Dataset Information

Endometrial gene expression in fertile and subfertile cows

ABSTRACT: In order to clarify the cause of reproductive failure, we conducted global endometrial gene expression analysis in fertile and subfertile cows. Hierarchical cluster analysis with the expression levels of mitochondrial DNA encoded genes was divided these cows into two clusters. One cluster was composed of fertile cows, the other cluster contained subfertile cows, and the expressions of mitochondrial DNA encoded genes in subfertile cows were higher than those in fertile cows

ORGANISM(S): Bos taurus

PROVIDER: GSE107741 | GEO | 2018/01/11

SECONDARY ACCESSION(S): PRJNA421264

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:The miRNA profiles were measured using small-RNA sequencing in beef heifers sampled at weaning that was retrospectively classified as fertile or subfertile following the breeding protocol. To accomplish this, the miRNA profiles were generated from the blood samples (10 mL) collected from crossbred heifers (Angus-Simmental) at the time of weaning (~238 days after birth). Peripheral white blood cells (PWBC) were extracted from the blood samples and stored at -80°C until further processing. During the breeding season, all the heifers followed the same breeding protocol, estrus synchronization, and fixed-time artificial insemination (AI). Fourteen days following the fixed-time AI, the non-pregnant heifers were exposed to fertile bulls for 60 days. Depending on the presence or absence of conceptus at 75 days following AI, heifers were classified as fertile for those who were pregnant through artificial insemination, pregnant to natural breeding (P-NB), or subfertile for those who were not pregnant. Heifers from fertile (n = 7) and subfertile (n = 7) groups were considered for the study. Total RNA was extracted from the PWBC of 14 samples and was subjected to small RNA library preparation and sequencing. After quality control, adapter trimming, and alignment, mature miRNAs were used for differential expression analysis. The read counts were transformed to counts per million (CPM), and raw counts with CPM < 1 in 50% of the samples were filtered out. The filtered raw counts were analyzed using DESeq2 v 1.26.0 to identify differentially expressed miRNAs (DEMIs). The DEMIs identified with a p-value < 0.05 and absolute (log2 fold change) > 0.5 were considered significant. With the subfertile heifers as the reference group, we identified 16 DEMIs between fertile and subfertile groups. To determine the genes targeting the DEMIs, we downloaded the target genes for each DEMI and retained only those genes expressed in the PWBCs. For the miRNA-gene correlation, we used the partial correlation and information theory (PCIT) approach to identify the significant gene-miRNA correlated pairs. The significant genes correlated with the miRNAs identified pathways including MAPK, ErbB, HIF-1, FoxO, p53, mTOR, T-cell receptor, insulin and GnRH signaling, apoptosis, and pathways regulating pluripotency of stem cells in the fertile group while cell cycle, p53 signaling pathway and apoptosis pathways in the subfertile group.

Project description:Thoroughbred stallions that carry a double-homozygous genotype A/A-A/A for SNPs rs397316122 and rs69101140 in exon 5 of the FKBP6 gene (chr13; EquCab3.0) are uniquely subfertile due to impaired acrosomal exocytosis (IAE). However, clear causation of this condition has not been determined. In the current study, the sperm proteome in frozen/thawed semen from three fertile TB stallions and three subfertile TB stallions (FKBP6 genotype A/A-A/A) was studied using mass spectrometry-based global proteomics. Sperm from both stallion groups were incubated in Lactate-Modified Whitten's (Lac-MW) medium to induce spontaneous acrosomal exocytosis in viable sperm (AE/Viable). At 0h, 2h, 4h, and 6h, sperm aliquots were removed for analysis for AE/Viable using flow cytometry (Fixable Live/Dead Red + FITC-PSA) global proteomics via data-independent acquisition mass spectrometry (DIA-MS). Student's t-tests and two-way ANOVA with Benjamini-Hochberg multiple testing correction (FDR q-value 0.05) were used to determine differences in AE/Viable and protein relative abundance between experimental groups and incubation periods. At 4h and 6h of incubation, the mean AE/Viable was higher in the fertile than in the subfertile stallions (41 and 44% vs. 14 and 16%, respectively; p < 0.05). A total of 2220 proteins was identified by DIA-MS. Using strict selection criteria (FDR 1.0%, q-value < 0.05, and fold-change < 1.5 or > 1.5), 140 proteins were found to be differentially abundant in sperm from the subfertile stallions when compared to that of the fertile stallions (83 less and 57 more abundant) at 0h incubation. Using bioinformatic analyses, most of the proteins of lower abundance in sperm from subfertile stallions were found to be overrepresented in the "metabolism" (32 proteins) and "metabolism of lipids" (18 proteins) pathways (Homo sapiens orthologs; Reactome database). Two of these proteins, arylsulfatase F (ARSF; log2 fold change = -2.0; p < 0.05) and zona pellucida-binding protein 2 (ZPBP2; log2 fold change = -1.7; p < 0.05), are acrosomal proteins known to play a fundamental role in sperm-oocyte binding. By using immunofluorescence, at 0h of incubation in MW-Lac, ARSF was identified at the acrosome, mid-, and principal piece in sperm from fertile TB stallions, while only at the mid-and principal piece in sperm from subfertile TB stallions. No evidence of ZPBP2 was observed in sperm from both stallion groups at 0h incubation in Lac-MW. Sperm from subfertile Thoroughbred stallions were bound to porcine zonae pellucidae at a lower frequency than sperm from fertile Thoroughbred stallions. In conclusion, DIA-MS is a powerful tool to identify candidate proteins that contribute to the etiology of IAE in Thoroughbred stallions, including proteins of acrosome origin that have a role in the sperm-oocyte binding process.

Dataset Information

Endometrial gene expression in fertile and subfertile cows

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets