipid accumulation was found in all treated groups (P < 0.01). In cells exposed to 500 mu M of each NEFA and 300 mu M PA decreased cell viability (P < 0.001), proliferation (P < 0.05) and increased apoptosis (P < 0.05) were observed. Treatment with 500 mu M OA, PA and SA had the strongest effects on cell viability, proliferation and apoptosis (P < 0.05). Treatment with PA and OA increased interleukin-6 (IL-6) concentrations (P < 0.05), whereas only the highest concentration of PA, OA and SA stimulated IL-8 production (P < 0.05). These results suggest that high concentrations of NEFAs may impair endometrial function with more or less pronounced effects depending on the type of NEFA and time of exposure.
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| 5. |
- Chankeaw, Wiruntita
(author)
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Impacts of fat mobilization on bovine endometrium function
- 2019
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Doctoral thesis (other academic/artistic)abstract
- In cows with high milk production, negative relationships between negative energy balance (NEB), inducing non-esterified fatty acids (NEFAs) in the peripheral circulation, and reproductive performances are well documented. Three major NEFAs; oleic (OA), palmitic (PA) and stearic acids (SA) have adverse effects on reproductive cells. In this thesis, the effects of NEB on gene expression in specific endometrial cells were studied from an in vivo model. Full endometrial biopsies were collected from Swedish Red Breed cows with different NEB status at 80 days postpartum. The three types of endometrial cells (stroma; ST, glandular epithelium; GE, and luminal epithelium; LE) were isolated from a tissue sample by using laser microdissection. Cell type-specific gene expression was identified in those cell types by RNA sequencing. The effects of the severity of NEB on endometrial gene expression were identified and it was found that the response to NEB was specific in the different cell types. Changes in gene expression are more profound in ST in which genes encoding proteins involved in energy metabolism, cell structure and peri-implantation processes were affected. Over-expression of genes associated with inflammatory response was observed in GE, whereas under-expression of genes related to adaptive immunity was observed in LE. In an in vitro study, we investigated the effects of the three NEFAs on phenotypic and gene expression responses of bovine endometrial epithelial cells (bEECs) in culture. Cultured postprimary bEECs were exposed to different concentrations of OA, PA and SA or their mixture. Cell viability, proliferation, apoptosis, lipid accumulation, pro-inflammatory cytokines (IL-6 and IL-8) production were examined at 24 and 48 h. Elevated NEFA dosage decreased cell viability and proliferation, while apoptosis, lipid accumulation and cytokine production were increased. The negative effects of OA and PA were stronger than those induced by SA. Gene expression changes of bEECs induced by 300 µM of either OA or PA for 2 h were further determined by using RNA sequencing. Oleic acid induced stronger changes in gene expression than PA. The up-regulation of pathways related to energy metabolism, apoptosis and oxidative stress signaling were observed whereas significance was not reached following use of PA. In conclusion, these results indicate that metabolic stress can affect endometrial functions in various ways. The gene expression changes in vivo suggests that severe NEB have long term effects on endometrial cells and that the responses are specific to cell types. The results from in vitro studies clearly show that NEFAs, especially OA and PA, alters many functions of bEECs and are detrimental to the sub-population of epithelial cells. Taken together, the results document that metabolic imbalance during the post-partum period alters several functions in the endometrium in a specific way in the different cell types and strongly stimulate pro-inflammatory mechanisms especially in epithelial cells.
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