Dynamics in the expression of epigenetic modifiers and histone modifications in perinatal rat germ cells during de novo DNA methylation†

Rwigemera, Arlette; Omri-Charai, Rhizlane El; Lecante, Laetitia L; Delbès, Géraldine

doi:10.1093/biolre/ioaa206

Cited by 6 publications

(6 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To test the impact of exposures specifically in the developing male germ cells, sorted GFP-positive cells were used. We have previously shown that this strategy allows germ cell sorting with high purity, viability and enrichment in germ cell-specific transcripts ( Rwigemera et al, 2021 ). Gene expression was studied using the Affymetrix Rat Gene 2.0 ST microarray revealing hundreds of DEGs per group under statistical cut-off of p -value<0.05 and a 1.5-fold difference.…”

Section: Discussionmentioning

confidence: 99%

“…Transgenic Sprague-Dawley rats expressing germ cell-specific GFP (GCS-EGFP) ( Cronkhite et al, 2005 ) were used as previously described ( Rwigemera et al, 2021 ). Rats were housed on a 12L:12D cycle and were fed with commercial food (Teklad global 18% protein, Envigo, Madison, WI) and tap water ad libitum .…”

Section: Methodsmentioning

confidence: 99%

“…Cell pellets of 50,000 to 100,000 cells were thawed on ice for 5–10 min prior to extracting total RNA using the Arcturus PicoPure RNA isolation kit (ThermoFisher Scientific, Saint-Laurent, Quebec, Canada, #KIT204) as previously described ( Rwigemera et al, 2021 ). Following RNA quantification with the NanoDrop One (Thermo Scientific, Wilmington, DE, United States), the RNA integrity number (RIN) was determined using a bioanalyzer.…”

Section: Methodsmentioning

confidence: 99%

“…Yet these cells express sex steroid receptors early in development ( Rouiller-Fabre et al, 2015 ) and are potential direct targets of EDCs. Moreover, the window of fetal sensitivity corresponds to important germ cell development and events ( Manku and Culty, 2015 ; Rwigemera et al, 2021 ) associated with major epigenetic and gene expression changes ( Law and Oatley, 2020 ; Rwigemera et al, 2021 ). It is therefore important to further investigate the effects of EDCs during these windows of sensitivity specifically on germ cells.…”

Section: Introductionmentioning

confidence: 99%

“…Here, we were inspired by the methods used in Naciff et al ( Naciff et al, 2005 ), to further test the immediate effects of in utero exposure to estrogen receptor agonists on germ cell gene expression. To that end, we took advantage of a transgenic rat expressing the green fluorescent protein (GFP) specifically in germ cells ( Cronkhite et al, 2005 ) allowing efficient purification of rat perinatal germ cells ( Rwigemera et al, 2017 ; Rwigemera et al, 2021 ). EE2 and GE were selected as xenoestrogens with different potency and affinity for the two ERs ( Kuiper et al, 1997 ).…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Impact of in Utero Rat Exposure to 17Alpha-Ethinylestradiol or Genistein on Testicular Development and Germ Cell Gene Expression

2022

Self Cite

View full text Add to dashboard Cite

Although the decline in male fertility is believed to partially result from environmental exposures to xenoestrogens during critical developmental windows, the underlying mechanisms are still poorly understood. Experimental in utero exposures in rodents have demonstrated the negative impact of xenoestrogens on reproductive development, long-term adult reproductive function and offspring health. In addition, transcriptomic studies have demonstrated immediate effects on gene expression in fetal reproductive tissues, However, the immediate molecular effects on the developing germ cells have been poorly investigated. Here, we took advantage of a transgenic rat expressing the green fluorescent protein specifically in germ cells allowing purification of perinatal GFP-positive germ cells. Timed-pregnant rats were exposed to ethinylestradiol (EE2, 2 μg/kg/d), genistein (GE, 10 mg/kg/d) or vehicle by gavage, from gestational days (GD) 13–19; testes were sampled at GD20 or post-natal (PND) 5 for histological analysis and sorting of GFP-positive cells. While EE2-exposed females gained less weight during treatment compared to controls, neither treatment affected the number of pups per litter, sex ratio, anogenital distance, or body and gonadal weights of the offspring. Although GE significantly decreased circulating testosterone at GD20, no change was observed in either testicular histology or germ cell and sertoli cell densities. Gene expression was assessed in GFP-positive cells using Affymetrix Rat Gene 2.0 ST microarrays. Analysis of differentially expressed genes (DEGs) (p < 0.05; fold change 1.5) identified expression changes of 149 and 128 transcripts by EE2 and GE respectively at GD20, and 287 and 207 transcripts at PND5, revealing an increased effect after the end of treatment. Only about 1% of DEGs were common to both stages for each treatment. Functional analysis of coding DEG revealed an overrepresentation of olfactory transduction in all groups. In parallel, many non-coding RNAs were affected by both treatments, the most represented being small nucleolar and small nuclear RNAs. Our data suggest that despite no immediate toxic effects, fetal exposure to xenoestrogens can induce subtle immediate changes in germ cell gene expression. Moreover, the increased number of DEGs between GD20 and PND5 suggests an effect of early exposures with latent impact on later germ cell differentiation.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%