Primary cilia are shear stress calcium-responsive mechanosensors in the epididymis
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PROVIDER: PRJNA1124044 | ENA |
REPOSITORIES: ENA
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Project description:The epithelium of the epididymis proliferates and differentiates until puberty, to become fully functional when the first wave of spermatozoa initiates its post-testicular maturation process. While the tension exerted by the testicular-derived fluid has been proposed to stimulate the proliferation/differentiation of the epithelium through physical cues, the mechanism involved has never been identified. Here we assessed whether the responsiveness of the epididymal cells to luminal shear stress depends on functional primary cilia. Combining in silico fluidic modeling with in vitro microfluidic strategies, we subjected immortalized distal caput epididymis principal cells (DC2) to a static or fluid shear stress condition. RNA sequencing studies identified 100 genes as being overexpressed in response to shear stress (Fold change >1.5, FDR< 0.05), including Early growth response (EGR) 2/3, Cellular communication network factor (CCN) 1/2, and Fos, also found induced in flow-stimulated renal cells. Enrichment of transcriptional factors involved in proliferative signaling pathways was consistent with the proliferative changes observed in DC2 cells under shear stress conditions. Of interest, DEGs particularly relevant to epididymal physiology and sperm maturation, including Serpine1 and Adamts1, were identified as secreted factors interacting with the maturing spermatozoa. While shear stress triggered a rapid increase of intracellular Ca2+ in DC2 cells, this response was abrogated following the impairment of primary ciliogenesis through pharmacological and siRNA approaches. Embracing physical and biological concepts, our findings identify primary cilia as a fluid-mechanosensor that may contribute to epididymal cell development and proliferation, a requisite to sperm maturation
2025-01-22 | GSE269872 | GEO
Project description:Analysis of mechano-regulation of mesenchymal stem cell gene expression level. The hypothesis tested in the present study was that laminar shear stress influences the cytoskeleton arrangement and amount and then triggers seriers of cell metabolism and cellular functions. Results provide important information of the response of mesenchymal stem cell to laminar shear stress, such as specific mechano-responsive genes, up- or down-regulated specific anabolic/catabolic cellular functions.
2018-12-31 | GSE109180 | GEO
Project description:Biomechanical forces such as wall shear stress induced by flowing blood influence the behavior of vascular cells and have been shown to be crucial in the onset of vascular disease. Primary cilia are increasingly recognized as important biological sensors of low WSS. The goal of the study was to investigate gene expression profile of endothelial cell under low flow (2 dynes/cm2) as compared to physiological flow (30 dynes/cm2) in presence or absence of primary cilia.
2022-05-25 | GSE139580 | GEO
Project description:Cardiopulmonary bypass (CPB), the mechanical support required during the majority of cardiac surgeries, causes significant systemic inflammation and multi-organ dysfunction that is especially severe in neonatal patients. Currently, the limited understanding of the molecular mechanisms that underlie CPB-associated inflammation presents a significant barrier to the development of therapeutic interventions. To address this knowledge gap, we performed mRNA sequencing of total circulating leukocytes from neonatal patients undergoing CPB. The transcriptional signatures across 7 time points ranging from pre-CPB, during CPB, to 24 hours post-CPB were found to be dominated by myeloid cells, particularly monocytes. INTERLEUKIN-8 (IL8) and TUMOR NECROSIS FACTOR-α (TNFα) were two inflammatory cytokines specifically and robustly upregulated in the leukocytes from both CPB patients and piglets exposed to CPB. To delineate the mechanism how CPB activates cytokine production, we exposed THP-1 cells, a human monocyte cell line, in vitro to CPB-like conditions including artificial surfaces, high shear stress and cooling/rewarming. The in vitro CPB data recapitulated the in vivo CPB condition in that IL8 and TNFα were strongly upregulated. Shear stress was the major driver of cytokine expression as compared to the temperature or plastic exposure. Extracellular calcium influx was found essential for the shear-stress mediated upregulation of cytokines via the MEK/ERK/AP-1 and Calcineurin/NFAT pathways. After being exposed to CPB conditions, a subpopulation of THP-1 cells died by apoptosis and TNFα-mediated necroptosis, the latter form of which potentially contribute to the post-CPB tissue injuries. Together, our study is the first to elucidate the shear-stress modulated molecular mechanisms leading to systemic inflammation in pediatric CPB and the link between inflammation and organ damage. Calcium signaling pathways and TNFα signaling are novel targets for therapeutic development to improve clinical outcomes. To our knowledge, these are also the first data to implicate necroptosis in CPB and show that shear stress can activate necroptosis.
2020-12-06 | GSE143780 | GEO
Project description:Background: Whereas cilia damage and reduced cilia beat frequency have been implicated as causative of reduced mucociliary clearance in smokers, theoretically mucociliary clearance could also be affected by cilia length. Based on models of mucociliary clearance predicting cilia length must exceed the 6 -7 μm airway surface fluid depth to generate force in the mucus layer, we hypothesized cilia height may be decreased in airway epithelium of normal smokers compared to nonsmokers. Methodology/Principal Findings: Cilia length in normal nonsmokers and smokers was evaluated in aldehyde-fixed, paraffin-embedded endobronchial biopsies, and air-dried and hydrated samples brushed from human airway epithelium via fiberoptic bronchoscopy. In 28 endobronchial biopsies, healthy smoker cilia length was reduced 15% compared to nonsmokers (p<0.05). In 47 air-dried samples of airway epithelial cells, smoker cilia length was reduced 13% compared to nonsmokers (p<0.0001). Analysis of the length of individual, detached cilia in 17 samples, smoker cilia length was reduced 9% compared to nonsmokers (p<0.05). Finally, in 16 fully hydrated, unfixed samples, smoker cilia length was reduced 7% compared to nonsmokers (p<0.05). Conclusions/significance: Models predict that a reduction in cilia length would reduce mucociliary clearance, suggesting that smoking-associated shorter airway epithelial cilia plays a significant role in the pathogenesis of smoking-induced lung disease.
2012-12-13 | E-GEOD-16696 | biostudies-arrayexpress
Project description:Identification of atheroprone shear stress responsive regulatory elements in endothelial cells
| PRJNA477847 | ENA
Project description:Microarray analysis was used to identify the calcium-responsive genes dependent on CrzA in the filamentous fungus Aspergillus nidulans. In order to identify such genes, we conducted the two types of experiment. One was a comparison between wild type with calcium treatment and wild type without calcium treatment. Another was a comparison between wild type with calcium treatment and crzA mutant with calcium treatment. From a comparison between the results of these experiments, we could identify the genes whose expression was induced or repressed in response to calcium in a manner dependent on CrzA. KEY WORD; Aspergillus nidulans, calcium response, crzA
2008-10-03 | GSE12953 | GEO
Project description:Primary cilia are solitary, generally non-motile, hair-like protrusions that extend from the surface of cells between cell divisions. Their antenna-like structure leads naturally to the assumption that they sense the surrounding environment, the most common hypothesis being sensation of mechanical force through calcium-permeable ion channels within the cilium. This Ca(2+)-responsive mechanosensor hypothesis for primary cilia has been invoked to explain a large range of biological responses, from control of left-right axis determination in embryonic development to adult progression of polycystic kidney disease and some cancers. Here we report the complete lack of mechanically induced calcium increases in primary cilia, in tissues upon which this hypothesis has been based. We developed a transgenic mouse, Arl13b-mCherry-GECO1.2, expressing a ratiometric genetically encoded calcium indicator in all primary cilia. We then measured responses to flow in primary cilia of cultured kidney epithelial cells, kidney thick ascending tubules, crown cells of the embryonic node, kinocilia of inner ear hair cells, and several cell lines. Cilia-specific Ca(2+) influxes were not observed in physiological or even highly supraphysiological levels of fluid flow. We conclude that mechanosensation, if it originates in primary cilia, is not via calcium signalling.
| S-EPMC4851444 | biostudies-literature
Project description:While variations in gene transcription networks in models of atherosclerosis have been reported, the underlying changes in the chromatin landscape induced by pro-atherogenic stimuli remain elusive. In the present study, we report changes in chromatin regulatory elements that mediate transcriptional control upon the application of oscillatory shear stress of ±3 dyn/cm2 in primary cultured human umbilical vein endothelial cells (HUVECs) at 6 h time point of oscillatory shear stress stimulation compared to static condition.
2019-03-04 | GSE116241 | GEO
Project description:The role of estrogen and testosterone in the regulation of gene expression in the proximal reproductive tract is not completely understood. To address this question, mice were treated with testosterone or estradiol and RNA from the efferent ducts and caput epididymis was processed and hybridized to Affymetrix MOE 430 2.0 microarrays. Analysis of array output identified probe sets in each tissue with altered levels in hormone treated versus control animals. Hormone treatment efficacy was confirmed by determination of serum hormone levels pre- and post-treatment and observed changes in transcript levels of previously reported hormone-responsive genes. Tissue-specific hormone sensitivity was observed with 2867 and 3197 probe sets changing significantly in the efferent ducts after estrogen and testosterone treatment, respectively. In the caput epididymis, 117 and 268 probe sets changed after estrogen and testosterone treatment, respectively, demonstrating a greater response to hormone in the efferent ducts than the caput epididymis. Transcripts sharing similar profiles in the intact and hormone-treated animals compared with castrated controls were also identified. Ontological analysis of probe sets revealed a significant number of hormone-regulated transcripts encode proteins associated with lipid metabolism, transcription and steroid metabolism in both tissues. Real-time RT-PCR was employed to confirm array data and investigate other potential hormone-responsive regulators of proximal reproductive tract function. The results of this work reveal previously unknown responses to estrogen in the caput epididymis and to testosterone in the efferent ducts as well as tissue specific hormone sensitivity in the proximal reproductive tract. Adult animals were castrated or sham-castrated, allowed to recover for 14 days, and then treated with 0.015 mg estradiol (castrated), 0.015 mg testosterone propionate (castrated), or vehicle (castrated and sham-castrated as biological controls) in duplicate. Efferent duct and caput epididymis was collected from each sample and analyzed. Duplicates are included in the provided data and numbered 1 or 2 for each treatment regimen.
2009-12-30 | E-GEOD-17553 | biostudies-arrayexpress