Proteomic study of different epididymal segments in high-altitude Tibetan sheep
Ontology highlight
ABSTRACT:
ORGANISM(S): Ovis Aries
SUBMITTER: 
Xinrong Wang
PROVIDER: PXD072899 | iProX | Fri Jan 09 00:00:00 GMT 2026
REPOSITORIES: iProX
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Project description:While assisted reproductive technologies (ARTs) are widely used in domestic animals, successful implementation of ARTs to conserve wildlife species remains challenging. In macropods, crucial aspects of fundamental reproductive biology, including changes induced by epididymal maturation, remain unknown, limiting the development of ARTs. In this context we performed a proteomic analysis of spermatozoa from the caput, corpus, and cauda epididymis of Eastern Grey Kangaroos (n = 6) to profile changes over epididymal maturation. Samples prepared by FASP digestion were analysed by LC-MS/MS with SWATH acquisition. A total of 4,304 proteins were identified, with significant overlap across epididymal regions. Highly abundant proteins in common across caput, corpus and cauda spermatozoa had strong enrichment for tubulins and included 4 histone proteins. The most significant proteomic remodelling was observed in the corpus to cauda transition, late in epididymal transit (728 differentially abundant proteins). Overall proteomic changes across epididymal maturation (1,131 differentially abundant proteins) suggested a loss of sperm glycosidases and an increase in flagellar proteins, including tubulins and dyneins. These findings serve to highlight both consistencies with eutherian sperm epididymal maturation (e.g. bias towards protein loss over transit, transfer of proteins via extracellular vesicles) and elements which are likely unique to marsupials (e.g. reduced chromatin stability, potential use of β-oxidation as a major metabolic pathway). This critical information can now be leveraged to further develop ARTs in marsupials.
2025-09-29 | PXD064577 | Pride
Project description:<p>The epididymis is the primary organ responsible for sperm maturation, with its caudal region serving as the primary storage site for mature sperm. The unique small molecule metabolites present in the cauda epididymal fluid not only facilitate the long-term storage of sperm but also ensure their structural and functional integrity prior to fertilization. Consequently, an in-depth investigation into the composition and functions of these metabolites in the cauda epididymal fluid is essential for elucidating the regulatory mechanisms governing sperm maturation and fertilization. Yaks, as livestock unique to the Qinghai-Tibet Plateau, hold significant scientific and economic value. Compared to cattle, male yaks exhibit poorer reproductive performance, characterized by reduced sperm motility, elevated sperm abnormality rate, and lower artificial insemination conception rates. To investigate the relationship between sperm motility defects in yaks and the microenvironment during their maturation, this study collected cauda epididymal fluid from yaks and cattle for untargeted metabolomic sequencing. The results indicated that 1,098 and 1,297 kinds of metabolites annotated by the Human Metabolome Database (HMDB) database were identified in yak and cattle cauda epididymal fluid, respectively, using positive and negative ion modes. In both modes, the three categories containing the most types of metabolites were organic acids and derivatives, organoheterocyclic compounds, and lipids and lipid-like molecules. Within the lipid metabolites, fatty acids and conjugates were the most prevalent in both modes. In the positive ion mode, compared to cattle cauda epididymal fluid, 79 metabolites were significantly upregulated and 212 were significantly downregulated in yak cauda epididymal fluid. In the negative ion mode, compared to cattle cauda epididymal fluid, 110 metabolites were significantly upregulated and 230 metabolites were significantly downregulated in yak cauda epididymal fluid. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment results showed that the most significantly enriched pathway in the positive ion mode was the biosynthesis of amino acids, while in the negative ion mode, it was ABC transporters. Among the significant differentially expressed metabolites, 18 have been associated with sperm mobility, maturation or function. The functions of these metabolites involve regulation of oxidative stress, sperm capacitation, spontaneous acrosome reaction, mitochondrial energy metabolism, mRNA methylation, and flagellar motility. Taken together, these findings establish a foundational dataset and offer novel perspectives for elucidating the molecular mechanisms underlying yak sperm maturation, enhancing sperm motility, optimizing reproductive techniques, and ultimately improving reproductive performance.</p>
2025-09-29 | MTBLS13067 | MetaboLights
Project description:Following their production in the testis, spermatozoa enter the epididymis to gain their motility and fertilizing abilities. This post-testicular maturation coincides with sperm epigenetic profile changes that influence the progeny outcome. While recent studies underscored the dynamics of small non-coding RNAs in the maturing spermatozoa, little is known regarding sperm methylation changes and their impact at the post-fertilization level. To map out the sperm methylome dynamics, we purified spermatozoa by FACS from the testis and the different epididymal segments (i.e. caput, corpus and cauda) of CAG/su9-DsRed2; Acr3-EGFP transgenic mice. Reduced-Representation Bisulfite Sequencing (RRBS-Seq) performed on DNA from these respective sperm populations indicated that high methylation changes were observed between spermatozoa from the caput vs. testis with 5546 entries meeting our threshold values (q value < 0.01, methylation difference above 25 %). Most of these changes were transitory during epididymal sperm maturation according to the low number of entries identified between spermatozoa from cauda vs. testis.
2022-03-10 | GSE191051 | GEO
Project description:Sperm maturation depends on exposure to specific microenvironments within the different segments of the epididymis, but mechanisms underlying how these microenvironments are produced or maintained are not well understood. We hypothesize that epididymal extracellular vesicles (EVs) could play a role in this process, and therefore set out to test whether the EVs from different regions of the epididymis can serve as a form of paracrine communication between epithelial cells. Domestic cat tissues were used to develop a reproducible in vitro culture system for corpus epididymis explants that were then exposed to EVs collected from upstream (i.e., caput) segments. The impacts of different culture or exposure conditions were compared by analyzing the morphology, apoptosis, transcriptional activity, and gene expression in the explants. Here, we report the development of the first in vitro culture system for epididymal tissue explants in the domestic cat model. Using this system, we found that EVs from the caput segment have a significant effect on the transcriptional profile of tissue from the corpus segment (1,233 differentially expressed genes due to EV supplementation). Of note, expression of genes associated with regulation of epithelial cell differentiation and cytokine signaling in the epididymis were regulated by the presence of EVs. Together, our findings comprise the first report of paracrine control of segmental gene regulation by epididymal EVs in any species. These results contribute to a better understanding of epididymis biology and could lead to techniques to enhance or suppress male fertility.
2024-09-01 | GSE269443 | GEO
Project description:Tibetan's adaptation to high-altitude environment at the Qinghai-Tibetan plateau represents a remarkable case of natural selection during recent human evolution. We generated time series paired RNAseq, ATACseq and Hi-C data in Tibetan and Han Chinese's umbilical endothelial cells from normoxia to hypoxia condition. Our results provide a broad resource of genome-wide hypoxia regulatory network to characterize the effect of genetic variation in high-altitude adaptation, and indicates large-scale maps of variants need proper cell types to understand its act on gene regulation.
2020-07-15 | GSE145774 | GEO
Project description:Dicer1 is an endoribonuclease involved in the biogenesis of functional microRNAs (miRNAs). These small non-coding RNAs are important regulators of the posttranscriptional gene expression and participate to the control of male fertility. Knowing that 1) Dicer1-dependent factors are needed for proper sperm maturation in the epididymis, and that 2) miRNAs are potent mediators of intercellular communication in most biological systems, we investigated the role of Dicer1 dependent-miRNAs produced from the proximal epididymis (initial segment/caput) on the paracrine regulation of epididymal gene expression in the distal epididymis regions (i.e. corpus and cauda). To this aim, we performed comparative microarray and ANOVA analyses on control vs. Defb41iCre/wt;Dicer1fl/fl mice in which functional Dicer1 is absent from the principal cells of the proximal epididymis. We identified 21 and 16 transcripts, including Prostate And Testis Expressed 4 protein and the Zn-alpha 2-glycoprotein, that display significant expression level changes in the corpus and cauda regions, respectively (Fold change >2 or <-2; p<0.001). In addition, 154 miRNAs, including miR- 210, miR-672, miR-191 and miR-204, show a significantly impaired biogenesis in the absence of Dicer1 from the proximal epididymis (Fold change>2 or <-2; p<0.01). These miRNAs are secreted via extracellular vesicles derived from DC2 epididymal principal cell line, and their expression correlates with target transcripts involved in important biological pathways, as evidenced by in silico analysis. These observations suggest that Dicer1-dependent miRNAs could act as potent paracrine regulators of epididymal functions (i.e. control of sperm motility, ciliogenesis) and may contribute to the infertility phenotype observed in the Defb41iCre/wt;Dicer1fl/fl mouse model. These findings open new avenues for the identification of molecular targets important to male fertility control.
2016-09-19 | GSE77139 | GEO
Project description:This study investigates the changes in proteins and metabolites of sheep sperm across three regions of the epididymis: caput, corpus, and cauda.
2025-01-15 | PXD059850 |
Project description:Study question: Can region-specific transcriptional profiling of the epididymis from fertile and sub-fertile bulls predict the aetiology of fertility/subfertility in males? Summary answer:The highly regulated gene expression along the bovine epididymis is affected by the fertility status of bulls used for artificial insemination. What is known already: In mammals, sperm maturation and storage occur in the epididymis. Each epididymal segment has his own transcriptomic signature that modulates the intraluminal composition that governs sequential modifications of the maturing male gamete. Main results and the role of chance: Hierarchical clustering and Principal Component Analysis revealed a clear separation between caput, corpus and cauda epididymides. Some transcripts characterize a particular anatomical segment, whereas others are expressed in two out of three epididymal segments. GO analysis allowed deduction of specific functions played by each epididymal segment. The transcriptional profiles between fertile vs. sub-fertile conditions clustered most closely in the corpus and cauda segments, whereas the profiles in the caput segment were distinct between fertile and sub-fertile bulls. Of the differently expressed genes, ten (including AKAP4, SMCP, SPATA3, TCP11, ODF1, CTCFL, SPATA18, ADAM28, SORD and FAM161A) were found to exert functions related to reproductive systems and 5 genes (including DEAD, CYST11, DEFB119, DEFB124 and MX1) were found to be associated with the defence response. Limitations, reasons for caution: Further work is required to correlate these modulations of epididymal functions with sperm fertilizing ability in order to understand the aetiology of certain cases of idiopathic infertility in livestock and men. Wider implications of the findings: As fertility can be quantified in bulls used for artificial insemination, this species is a unique model to aid in the understanding of male fertility/subfertility in man.Our data provide a molecular characterization that will facilitate advances in understanding the involvement of epididymalphysiology in sub/infertility aetiology.
2017-03-31 | GSE96602 | GEO
Project description:In this study, we conducted a comprehensive quantitative phosphoproteomic analysis of mouse epididymal sperm from different regions (caput, corpus, and cauda) to unveil the dynamics of protein phosphorylation during sperm maturation. And we performed phosphoproteomic analysis of ALK inhibitor 2 compound 18-treated and control sperm to gain further insights into the potential mechanisms by which TSSK2 regulates sperm motility.
2024-10-17 | PXD049067 | Pride
Project description:Mammalian spermatozoa acquire their fertilizing ability during epididymal transit. Gene expression patterns along the epididymis are established by specific transcription factor networks that coordinate region-specific functions. The epididymis is usually divided into 3 segments: caput, corpus, and cauda. The human epididymis anatomy does not allow clear distinction between these three segments. To determine to which extent gene expression is segmented along the human epididymis, transcriptome profiling was performed on 8 distinct epididymal regions from 3 donors. Microarray analysis was performed on a Gene Chip Human Clariom S (Affymetrix®) array representing 337 100 transcriptional variants encoded by 20 800 genes. Proximal segments 1 to 3 were distinguishable from the distal epididymal segments (4 to 8) as shown by unsupervised Principal Component Analysis. Transcripts from each segment with differentially expressed genes (DEGs) > 2-fold change and FDR < 0.05 were clustered in relation to their intensity profiles. While no DEGs were detected between segments 1–3 corresponding to the efferent ducts, 1140 DEGs were detected between efferent ducts (1–3) and the epididymis (4–8), 400 between caput (4–6) vs. corpus/cauda (7–8) and none between corpus (7) and cauda (8). Gene Ontology annotation revealed that up-regulated DEGs in the efferent ducts (1–3) were predominantly related to cilium assembly/movement and cell differentiation. The biological process terms fertilization, defense and immune responses were associated with caput epididymis (4–6) while spermatogenesis and protein binding were found all along the epididymis (4–8). In conclusion, the proximal human epididymis is exclusively occupied by efferent ducts with a distinct DEG profile compared with the downstream epididymal segments. Moreover, gene expression profiling revealed two regions in the human epididymis; the caput and the distal corpus/cauda region. Taken together, analysis of the human epididymal transcriptome reveals a limited DEG profile.
2020-11-06 | GSE141568 | GEO