Tributyltin chloride alters the structural, genomic, and epigenomic integrity of postejaculatory mammalian sperm
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ABSTRACT:
PROVIDER: PRJNA1268224 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR33758528_1.fastq.gz | Fastqsanger.gz | |||
| SRR33758528_2.fastq.gz | Fastqsanger.gz | |||
| SRR33758529_1.fastq.gz | Fastqsanger.gz | |||
| SRR33758529_2.fastq.gz | Fastqsanger.gz | |||
| SRR33758530_1.fastq.gz | Fastqsanger.gz |
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Project description:A global top priority for ameliorating male factor infertility includes identification of environmental factors and mechanisms that impact sperm function. Detection of endocrine disruptors (EDs) in seminal plasma and within the female reproductive tract has created an urgent need to understand how environmental stressors alter postejaculatory sperm function. Tributyltin chloride (TBT) is a model organotin, ED and epigenetic modifier that causes reproductive disorders in testicular sperm; however, consequences of TBT exposure on postejaculatory sperm prior to fertilization remain unknown. Our previous data indicate that TBT-exposed sperm affects initial cleavage and lowers embryo development potential. However, identifying reasons for idiopathic male factor subfertility is challenged by lack of routine clinical assessment for subtle phenotypes that compromise sperm function and therefore remain undetected with current diagnostic tools. Results The present study was aimed at identifying structural, genomic, and epigenomic consequences of TBT exposure to postejaculatory sperm in a temporal manner reflective of sperm transport prior to fertilization within the female reproductive tract. Bovine sperm were exposed to TBT (0 ,1, 10, 100nM) for 24 h under non-capacitating conditions at 25°C followed by quantification of sperm kinematics at 37°C, DNA integrity, and methylation status. No differences in sperm kinematics or capacitation status were detected after 24 h. However, acrosome integrity was compromised at both 0 and 24 h (P ≤ 0.05). Sperm DNA integrity was also negatively affected after 24 h. The methylation levels of select loci were quantified by PacBio single-molecule real-time (SMRT) long-read high-fidelity sequencing. Whole genome methyl-seq revealed 750 differentially methylated regions (DMRs) associated with exposure to TBT for 24 h. IPA and GO analyses identified embryo development, cell signaling, and transcriptional regulation as the most relevant bio-functions of TBT altered DMRs. Conclusions Postejaculatory mammalian sperm exposure to TBT negatively affects parameters important for sperm function while altering DNA integrity and the methylation profile of promotor regions. Consequences of sperm exposure to TBT include cellular and molecular mechanisms that are important for sperm function but remain undetected by routine clinical analyses. These findings provided new insight towards environmental impacts on postejaculatory sperm function.
2025-05-27 | GSE298065 | GEO
Project description:Human sperm RNA-seq was performed and sequences used for the generation of a RNA integrity index capable of assessing samples with high RNA fragmentation. A two-prong approach was used including both manual and computational analysis. This identified a series of sperm RNAs that are consistently intact, and acceptable for use in determining sample RNA quality. These RNAs function in roles related to spermatogenesis and/or fertilization.
2022-04-26 | GSE201081 | GEO
Project description:Purpose: To identify alterations induced by developmental exposure to tributyltin at different stages of life Methods: Dams were treated with tributyltin during gestation and lactation in drinking water and pups were euthanized at different ages Results: We identified a number of changes in transcriptomic activity in 5 month, and ten month old pups after tributyltin exposure. Male animals exposed to tributyltin also developed fatty liver 5 months of age and adenomas of the liver at ten months of age.
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Project description:The Transcript Integrity Index (TII) provides a standard measure of sperm RNA
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Project description:A bacterial isolate capable of utilizing tributyltin chloride (TBTCl) as sole carbon source was isolated from estuarine sediments of west coast of India and identified as Pseudomonas stutzeri based on biochemical tests and Fatty acid methyl ester (FAME) analysis. This isolate was designated as strain DN2. Although this bacterial isolate could resist up to 3 mM TBTCl level, it showed maximum growth at 2 mM TBTCl in mineral salt medium (MSM). Pseudomonas stutzeri DN2 exposed to 2 mM TBTCl revealed significant alteration in cell morphology as elongation and shrinkage in cell size along with roughness of cell surface. FTIR and NMR analysis of TBTCl degradation product extracted using chloroform and purified using column chromatography clearly revealed biotransformation of TBTCl into Dibutyltin dichloride (DBTCl₂) through debutylation process. Therefore, Pseudomonas stutzeri strain DN2 may be used as a potential bacterial strain for bioremediation of TBTCl contaminated aquatic environmental sites.
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Project description:The calcium-binding protein spermatid-associated 1 (Cabs1) is a novel spermatid-specific protein. However, its function remains largely unknown. In this study, we found that a long noncoding RNA (lncRNA) transcripted from the Cabs1 gene antisense, AntiCabs1, was also exclusively expressed in spermatids. Cabs1 and AntiCabs1 knockout mice were generated separately (using Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-Cas9 methods) to investigate their functions in spermatogenesis. The genetic loss of Cabs1 did not affect testicular and epididymal development; however, male mice exhibited significantly impaired sperm tail structure and subfertility. Ultrastructural analysis revealed defects in sperm flagellar differentiation leading to an abnormal annulus and disorganization of the midpiece-principal piece junction, which may explain the high proportion of sperm with a bent tail. Interestingly, the proportion of sperm with a bent tail increased during transit in the epididymis. Furthermore, Western blot and immunofluorescence analyses showed that a genetic loss of Cabs1 decreased Septin 4 and Krt1 and increased cyclin Y-like 1 (Ccnyl1) levels compared with the wild type, suggesting that Cabs1 deficiency disturbed the expression of cytoskeleton-related proteins. By contrast, AntiCabs1-/- mice were indistinguishable from the wild type regarding testicular and epididymal development, sperm morphology, concentration and motility, and male fertility. This study demonstrates that Cabs1 is an important component of the sperm annulus essential for proper sperm tail assembly and motility.
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Project description:RNA-Seq technique was applied to investigate the effects of two semen collection methods (Pelleted vs Liquefied) and two sperm purification methods (SCLB vs PS) to the integrity of isolated RNAs at different perspectives. The same set of semen samples were applied to investigate the qualitative and quantitative effect of semen collection methods and sperm cell purification methods on sperm transcript profiling.
2013-12-09 | E-GEOD-43586 | biostudies-arrayexpress
Project description:Interventions: Group 1: The project aims to:
Statistically compare the fragment length of cell-free DNA in the blood plasma of 80 patients with a confirmed diagnosis of colorectal carcinoma in stages I-IV with the cfDNA fragment lengths of 50 healthy subjects.
For this purpose, residual amounts of blood plasma samples are used, which are collected anyway as part of theroutine examinations.
Cell-free DNA was isolated from the blood plasma and the fragment lengths are measured using qPCR.
The project ends with the publication of the data.
Group 2: Statistically compare the fragment length of cell-free DNA in the blood plasma of 50 healthy subjects with 80 patients with a confirmed diagnosis of colorectal carcinoma in stages I-IV.
The plasma samples from healthy volunteers were purchased (Central Biohub GmbH).
Primary outcome(s): The aim of this study is: 1) to analyze total cfDNA concentration in CRC patients with different histopathological stages (UICC I-IV). 2) to address the question whether the integrity index of cfDNA increases in CRC patients as a result of elevated necrotic degradation processes. Therefore, we intended to quantify short and long fragments of cfDNA via qPCR independent from total cfDNA levels by using equal template concentrations for CRC patients and healthy individuals. This approach may help to understand whether previously described diagnostic markers are either increased, decreased or unaltered in blood plasma of CRC patients compared to healthy individuals.
Study Design: Allocation: ; Masking: ; Control: ; Assignment: ; Study design purpose: basic science
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Project description:The goal of this RNA-Seq study was to determine Acinetobacter baumannii's transcriptiional response to sub-MIC concentrations of benzalkonium chloride in Acinetobacter baumannii. This RNA-seq data was then utilized to aide in the determination of the sub-MIC mechanism of action for benzalkonium chloride.
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