scholarly journals Ferroptosis induces membrane blebbing in placental trophoblasts

2021 ◽  
pp. jcs.255737
Author(s):  
Kazuhiro Kajiwara ◽  
Ofer Beharier ◽  
Choon-Peng Chng ◽  
Julie P. Goff ◽  
Yingshi Ouyang ◽  
...  
Keyword(s):  
Cell Death ◽  
Plasma Membrane ◽  
Molecular Modeling ◽  
Cell Membrane ◽  
Glutathione Peroxidase ◽  
Oxidative Damage ◽  
Membrane Lipids ◽  
Target Cells ◽  
Cytoplasmic Organelle ◽  
Membrane Blebbing

Ferroptosis is a regulated, non-apoptotic form of cell death, characterized by hydroxy-peroxidation of discrete phospholipid hydroperoxides, particularly hydroperoxyl (Hp)- forms of arachidonoyl- and adrenoyl-phosphatidylethanolamine, with a downstream cascade of oxidative damage to membrane lipids, proteins, and DNA, culminating in cell death. We recently showed that human trophoblasts are particularly sensitive to ferroptosis, caused by depletion or inhibition of glutathione peroxidase 4 (GPX4) or the lipase PLA2G6. Here, we show that trophoblastic ferroptosis is accompanied by a dramatic change in trophoblast plasma membrane, with macro-blebbing and vesiculation. Immunofluorescence revealed that ferroptotic cell-derived blebs stained positive for F-actin, but negative for cytoplasmic organelle markers. Transfer of conditioned medium that contained detached macrovesicles or co-culture with blebbing cells did not stimulate ferroptosis in target cells. Molecular modeling showed that the presence of Hp- phosphatidylethanolamine in the cell membrane promoted its stretchability. Together, our data establish that membrane macro-blebbing is characteristic of trophoblast ferroptosis and can serve as a useful marker of this process. Whether or not these blebs are physiologically functional remains to be established.

Lateral mobility of plasma membrane lipids in bull spermatozoa: heterogeneity between surface domains and rigidification following cell death

1997 ◽  
Vol 110 (9) ◽  
pp. 1041-1050 ◽  
Author(s):  
S. Ladha ◽  
P.S. James ◽  
D.C. Clark ◽  
E.A. Howes ◽  
R. Jones
Keyword(s):  
Cell Death ◽  
Plasma Membrane ◽  
Membrane Lipids ◽  
Surface Membrane ◽  
Fold Increase ◽  
Membrane Lipid ◽  
Differentiated Cells ◽  
Immobile Phase ◽  
Lipid Diffusion ◽  
Surface Domains

Compartmentalization of surface membrane antigens into discrete regions or domains is a characteristic feature of differentiated cells. In mammalian spermatozoa at least 5 surface domains are known, implying the presence of barriers or boundaries within the plasma membrane. Using the technique of fluorescence recovery after photobleaching (FRAP) to measure diffusibility of fluorescent lipid analogues 1,1′-dihexadecyl-3,3,3′3′-tetramethylindocarbocyanine (DiIC[16]) and 5-(N-octa-decanoyl) aminofluorescein (ODAF), we have investigated lipid topology and dynamics in the plasma membrane of ejaculated bull spermatozoa. Contrary to reports in the literature, we have found that DiIC(16) stains only dead or damaged spermatozoa whereas ODAF intercalates into the plasma membrane of both live and dead cells, each type showing a distinctive staining pattern. FRAP analysis with ODAF revealed that diffusion coefficients on live spermatozoa are significantly faster on the acrosome and postacrosome (29.3x10(−9) cm2/second) than on the midpiece and principal piece (11.8x10(−9) cm2/second). Recovery (R) is >90% in all domains. ODAF diffusion also shows regionalized temperature-sensitivity with a 4-fold increase over the sperm head and a 1.8-fold increase on the tail between 20 degrees C and 37 degrees C. Remarkably, dead or permeabilized spermatozoa rapidly develop a large immobile phase (R<25%) over the whole plasma membrane. This rigidification is temperature insensitive and irreversible suggesting major changes in the physical state of membrane lipids. It is concluded that lipid diffusion in the plasma membrane of live bull spermatozoa is rapid and varies significantly between surface domains. Following permeabilization or cell death, however, a large immobile phase develops indicating substantial changes in membrane lipid disposition.

scholarly journals Dexamethasone blocks antioestrogen- and oxidant-induced death of pituitary tumour cells

2001 ◽  
Vol 169 (2) ◽  
pp. 249-261 ◽  
Author(s):  
CJ Newton ◽  
D Bilko ◽  
S Pappa ◽  
SL Atkin
Keyword(s):  
Cell Death ◽  
Cell Membrane ◽  
Tumour Cell ◽  
Time Course ◽  
Apoptotic Cell ◽  
Cell Types ◽  
Pituitary Tumour ◽  
Tumour Cell Line ◽  
Membrane Function ◽  
Membrane Blebbing

The oestrogen receptor is fundamental to the growth and survival of the rat pituitary tumour cell line, GH(3). Our previous studies have shown that antioestrogens such as RU 58668 and ZM 182780 will reduce the rate of cell division and also induce cell death. Death of these cells in response to antioestrogen treatment appears to be due to a heightened sensitivity to reactive oxygen species (ROS). As part of a study to determine the cross-talk between steroid receptor systems in these cells, we have observed that the glucocorticoid, dexamethasone (Dex), inhibits antioestrogen-induced cell death. Cell death induced by H(2)O(2) is enhanced by ZM 182780 and this effect is also blocked by Dex. As apoptotic cell death in a number of systems involves an early loss of mitochondrial membrane potential (DeltaPsi(m)), we have performed detailed studies on the time-course of DeltaPsi(m) loss in relation to the loss in cell membrane function. These studies have indicated that a loss of DeltaPsi(m) parallels a loss of cell membrane function - this is more characteristic of necrosis than of apoptosis. From microscopic observations of these cells in response to H(2)O(2), it has been noted that early cell membrane blebbing, induced by H(2)O(2), is blocked in the presence of ZM 182780. Cell membrane blebbing can precede necrosis as well as apoptosis and it is thought to involve cytoskeletal changes, for which localised glycolytic reactions provide ATP. These observations, together with those showing that removal of glucose, but not inhibition of mitochondrial function, enhances ROS-induced cell death, prompted studies on the glycolytic pathway. As a strong candidate mechanism, it would appear that, via an effect on one of the rate-limiting glycolytic enzymes, glyceraldehyde-3-phosphate dehydrogenase, Dex is able to overcome the antioestrogen-enhanced loss of glycolytic function following exposure of cells to ROS. This report contributes to the growing body of evidence showing that glucocorticoids provide a survival advantage to both normal and tumour cell types.

Toxicity Mechanism of Gadolinium Oxide Nanoparticles and Gadolinium Ions in Human Breast Cancer Cells

2019 ◽  
Vol 20 (11) ◽  
pp. 907-917 ◽  
Author(s):  
Mohd Javed Akhtar ◽  
Maqusood Ahamed ◽  
Hisham Alhadlaq ◽  
Salman Alrokayan
Keyword(s):  
Breast Cancer ◽  
Cell Death ◽  
Cell Membrane ◽  
Oxidative Damage ◽  
Human Breast Cancer ◽  
Membrane Damage ◽  
Gadolinium Oxide ◽  
Human Breast ◽  
Mechanisms Of Toxicity

Background: Due to the potential advantages of Gadolinium Nanoparticles (NPs) over gadolinium elements, gadolinium based NPs are currently being explored in the field of MRI. Either in elemental form or nanoparticulate form, gadolinium toxicity is believed to occur due to the deposition of gadolinium ion (designated as Gd3+ ion or simply G ion). Objective: There is a serious lack of literature on the mechanisms of toxicity caused by either gadolinium-based NPs or ions. Breast cancer tumors are often subjected to MRIs, therefore, human breast cancer (MCF-7) cells could serve as an appropriate in vitro model for the study of Gadolinium Oxide (GO) NP and G ion. Methods: Cytotoxicity and oxidative damage was determined by quantifying cell viability, cell membrane damage, and Reactive Oxygen Species (ROS). Intracellular Glutathione (GSH) was measured along with cellular Total Antioxidant Capacity (TAC). Autophagy was determined by using Monodansylcadaverine (MDC) and Lysotracker Red (LTR) dyes in tandem. Mitochondrial Membrane Potential (MMP) was measured by JC-1 fluorescence. Physicochemical properties of GO NPs were characterized by field emission transmission electron microscopy, X-ray diffraction, and energy dispersive spectrum. Results: A time- and concentration-dependent toxicity and oxidative damage was observed due to GO NPs and G ions. Bax/Bcl2 ratios, FITC-7AAD double staining, and cell membrane blebbing in phase-contrast images all suggested different modes of cell death induced by NPs and ions. Conclusion: In summary, cell death induced by GO NPs with high aspect ratio favored apoptosis-independent cell death, whereas G ions favored apoptosis-dependent cell death.

scholarly journals Direct cleavage of ROCK II by granzyme B induces target cell membrane blebbing in a caspase-independent manner

2005 ◽  
Vol 201 (3) ◽  
pp. 465-471 ◽  
Author(s):  
Michael Sebbagh ◽  
Jocelyne Hamelin ◽  
Jacques Bertoglio ◽  
Eric Solary ◽  
Jacqueline Bréard
Keyword(s):  
Cell Death ◽  
Granzyme B ◽  
Coiled Coil ◽  
Target Cells ◽  
Caspase Activation ◽  
Cytotoxic Lymphocyte ◽  
Independent Manner ◽  
Major Function ◽  
Membrane Blebbing ◽  
Induced Apoptosis

Caspase activation in target cells is a major function of granzyme B (grB) during cytotoxic lymphocyte granule-induced apoptosis. grB-mediated cell death can occur in the absence of active caspases, and the molecular targets responsible for this additional pathway remain poorly defined. Apoptotic plasma membrane blebbing is caspase independent during granule exocytosis–mediated cell death, whereas in other instances, this event is a consequence of the cleavage by caspases of the Rho effector, Rho-associated coiled coil–containing protein kinase (ROCK) I. We show here that grB directly cleaves ROCK II, a ROCK family member encoded by a separate gene and closely related to ROCK I, and this causes constitutive kinase activity and bleb formation. For the first time, two proteins of the same family are found to be specifically cleaved by either a caspase or grB, thus defining two independent pathways with similar phenotypic consequences in the cells. During granule-induced cell death, ROCK II cleavage by grB would overcome, for this apoptotic feature, the consequences of deficient caspase activation that may occur in virus-infected or malignant target cells.

scholarly journals Bcl-xL overexpression attenuates glutathione depletion in FL5.12 cells following interleukin-3 withdrawal

1997 ◽  
Vol 325 (2) ◽  
pp. 315-319 ◽  
Author(s):  
Heidi K. BOJES ◽  
Kaushik DATTA ◽  
Jie XU ◽  
Anita CHIN ◽  
Phil SIMONIAN ◽  
...  
Keyword(s):  
Cell Death ◽  
Superoxide Dismutase ◽  
Glutathione Peroxidase ◽  
Oxidative Damage ◽  
Glutathione Reductase ◽  
Cell Line ◽  
Incubation Period ◽  
Glutathione Depletion ◽  
Rapid Loss ◽  
Induction Of Apoptosis

Bcl-xL and bax are bcl-2-related genes whose protein products either inhibit or promote apoptosis. Oxidative damage, including the loss of glutathione, has been implicated in the induction of apoptosis. The ability of the Bcl proteins to affect GSH was assessed in control, bax- and bcl-xL-transfected FL5.12 cells [an interleukin (IL)-3-dependent murine prolymphocytic cell line]. Overall levels of GSH were approximately the same in control and bcl-xL transfectants during the 6 h incubation period, although levels increased in bcl-xL transfectants 24 h after replating. GSH in cells overexpressing bax was reduced by ∼ 36%. There were no consistent differences between these cell lines in the activities of superoxide dismutase, catalase, glutathione peroxidase or glutathione reductase. Following IL-3 withdrawal, a condition known to cause apoptosis in these cells, a rapid loss of intracellular GSH occurred in control and bax transfectants, which preceded the onset of apoptosis. GSH depletion could not be attributed to intracellular oxidation but rather seemed to occur due to a translocation out of the cell. Cells overexpressing bcl-xL did not lose significant amounts of GSH upon withdrawal of IL-3, and no apoptosis was evident. These results suggest a possible role for GSH in the mechanism by which bcl-xL prevents cell death.

scholarly journals Cationic Sites on Granzyme B Contribute to Cytotoxicity by Promoting Its Uptake into Target Cells

2005 ◽  
Vol 25 (17) ◽  
pp. 7854-7867 ◽  
Author(s):  
Catherina H. Bird ◽  
Jiuru Sun ◽  
Kheng Ung ◽  
Diana Karambalis ◽  
James C. Whisstock ◽  
...  
Keyword(s):  
Cell Death ◽  
Plasma Membrane ◽  
Cell Surface ◽  
Chondroitin Sulfate ◽  
Granzyme B ◽  
Target Cells ◽  
Adsorption Model ◽  
Cytotoxic Lymphocyte ◽  
Intracellular Compartment ◽  
Exchange Adsorption

ABSTRACT Granzyme B (GrB) is a key effector of cytotoxic lymphocyte-mediated cell death. It is delivered to target cells bound to the proteoglycan serglycin, but how it crosses the plasma membrane and accesses substrates in the cytoplasm is poorly understood. Here we identify two cationic sequences on GrB that facilitate its binding and uptake. Mutation of cationic sequence 1 (cs1) prevents accumulation of GrB in a distinctive intracellular compartment and reduces cytotoxicity 20-fold. Mutation of cs2 reduces accumulation in this intracellular compartment and cytotoxicity two- to threefold. We also show that GrB-mediated cytotoxicity is abrogated by heparin and that target cells deficient in cell surface sulfate or glycosaminoglycans resist GrB. However, heparin does not completely prevent GrB internalization and chondroitin 4-sulfate does not inhibit cytotoxicity, suggesting that glycosaminoglycans are not essential GrB receptors. We propose that GrB enters cells by nonselective adsorptive pinocytosis, exchanging from chondroitin sulfate on serglycin to anionic components of the cell surface. In this electrostatic “exchange-adsorption” model, cs1 and cs2 participate in binding of GrB to the cell surface, thereby promoting its uptake and eventual release into the cytoplasm.

scholarly journals Plasma Membrane Repair: A Central Process for Maintaining Cellular Homeostasis

Physiology ◽  
2015 ◽  
Vol 30 (6) ◽  
pp. 438-448 ◽  
Author(s):  
Alisa D. Blazek ◽  
Brian J. Paleo ◽  
Noah Weisleder
Keyword(s):  
Cell Death ◽  
Plasma Membrane ◽  
Cell Membrane ◽  
Cellular Response ◽  
Cellular Functions ◽  
Membrane Repair ◽  
Central Process ◽  
Plasma Membrane Repair ◽  
Molecular Machinery ◽  
Membrane Resealing

Plasma membrane repair is a conserved cellular response mediating active resealing of membrane disruptions to maintain homeostasis and prevent cell death and progression of multiple diseases. Cell membrane repair repurposes mechanisms from various cellular functions, including vesicle trafficking, exocytosis, and endocytosis, to mend the broken membrane. Recent studies increased our understanding of membrane repair by establishing the molecular machinery contributing to membrane resealing. Here, we review some of the key proteins linked to cell membrane repair.

scholarly journals Targeting Scavenger Receptor Type B1 In Cholesterol-Addicted Lymphomas Abolishes Glutathione Peroxidase 4 and Results in Ferroptosis

2020 ◽  
Author(s):  
Jonathan S. Rink ◽  
Adam Yuh Lin ◽  
Kaylin M. McMahon ◽  
Andrea E. Calvert ◽  
Shuo Yang ◽  
...  
Keyword(s):  
Cell Death ◽  
Glutathione Peroxidase ◽  
Cholesterol Synthesis ◽  
De Novo ◽  
Metabolic Response ◽  
Membrane Lipids ◽  
Scavenger Receptor ◽  
Receptor Type ◽  
Lymphoma Cells ◽  
Cholesterol Uptake

ABSTRACTNormal human cells can either synthesize or uptake cholesterol from lipoproteins to meet their metabolic requirements. Some malignant cells absolutely require cholesterol uptake from lipoproteins for survival because de novo cholesterol synthesis genes are transcriptionally silent or mutated. Recent data suggest that lymphoma cells dependent upon lipoprotein-mediated cholesterol uptake are also dependent on the expression of the lipid hydroperoxidase enzyme glutathione peroxidase 4 (GPX4) to prevent cell death by ferroptosis. Ferroptosis is an oxygen-and iron-dependent cell death mechanism that results from the accumulation of oxidized lipids in cell membranes. To study mechanisms linking cholesterol uptake with ferroptosis, we employed lymphoma cell lines known to be sensitive to cholesterol uptake depletion and treated them with high-density lipoprotein-like (HDL) nanoparticles (HDL NPs). HDL NPs are a cholesterol-poor ligand of the receptor for cholesterol-rich HDL, scavenger receptor type B-1 (SCARB1). Our data reveal that HDL NP treatment activates a compensatory metabolic response in treated cells favoring de novo cholesterol synthesis, which is accompanied by reduced expression of GPX4. As a result, accumulation of oxidized membrane lipids leads to cell death through a mechanism consistent with ferroptosis. Furthermore, ferroptosis was validated in vivo after systemic administration of HDL NPs in mouse lymphoma xenografts and in primary samples obtained from patients with lymphoma. In summary, targeting SCARB1 with HDL NPs in cholesterol uptake addicted lymphoma cells abolishes GPX4 and cancer cell death ensues through a mechanism consistent with ferroptosis.

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