259 The Role of Extracellular Vesicles in Mitigating the Effect of Heat Stress

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 132-133
Author(s):  
Dawit Tesfaye
Keyword(s):  
Oxidative Stress ◽  
Heat Stress ◽  
Extracellular Vesicles ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Biological Fluids ◽  
Biologically Active ◽  
Molecular Signals

Abstract Environmental heat stress negatively affects the fertility of dairy cows by disrupting reproductive processes spanning from follicular development to maternal recognition of pregnancy. Investigation of cellular level responses to stress would contribute to the understanding of the mechanism behind survival responses. Extracellular vesicles (EVs), which carry biologically active signaling molecules, are reported to play a significant role in the cellular response to stress. They are produced by almost all types of cells and abundantly present in various biological fluids including follicular fluid, oviductal fluid, uterine fluids in vivo, and in spent culture media in vitro. Those EV-coupled molecular signals in biological fluids are indicative of the physiological status of the cells of their origin. This has been evidenced by the presence of EV-mediated miRNA signals in follicular fluid associated with the metabolic status of cows. Recent studies revealed the potential role of follicular fluid EVs in carrying molecular signals which can reverse or protect the damage incurred by heat stress in bovine oocytes. In addition to cellular defense responses (activation of HSP70 and HSP90, NRF2 and GRP78 & 94), bovine granulosa cells exposed to heat stress in vitro released EVs enriched with selected mRNA (HSP90 and SOD1) and miRNAs. Among others, miR-1246, miR-374a, and miR-2904 were found to be enriched in EVs released from granulosa cells exposed to thermal stress. Those miRNAs were found to regulate pathways related to heat and endoplasmic reticulum stress responses. The priming of recipient bovine granulosa cells by EVs derived from heat-stressed granulosa cells induced tolerance against recurrent heat stress. Collectively, EV-mediated molecular signals would provide another layer of cell-to-cell communication and deliver protective signals against oxidative stress to recipient cells. This would provide opportunities for future potential application of EVs in tackling oxidative stress-associated fertility problems in humans and animals.

scholarly journals Extracellular vesicles shuttle protective messages against heat stress in bovine granulosa cells

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Samuel Gebremedhn ◽  
Ahmed Gad ◽  
Hoda Samir Aglan ◽  
Jozef Laurincik ◽  
Radek Prochazka ◽  
...  
Keyword(s):  
Heat Stress ◽  
Extracellular Vesicles ◽  
Granulosa Cells ◽  
Beef Cows ◽  
Protective Role ◽  
Bioactive Molecules ◽  
Follicular Cells ◽  
Vitro Model

Abstract Elevated summer temperature is reported to be the leading cause of stress in dairy and beef cows, which negatively affects various reproductive functions. Follicular cells respond to heat stress (HS) by activating the expression of heat shock family proteins (HSPs) and other antioxidants. HS is reported to negatively affect the bi-directional communication between the follicular cells and the oocyte, which is partly mediated by follicular fluid extracellular vesicles (EVs) released from surrounding cells. As carriers of bioactive molecules (DNA, RNA, protein, and lipids), the involvement of EVs in mediating the stress response in follicular cells is not fully understood. Here we used an in vitro model to decipher the cellular and EV-coupled miRNAs of bovine granulosa cells in response to HS. Moreover, the protective role of stress-related EVs against subsequent HS was assessed. For this, bovine granulosa cells from smaller follicles were cultured in vitro and after sub-confluency, cells were either kept at 37 °C or subjected to HS (42 °C). Results showed that granulosa cells exposed to HS increased the accumulation of ROS, total oxidized protein, apoptosis, and the expression of HSPs and antioxidants, while the viability of cells was reduced. Moreover, 14 and 6 miRNAs were differentially expressed in heat-stressed granulosa cells and the corresponding EVs, respectively. Supplementation of stress-related EVs in cultured granulosa cells has induced adaptive response to subsequent HS. However, this potential was not pronounced when the cells were kept under 37 °C. Taking together, EVs generated from granulosa cells exposed to HS has the potential to shuttle bioactive molecules to recipient cells and make them robust to subsequent HS.

scholarly journals Regulation of gene expression in endothelial cells: the role of human follicular fluid

2005 ◽  
Vol 34 (1) ◽  
pp. 37-46 ◽  
Author(s):  
R Gruemmer ◽  
L Klein-Hitpaß ◽  
J Neulen
Keyword(s):  
Gene Expression ◽  
Endothelial Cells ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Regulation Of Gene Expression ◽  
Angiogenic Factors ◽  
Human Follicular Fluid ◽  
Angiopoietin 2

A precise regulation of angiogenesis is a prerequisite for an adequate maturation of ovarian follicles. Despite the production of vascular endothelial growth factor (VEGF) by granulosa cells in antral follicles, angiogenesis is restricted to the theca cell layer. The maturing follicle remains avascular before ovulation, implying regulatory mechanisms which prevent premature follicular vascularization. In order to investigate the role of follicular fluid and of granulosa cells in the regulation of endothelial gene expression, human umbilical vein endothelial cells (HUVECs) were incubated in vitro with media conditioned with human follicular fluid obtained from individual patients undergoing oocyte retrieval for in vitro fertilization procedures or with culture medium conditioned by human granulosa cells respectively. Using microarray technology, the gene expression pattern was compared between untreated monolayers of HUVECs and HUVECs treated either with follicular fluid or with granulosa cell conditioned media. We identified a total of 15 genes that were significantly up-regulated and 11 genes that were significantly down-regulated in endothelial cells treated with follicular fluid at least 2.5-fold in more than 70% of comparisons. Up-regulated genes involved in angiogenesis were the anti-angiogenic factors gro-beta (16.5-fold), angiopoietin-2 (3.9-fold), alpha-2-macroglobulin (24.3-fold) and the pro-angiogenic factors E-selectin (5.3-fold) and vascular cell adhesion molecule-1 (VCAM-1) (4.4-fold), whereas a significant down-regulation of the pro-angiogenic genes fibulin-5 (3.5-fold) and elastin (14.9-fold) could be observed. Culturing of HUVECs with conditioned medium from cultured human luteinized granulosa cells demonstrated a similar regulatory pattern of gene expression for fibulin-5, elastin, gro-beta, and E-selectin. The gene regulation in endothelial cells by follicular fluid could be confirmed by RT-PCR for gro-beta, angiopoietin-2, elastin, fibulin-5, and E-selectin. The present work reveals that compounds secreted by granulosa cells lead to the expression of anti-angiogenic factors on the transcript level in endothelial cells and thus could help to explain the temporal and spatial discrepancy between the high expression of VEGF and the restricted angiogenesis in the preovulatory follicle.

PSX-A-19 Late-Breaking: Potential application of granulosa cell derived extracellular vesicles to mitigate the effects of heat stress in bovine oocytes

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 369-369
Author(s):  
Nico G Menjivar ◽  
Samuel Gebremedhn ◽  
Dawit Tesfaye
Keyword(s):  
Heat Stress ◽  
Thermal Stress ◽  
Granulosa Cell ◽  
Extracellular Vesicles ◽  
Granulosa Cells ◽  
Bystander Effect ◽  
Thermal Conditions ◽  
Developmental Competence ◽  
Cumulus Expansion

Abstract Environmental heat stress negatively affects reproductive efficiency by disrupting follicular development, ultimately compromising gamete competency in cattle. Recently, outlined through the bystander effect, granulosa cell derived extracellular vesicles (EVs) were found to suppress negative effects of recurrent heat stress in recipient bovine granulosa cells. Here, we aimed to assess the effects of supplementing granulosa cell derived EVs during bovine in vitro maturation (IVM) on developmental competence following thermal stress. For this, we modeled a cell culture protocol to generate EVs from bovine granulosa cells subjected to differing ambient temperatures, 38.5°C (body temperature) vs. 42°C (heat stress). At the time of IVM, experimental cumulus oocyte complexes (COCs) were arranged in a 2 x 3 factorial design for temperature (38.5°C or 41°C) versus EV supplementation (normal EVs, stressed EVs and non-supplemented controls) at 20% of the IVM media. Following an initial 8h priming period, half the COCs were subjected to heat shock, the others remained at normal temperature to complete IVM. Results indicate that EV supplementation increased cumulus expansion and the expression of cumulus expansion genes (PTX3, PTGS2 and EGFR). Cleavage rates were increased when supplemented with normal (90.2±1.4%; P = 0.023) or stressed (89.8±2.9%; P = 0.029) EVs, compared to the non-supplemented control (80.5±1.5%) under non-thermal conditions. Similarly, exposure to recurrent thermal stress, cleavage rates were (91±0.9%) and (89±0.6%) when supplemented with normal and stressed EVs respectively, compared to the non-supplemented control (88.5±2.5%). In the absence of exposure to recurrent heat stress, blastocysts rates were (32.4±3.5%) and (31.3±2.9%) when COCs were supplemented with normal and stressed EVs, compared to the control (20.7±4.4%). Blastocysts rates were (23.3±4.7%) and (22.5±3.2%) when COCs were supplemented with normal and stressed EVs, compared to the control (15.5±4.5%) when exposed to recurrent thermal stress. In conclusion, granulosa cell derived EVs have potential to induce oocyte tolerance against recurrent thermal stress.

scholarly journals Effects of Multicycle Gonadotropin-releasing Hormone Antagonist Protocols on Oxidative Stress of Follicular Fluid and Ovarian Granulosa Cells

2020 ◽  
Author(s):  
Yucong Ma ◽  
Zhiming Zhao ◽  
Guimin Hao ◽  
Na Cui ◽  
Yanli Fan ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Gonadotropin Releasing Hormone ◽  
Clinical Pregnancy ◽  
Vitro Fertilization ◽  
Releasing Hormone ◽  
Ovarian Granulosa Cells ◽  
Cycle Group

Abstract Background: Repeated controlled ovarian stimulation (COS) has adverse effects on clinical pregnancy outcomes in in vitro fertilization-embryo transfer (IVF-ET) patients. The effect of repeated antagonist protocols on oxidative stress (OS) in follicular fluid (FF) and ovarian granulosa cells (GC) remains unclear. Objective: To study the effects of repeated multicycle gonadotropin-releasing hormone antagonist (GnRH-ant) protocols on OS markers of FF and ovarian GC. Methods: A total of 145 patients were enrolled and divided into four groups: 1 cycle group (n = 42), 2 cycles group (n = 37), 3 cycles group (n = 45), and 4-5 cycles group (n = 21). The FF and ovarian GC of the patients were collected on the day of last egg retrieval. Levels of 8-hydroxy-2-deoxyguanosine (8-OHdG), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) in FF and ovarian GC were analyzed. Results: With the increase of GnRH-ant protocol cycles, the serum estradiol levels on human chorionic gonadotrophin (hCG) injection day, number of retrieved oocytes, 2PN embryos, and the rates of high-quality embryos, implantation and clinical pregnancy were all significantly decreased, while the gonadotropin (Gn) usage showed an increasing trend. Compared with 1 or 2 cycles, the 8-OHdG and SOD were significantly increased in the 3 to 5 cycles, while the CAT and GSH-Px levels were significantly decreased. Conclusion: Repeated COS with the use of GnRH-ant protocols results in OS and changes the follicle microenvironment of FF and GC, possibly leading to poor IVF outcomes in patients with 3-5 cycles of COS.

scholarly journals ‘Free’ inhibin α subunit is expressed by bovine ovarian theca cells and its knockdown suppresses androgen production

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Mhairi Laird ◽  
Claire Glister ◽  
Warakorn Cheewasopit ◽  
Leanne S. Satchell ◽  
Andrew B. Bicknell ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Follicular Fluid ◽  
Unexpected Finding ◽  
Glycoprotein Hormones ◽  
Positive Role ◽  
Α Subunit ◽  
Theca Cells ◽  
Opposing Effects

AbstractInhibins are ovarian dimeric glycoprotein hormones that suppress pituitary FSH production. They are synthesised by follicular granulosa cells as α plus βA/βB subunits (encoded by INHA, INHBA, INHBB, respectively). Inhibin concentrations are high in follicular fluid (FF) which is also abundant in ‘free’ α subunit, presumed to be of granulosal origin, but its role(s) remains obscure. Here, we report the unexpected finding that bovine theca cells show abundant INHA expression and ‘free’ inhibin α production. Thus, theca cells may contribute significantly to the inhibin α content of FF and peripheral blood. In vitro, knockdown of thecal INHA inhibited INSL3 and CYP17A1 expression and androgen production while INSL3 knockdown reduced INHA and inhibin α secretion. These findings suggest a positive role of thecal inhibin α on androgen production. However, exogenous inhibin α did not raise androgen production. We hypothesised that inhibin α may modulate the opposing effects of BMP and inhibin on androgen production. However, this was not supported experimentally. Furthermore, neither circulating nor intrafollicular androgen concentrations differed between control and inhibin α-immunized heifers, casting further doubt on thecal inhibin α subunit having a significant role in modulating androgen production. Role(s), if any, played by thecal inhibin α remain elusive.

scholarly journals Circulating extracellular vesicles of steroid sensitive nephrotic syndrome patients have higher RAC1 and induce recapitulation of nephrotic syndrome phenotype in podocytes

2021 ◽  
Author(s):  
Fehime Kara Eroglu ◽  
Volkan Yazar ◽  
Ulku Guler ◽  
Muzaffer Yıldırım ◽  
Tugce Yildirim ◽  
...  
Keyword(s):  
Nephrotic Syndrome ◽  
Extracellular Vesicles ◽  
Particle Number ◽  
Biologically Active ◽  
Soluble Mediator ◽  
Steroid Sensitive Nephrotic Syndrome ◽  
Steroid Sensitive

Since previous research suggests a role of a circulating factor in the pathogenesis of steroid-sensitive nephrotic syndrome (SSNS), we speculated that circulating plasma extracellular vesicles (EVs) are a candidate source of such a soluble mediator. Here, we aimed to characterize and try to delineate the effects of these EVs in vitro. Plasma EVs from 20 children with SSNS in relapse and remission, 10 healthy controls and 6 disease controls were obtained by serial ultracentrifugation. Characterization of these EVs was performed by electron microscopy, flow cytometry and western blotting. The major proteins from the plasma EVs were identified via mass spectrometry. A Gene Ontology classification analysis and integuinity pathway analysis were performed on selectively expressed EV proteins during relapse. Immortalized human podocyte culture was used to detect the effects of EVs on podocytes. The protein content and the particle number of plasma EVs were significantly increased during NS relapse. Relapse NS EVs selectively express proteins which involved actin cytoskeleton rearrangement. Among these, the level of RAC-GTP was significantly increased in relapse EVs compared to remission and disease control EVs. Relapse EVs were efficiently internalized by podocytes and induced significantly enhanced motility and albumin permeability. Moreover, relapse EVs induced significantly higher levels of RAC-GTP and phospho p38 (p-p38) and decreased levels of synaptopodin in podocytes. Circulating relapse EVs are biologically active molecules that carry active RAC1 as cargo and induce recapitulation of the nephrotic syndrome phenotype in podocytes in vitro.

scholarly journals The emerging role of the mitochondrial-derived peptide humanin in stress resistance

2012 ◽  
Vol 50 (1) ◽  
pp. R11-R19 ◽  
Author(s):  
Kelvin Yen ◽  
Changhan Lee ◽  
Hemal Mehta ◽  
Pinchas Cohen
Keyword(s):  
Oxidative Stress ◽  
Cardiovascular Disease ◽  
Stress Resistance ◽  
Biologically Active ◽  
Serum Starvation ◽  
Endogenous Role ◽  
Biologically Active Peptide

The discovery of humanin, a novel, mitochondrial-derived peptide, has created a potentially new category of biologically active peptide. As more research unravels the endogenous role of humanin as well as its potential pharmacological use, its role in stress resistance has become clearer. Humanin protects cells from oxidative stress, serum starvation, hypoxia, and other insultsin vitroand also improves cardiovascular disease as well as Alzheimer's diseasein vivo. In this review, we discuss the emerging role of humanin in stress resistance and its proposed mechanism of action.

scholarly journals Extracellular Vesicles: Novel Opportunities to Understand and Detect Neoplastic Diseases

2021 ◽  
pp. 030098582199932
Author(s):  
Laura Bongiovanni ◽  
Anneloes Andriessen ◽  
Marca H. M. Wauben ◽  
Esther N. M. Nolte-’t Hoen ◽  
Alain de Bruin
Keyword(s):  
Extracellular Vesicles ◽  
Biologically Active ◽  
Liquid Biopsies ◽  
Donor Cells ◽  
Recent Developments ◽  
Veterinary Pathology ◽  
Cell Components ◽  
Potential Applications ◽  
Intercellular Communications

With a size range from 30 to 1000 nm, extracellular vesicles (EVs) are one of the smallest cell components able to transport biologically active molecules. They mediate intercellular communications and play a fundamental role in the maintenance of tissue homeostasis and pathogenesis in several types of diseases. In particular, EVs actively contribute to cancer initiation and progression, and there is emerging understanding of their role in creation of the metastatic niche. This fact underlies the recent exponential growth in EV research, which has improved our understanding of their specific roles in disease and their potential applications in diagnosis and therapy. EVs and their biomolecular cargo reflect the state of the diseased donor cells, and can be detected in body fluids and exploited as biomarkers in cancer and other diseases. Relatively few studies have been published on EVs in the veterinary field. This review provides an overview of the features and biology of EVs as well as recent developments in EV research including techniques for isolation and analysis, and will address the way in which the EVs released by diseased tissues can be studied and exploited in the field of veterinary pathology. Uniquely, this review emphasizes the important contribution that pathologists can make to the field of EV research: pathologists can help EV scientists in studying and confirming the role of EVs and their molecular cargo in diseased tissues and as biomarkers in liquid biopsies.

scholarly journals Oxidative Stress as a Possible Target in the Treatment of Toxoplasmosis: Perspectives and Ambiguities

2021 ◽  
Vol 22 (11) ◽  
pp. 5705
Author(s):  
Karolina Szewczyk-Golec ◽  
Marta Pawłowska ◽  
Roland Wesołowski ◽  
Marcin Wróblewski ◽  
Celestyna Mila-Kierzenkowska
Keyword(s):  
Oxidative Stress ◽  
Animal Studies ◽  
Treatment Options ◽  
Natural Antioxidants ◽  
Redox Status ◽  
Host Cells ◽  
Common Disease ◽  
Parasite Viability

Toxoplasma gondii is an apicomplexan parasite causing toxoplasmosis, a common disease, which is most typically asymptomatic. However, toxoplasmosis can be severe and even fatal in immunocompromised patients and fetuses. Available treatment options are limited, so there is a strong impetus to develop novel therapeutics. This review focuses on the role of oxidative stress in the pathophysiology and treatment of T. gondii infection. Chemical compounds that modify redox status can reduce the parasite viability and thus be potential anti-Toxoplasma drugs. On the other hand, oxidative stress caused by the activation of the inflammatory response may have some deleterious consequences in host cells. In this respect, the potential use of natural antioxidants is worth considering, including melatonin and some vitamins, as possible novel anti-Toxoplasma therapeutics. Results of in vitro and animal studies are promising. However, supplementation with some antioxidants was found to promote the increase in parasitemia, and the disease was then characterized by a milder course. Undoubtedly, research in this area may have a significant impact on the future prospects of toxoplasmosis therapy.

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