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Testicular lumicrine factors regulate ERK, STAT and NFKB pathways in the initial segment of the rat epididymis to prevent apoptosis

ABSTRACT: The initial segment of the epididymis is vital for male fertility, therefore, it is important to understand the mechanisms that regulate this important region. Deprival of testicular luminal fluid factors/lumicrine factors from epididymis, a subset of cells within the initial segment undergo apoptosis. In this study, microarray analyses was used to examine early changes in the downstream signal transduction pathways following the loss of lumicrine factors, and we discovered the following cascade of events leading to loss of protection and eventual apoptosis. First, mRNA expression of several key components of ERK pathway decreased sharply after 6 hours of loss protection from testicular lumicrine factors. After 12 hours, the levels of mRNA expression of STAT and NF-кB pathways components increased, mRNA expression of genes encoding cell cycle inhibitors increased. After 18 hours of loss protection from testicular lumicrine factors, apoptosis was observed in the initial segment. In conclusion, testicular lumicrine factors protect the cells of the initial segment by activating ERK pathway, repressing STAT and NF-кB pathways, and preventing a cascade of reactions leading to apoptosis.

ORGANISM(S): Rattus norvegicus

PROVIDER: GSE21806 | GEO | 2011/03/11

SECONDARY ACCESSION(S): PRJNA126965

REPOSITORIES: GEO

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Testicular lumicrine factors regulate ERK, STAT and NFKB pathways in the initial segment of the rat epididymis to prevent apoptosis

Project description:The initial segment of the epididymis is vital for male fertility, therefore, it is important to understand the mechanisms that regulate this important region. Deprival of testicular luminal fluid factors/lumicrine factors from epididymis, a subset of cells within the initial segment undergo apoptosis. In this study, microarray analyses was used to examine early changes in the downstream signal transduction pathways following the loss of lumicrine factors, and we discovered the following cascade of events leading to loss of protection and eventual apoptosis. First, mRNA expression of several key components of ERK pathway decreased sharply after 6 hours of loss protection from testicular lumicrine factors. After 12 hours, the levels of mRNA expression of STAT and NF-кB pathways components increased, mRNA expression of genes encoding cell cycle inhibitors increased. After 18 hours of loss protection from testicular lumicrine factors, apoptosis was observed in the initial segment. In conclusion, testicular lumicrine factors protect the cells of the initial segment by activating ERK pathway, repressing STAT and NF-кB pathways, and preventing a cascade of reactions leading to apoptosis. Unilateral efferent duct ligation (EDL) surgeries were performed to block lumicrine testicular fluid factors from reaching one side of epididymis. For the control, a sham operation was performed on the contra-lateral side within the same animal. After 6, 12 and 18 hours of EDL, the most proximal region of the initial segment containing zone 1a and 1b of initial segment was dissected out. In this microarray study, sham control and each time point of EDL (sham, 6h EDL, 12h EDL, 18h EDL) all have 4 replicates, a total of 16 arrays were hybridized.

2011-03-11 | E-GEOD-21806 | biostudies-arrayexpress

Rattus norvegicus

Project description:Testicular lumicrine factors regulate ERK, STAT and NFKB pathways in the initial segment of the rat epididymis to prevent apoptosis

| PRJNA126965 | ENA

PTEN is essential in the maintenance of normal transporter activity in the epididymal epithelium

Project description:A fully developed initial segment, the most proximal region of the epididymis, is important for male fertility. Conditional deletion of Pten from the initial segment from postnatal day 17 onwards resulted in de-differentiation of the initial segment. When spermatozoa progressed through the de-differentiated epididymal duct, they developed angled flagella, suggesting compromised sperm maturation, which eventually resulted in male infertility. To understand mechanisms that underlie this sperm maturation defect following loss of Pten, we compared transcriptome profile of the initial segment between controls and knockouts and revealed that the transporter activity was one of the top molecular and cellular functions altered following loss of Pten. Alteration in the protein levels and localization of several transporters following loss of Pten were also observed by immunofluorescence analysis. Epithelial cells of the initial segment from knockouts are more permeable to Fluorescein isothiocyanate-dextran (4000da) compared to controls. When analyzing GEO datasets with Pten deletion in the prostate and inner ear of mice, alteration in transporter activity was also found in these Pten knockout mouse models, suggesting it is a common phenomenon. PTEN regulates transporter activities probably through regulation of cell shape, size, and membrane property, which in turn regulates the intracellular and intercellular micro-environment.

2019-12-19 | GSE92614 | GEO

Anatomical transcriptome atlas of the male mouse reproductive system during aging

Project description:The testis, efferent ductules, epididymis, and vas deferens, constituting the majority of male reproductive tract, are the regions for testosterone production, spermatogenesis, and sperm maturation, storage and discharge, but whether these regions change during aging is unknown. Here, we addressed this by investigating the adult (3-month) and aged (18-month) transcriptomes from seven regions of male mouse reproductive tract：the testis, efferent ductules, initial segment, caput, corpus and cauda epididymis, and vas deferens. We identified various of region-specific genes across different regions of male mouse reproductive tract. Moreover, we identified region-dependent changes of transcripts at an anatomic resolution. This study provides a framework for futher studies on male reproducitve aging.

2022-03-02 | GSE181426 | GEO

Project description:RBFOX splicing factors promote neuronal maturation and axon initial segment assembly

| PRJNA428517 | ENA

RNA sequencing analysis of unilateral orchidectomy, bilateral orchidectomy, and Nell2 knockout initial segment-caput epididymis.

Project description:Purpose: The purpose of this study is to characterize genes whose expressions in the initial segment (IS)-caput epididymis were affected by unilateral orchidectomy (UOD), bilateral orchidectomy (BOD), and Nell2 gene ablation. Methods: The unilateral or bilateral orchidectomy of WT mice was performed from 8 weeks old and continued for 4 weeks until tissue sampling at 12 weeks old. Contralateral epididymis was used as a control for UOD. Sham operation was also done as a control for BOD. Nell2-/- mice and their control WT mice were used wthen they were 14-week-old. IS-caput epididymal mRNA profiles were generated by deep sequencing, in triplicate, using Illumina NovaSeq6000. Results: RNA-seq data identified differentially expressed transcripts. Conclusions: Our results show that the expression of many genes was affected by UOD, BOD, and Nell2 ablation.

2023-11-19 | GSE247764 | GEO

Large-scale analysis of human and mouse RNA-seq datasets to identify undiscovered male reproductive tract-specific genes (mouse)

Project description:The development of a safe and effective, reversible, non-hormonal contraceptive method for men has been an ongoing effort for the past few decades, yet none have reached clinical trials. To advance progress, further identification of novel reproductive tract-specific, potentially druggable protein targets is imperative. In this study, we expand on previous single tissue, single species studies by integrating analysis of a vast number of publicly available human and mouse RNA- seq datasets whose initial published purpose was not focused on identifying male reproductive tract-specific targets. We also incorporate analysis of additional newly acquired human and mouse testis and epididymis samples to increase the number of targets identified. Enclosed in this accession is the RNA-seq sequence data for the 9 new mouse epididymis segment samples we processed for analysis.

2020-07-07 | GSE150853 | GEO

Large-scale analysis of human and mouse RNA-seq datasets to identify undiscovered male reproductive tract-specific genes (human)

Project description:The development of a safe and effective, reversible, non-hormonal contraceptive method for men has been an ongoing effort for the past few decades, yet none have reached clinical trials. To advance progress, further identification of novel reproductive tract-specific, potentially druggable protein targets is imperative. In this study, we expand on previous single tissue, single species studies by integrating analysis of a vast number of publicly available human and mouse RNA-seq datasets whose initial published purpose was not focused on identifying male reproductive tract-specific targets. We also incorporate analysis of additional newly acquired human and mouse testis and epididymis samples to increase the number of targets identified. Enclosed in this accession is the RNA-seq data for the 12 new human testis and epididymis segment samples we processed for analysis.

2020-07-07 | GSE150852 | GEO

The Single-Cell Landscape of Spermioteleosis in Mouse and Pig

Project description:Spermatozoa acquired motility and matured in epididymis after production in testis; however, the characteristics of sperm development between species remain poorly understood. In this study, we constructed a single-cell RNA dataset to investigate spermatogenesis characteristics between mouse and pig. This approach enabled us to analyze cell compositions in both testicular and epididymal tissues, providing insights into changes occurring during meiosis and spermiogenesis in mouse and pig. Additionally, distinct gene expression signatures associated with various spermatogenic cell types were identified. The findings revealed differences in testicular cell clusters between these two species. Furthermore, different regions of the epididymis displayed unique gene expression patterns among its epithelial cells. Notably, regional gene expression patterns were observed within principal cell clusters of mouse epididymis as well. Moreover, we analyzed specific characteristics of differentially expressed genes related to the epididymis in mouse and pig, identifying potential marker genes associated with sperm development and maturation for each species studied.

2024-04-11 | GSE249819 | GEO

Small RNAs are trafficked from the epididymis to developing mammalian sperm

Project description:RNAs present in mature mammalian sperm are delivered to the zygote at fertilization, where they have the potential to affect early development. The biogenesis of the small RNA payload of mature sperm is therefore of great interest, as it may be a target of signaling pathways linking paternal conditions to offspring phenotype. Recent studies have suggested the surprising hypothesis that the small RNA payload carried by mature sperm may include RNAs that were not synthesized during testicular spermatogenesis, but that are instead delivered to sperm during the process of post-testicular maturation in the epididymis. To further test this hypothesis, we characterized small RNA dynamics during testicular and post-testicular germ cell maturation in mice, confirming and extending prior observations that testicular germ cell populations carry extremely low levels of tRNA fragments (tRFs), which only become highly abundant only after sperm have entered the epididymis. We examined the sperm RNA repertoire in greater detail, finding that the majority of 5’ tRNA fragments carry a 2’-3’ cyclic phosphate at their 3’ end, pointing to a role for RNaseA or T-family nucleases in tRNA cleavage in the male reproductive tract. The process of small RNA delivery to sperm can be recapitulated in vitro, as caput epididymosomes deliver small RNAs including tRFs and microRNAs to mature testicular spermatozoa. Finally, to definitively identify the tissue of origin for small RNAs in sperm, we carried out tissue-specific metabolic labeling of RNAs in intact mice, finding that mature sperm carry small RNAs that were originally synthesized in the somatic cells of the epididymis. Taken together, our data demonstrates that soma-germline small RNA transfer occurs in male mammals, most likely via vesicular transport from the epididymis to maturing sperm.

2018-07-15 | GSE112990 | GEO

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