scholarly journals α-Fodrin in Cytoskeletal Organization and the Activity of Certain Key Microtubule Kinesins

Genes ◽  
2021 ◽  
Vol 12 (5) ◽  
pp. 750
Author(s):  
Jamuna S. Sreeja ◽  
Athira Jyothy ◽  
Suparna Sengupta
Keyword(s):  
Cell Cycle ◽  
Plasma Membrane ◽  
Cell Adhesion ◽  
Ex Vivo ◽  
Membrane Integrity ◽  
Cytoskeletal Proteins ◽  
Cellular Mechanisms ◽  
Cytoskeletal Organization ◽  
Plasma Membrane Integrity

Cortical cytoskeletal proteins are significant in controlling various cellular mechanisms such as migration, cell adhesion, intercellular attachment, cellular signaling, exo- and endocytosis and plasma membrane integrity, stability and flexibility. Our earlier studies involving in vitro and ex vivo approaches led us to identify certain undiscovered characteristics of α-fodrin, a prominent cortical protein. The conventional functions attributed to this protein mainly support the plasma membrane. In the present study, we utilized a global protein expression analysis approach to detect underexplored functions of this protein. We report that downregulation of α-fodrin in glioblastoma cells, U-251 MG, results in upregulation of genes affecting the regulation of the cytoskeleton, cell cycle and apoptosis. Interestingly, certain key microtubule kinesins such as KIF23, KIF2B and KIF3C are downregulated upon α-fodrin depletion, as validated by real-time PCR studies.

scholarly journals Effects of cryopreservation on sperm quality, nuclear DNA integrity, in vitro fertilization, and in vitro embryo development in the mouse

Reproduction ◽  
2007 ◽  
Vol 133 (3) ◽  
pp. 585-595 ◽  
Author(s):  
Cengiz Yildiz ◽  
Palma Ottaviani ◽  
Napoleon Law ◽  
Renise Ayearst ◽  
Ling Liu ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Embryo Development ◽  
Membrane Integrity ◽  
Fertilization Rate ◽  
Development Rate ◽  
Dna Integrity ◽  
Plasma Membrane Integrity ◽  
Mouse Sperm ◽  
Progressive Motility

Efficient freezing, archiving, and thawing of sperm are essential techniques to support large scale research programs using mouse models of human disease. The purpose of this study was to investigate the effects of variable combinations and concentrations of cryoprotectants on sperm-assessment parameters of frozen–thawed mouse sperm in order to optimize cryopreservation protocols. Sperm was frozen using combinations of 3% skim milk + 0.2 or 0.3 M nonpermeating raffinose with either permeating glucose, fructose, propylene glycol, ethylene glycol, glycerol, or sodium pyruvate in CD-1, C3FeB6F1/J, B6129SF1, C57BL/6NCrIBR, 129S/SvPaslco, and DBA/2NCrIBR mice. Sperm-assessment parameters included progressive motility, plasma membrane integrity (SYBR-14 + PI),in vitrofertilization rate, andin vitroembryo development rate to blastocyst. DNA content analysis of sperm was measured by the sperm chromatin structure assay (SCSA). 0.3 M raffinose with 0.1 M fructose significantly improved post-thaw sperm-assessment parameters for CD-1, C3B6F1, B6129SF1 mice (P< 0.05–0.01), whereas 0.2 M raffinose with 0.1 M glycerol or 0.1 M fructose enhanced sperm assessment values for C57BL/6 and 129S mice (P< 0.01), compared to 0.3 M raffinose alone. DNA fragmentation during cryopreservation was significantly increased in all strains evaluated when compared with fresh control sperm in a strain-dependent manner (P< 0.01). Supplementation with permeating glycerol or fructose to the cryoprotectant (CPA) solution showed a significant protective effect to DNA integrity when cryopreserving sperm from C57BL/6 and 129S mice. Damage to sperm DNA significantly decreased the rate ofin vitroembryo development to blastocyst in C57BL/6 mice. The type of monosaccharide sugar or polyols, CPA molarity, and combination of permeating and nonpermeating cryoprotectant are significant factors for improving progressive motility, plasma membrane integrity, DNA integrity,in vitrofertilization rate, andin vitroembryo development rate to blastocyst in cryopreserved mouse sperm.

scholarly journals Polystyrene nanoplastics and microplastics can act as Trojan horse carriers of benzo(a)pyrene to mussel hemocytes in vitro

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alberto Katsumiti ◽  
María Paula Losada-Carrillo ◽  
Marta Barros ◽  
Miren P. Cajaraville
Keyword(s):  
Plasma Membrane ◽  
Cell Viability ◽  
Phagocytic Activity ◽  
Membrane Integrity ◽  
Aquatic Organisms ◽  
Particle Aggregation ◽  
Ros Production ◽  
Plasma Membrane Integrity ◽  
Intracellular Fate

AbstractIn this work we studied the ability of polystyrene (PS) nanoplastics (NPs) and microplastics (MPs) to transfer benzo(a)pyrene (BaP) to mussel hemocytes and to produce toxic effects in vitro. For this, intracellular fate and toxicity of PS NPs (0.05 μm) and MPs (0.5 and 4.5 μm) alone or with BaP and of BaP alone were assessed. Particles of 0.05 and 0.5 µm largely aggregated in the exposure medium whereas presence of BaP reduced particle aggregation. Cells internalized PS NPs and MPs alone or with BaP and these were found inside and outside lysosomes, depending on their size. PS particles alone or with BaP were cytotoxic to hemocytes only at the highest concentrations tested. The same was true for most sublethal endpoints except for increased phagocytic activity provoked by NPs and 0.5 μm MPs at lower concentrations. Plastic particles appeared to be the main drivers for reduced plasma membrane integrity and increased phagocytic and lysosomal activities whereas BaP appeared to contribute more to reduced cell viability and phagocytosis and increased ROS production and genotoxicity. Overall, PS NPs and MPs can act as carriers of BaP to mussel hemocytes, rising concerns about risks plastics associated to pollutants may pose to aquatic organisms.

scholarly journals Antisperm antibodies disrupt plasma membrane integrity and inhibit tyrosine phosphorylation in human spermatozoa

2018 ◽  
Vol 27 (1) ◽  
pp. 3-11 ◽  
Author(s):  
Dwi A. Pujianto ◽  
Hajizah Hajizah ◽  
Indra G. Mansur ◽  
Amarudin Amarudin
Keyword(s):  
Plasma Membrane ◽  
Tyrosine Phosphorylation ◽  
Membrane Integrity ◽  
In Vitro Study ◽  
Dependent Manner ◽  
Sperm Viability ◽  
Plasma Membrane Integrity ◽  
Infertile Women ◽  
The Status

Background: The etiology of unexplained infertility has not been fully understood. This study aimed to determine the effect of antisperm antibody (ASA) from infertile women on viability, motility, plasma membrane integrity, and status of tyrosine phosphorylation in the human spermatozoa.Methods: An experimental in vitro study was conducted at the Department of Biology, Faculty of Medicine, Universitas Indonesia from February to November 2014. Spermatozoa from normal fertile donors was incubated with serum containing ASA from infertile women at several dilutions (1/1000, 1/100, 1/10, and without dilution) for 1 and 2 hours. The plasma membrane integrity was assessed with hypoosmotic swelling (HOS) test, whereas the status of tyrosine phosphorylation was analyzed using Western immunoblotting and immunocytochemistry.Results: After 1 hour incubation time, ASA caused a decrease in sperm viability, motility, plasma membrane integrity, and inhibit sperm tyrosine phosphorylation. ASA caused a decrease in viability, motility, sperm plasma membrane integrity, and tyrosine phosphorylation of sperm after 1 hour incubation time.Conclusion: ASA from infertile women reduced the sperm viability, motility, plasma membrane integrity, and capacitation in dose and time dependent manner.

scholarly journals Cytotoxic and Apoptotic Activity of Majoranolide from Mezilaurus crassiramea on HL-60 Leukemia Cells

2019 ◽  
Vol 2019 ◽  
pp. 1-8
Author(s):  
Lanna M. Heemann ◽  
Kamylla F. S. de Souza ◽  
Danilo Tófoli ◽  
Kelly J. Filippin ◽  
Walmir S. Garcez ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Plasma Membrane ◽  
Caspase 3 ◽  
Membrane Integrity ◽  
Ethanol Extract ◽  
2D Nmr ◽  
Dependent Manner ◽  
Plasma Membrane Integrity ◽  
Apoptotic Activity ◽  
Apoptotic Pathways

Majoranolide, a butanolide isolated from the nonpolar fraction of an ethanol extract of Mezilaurus crassiramea (Lauraceae) fruits, is being reported for the first time in this genus and the third time in plants. Structurally identified from 1D and 2D NMR and HRESIMS data, majoranolide proved cytotoxic against cancer cells—MCF-7 and MDA-MB-231 (breast), HT-29 (colon), PC-3 (prostate), 786-0 (renal), and HL-60 (leukemia)—inhibiting growth in HL-60 cells (GI50 = 0.21 μM) and exhibiting higher selectivity for this line than for nonneoplastic NIH/3T3 murine fibroblasts. Effects on the cell cycle, caspase-3 activation, and plasma membrane integrity were evaluated by flow cytometry. Expression of genes related to apoptotic pathways (BAX, BCL2, BIRC5, and CASP8) was investigated using RT-qPCR. At 50 μM, majoranolide induced cell cycle arrest at G1 in 24 h increased the sub-G1 population in 48 h and increased caspase-3 activation in a time-dependent manner. The compound upregulated BAX and CASP8 transcription (proapoptotic genes) and downregulated BIRC5 (antiapoptotic). Loss of plasma membrane integrity in 30% of cells occurred at 48 h, but not at 24 h, characterizing gradual, programmed death. The results suggest that majoranolide cytotoxicity involves apoptosis induction in HL-60 cells, although other mechanisms may contribute to this cell death.

scholarly journals Accumulation of Non-Superoxide Anion Reactive Oxygen Species Mediates Nitrogen-Limited Alcoholic Fermentation by Saccharomyces cerevisiae

2010 ◽  
Vol 76 (24) ◽  
pp. 7918-7924 ◽  
Author(s):  
Ana Mendes-Ferreira ◽  
Belém Sampaio-Marques ◽  
Catarina Barbosa ◽  
Fernando Rodrigues ◽  
Vítor Costa ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Cell Cycle ◽  
Plasma Membrane ◽  
Superoxide Anion ◽  
Nitrogen Limitation ◽  
Alcoholic Fermentation ◽  
Membrane Integrity ◽  
Oxygen Species ◽  
Plasma Membrane Integrity ◽  
Cycle Arrest

ABSTRACT Throughout alcoholic fermentation, nitrogen depletion is one of the most important environmental stresses that can negatively affect the yeast metabolic activity and ultimately leads to fermentation arrest. Thus, the identification of the underlying effects and biomarkers of nitrogen limitation is valuable for controlling, and therefore optimizing, alcoholic fermentation. In this study, reactive oxygen species (ROS), plasma membrane integrity, and cell cycle were evaluated in a wine strain of Saccharomyces cerevisiae during alcoholic fermentation in nitrogen-limiting medium under anaerobic conditions. The results indicated that nitrogen limitation leads to an increase in ROS and that the superoxide anion is a minor component of the ROS, but there is increased activity of both Sod2p and Cta1p. Associated with these effects was a decrease in plasma membrane integrity and a persistent cell cycle arrest at G0/G1 phases. Moreover, under these conditions it appears that autophagy, evaluated by ATG8 expression, is induced, suggesting that this mechanism is essential for cell survival but does not prevent the cell cycle arrest observed in slow fermentation. Conversely, nitrogen refeeding allowed cells to reenter cell cycle by decreasing ROS generation and autophagy. Altogether, the results provide new insights on the understanding of wine fermentations under nitrogen-limiting conditions and further indicate that ROS accumulation, evaluated by the MitoTracker Red dye CM-H2XRos, and plasma membrane integrity could be useful as predictive markers of fermentation problems.

scholarly journals ABT-737 Triggers Caspase-Dependent Inhibition of Platelet Procoagulant Extracellular Vesicle Release during Apoptosis and Secondary Necrosis In Vitro

2019 ◽  
Vol 119 (10) ◽  
pp. 1665-1674 ◽  
Author(s):  
Hao Wei ◽  
Matthew T. Harper
Keyword(s):  
Plasma Membrane ◽  
Membrane Integrity ◽  
Annexin V ◽  
Extracellular Vesicle ◽  
Ionophore A23187 ◽  
Dependent Manner ◽  
Plasma Membrane Integrity ◽  
Secondary Necrosis

AbstractPlatelet lifespan is limited by activation of intrinsic apoptosis. Apoptotic platelets are rapidly cleared from the circulation in vivo. ABT-737 triggers platelet apoptosis and is a useful tool for studying this process. However, in vitro experiments lack clearance mechanisms for apoptotic platelets. To determine whether apoptotic platelets progress to secondary necrosis, apoptosis was triggered in human platelets with ABT-737, a BH3 mimetic. Platelet annexin V (AnV) binding, release of AnV+ extracellular vesicles (EVs), and loss of plasma membrane integrity were monitored by flow cytometry. ABT-737 triggered AnV binding, indicating phosphatidylserine exposure, release of AnV+ EVs, and a slow loss of plasma membrane integrity. The latter suggests that apoptotic platelets progress to secondary necrosis in vitro. These responses were dependent on caspase activation and Ca2+ entry. Surprisingly, although intracellular Ca2+ concentration increased, AnV+ EV release was not dependent on the Ca2+-dependent protease, calpain. On the contrary, ABT-737 downregulated the ability of the Ca2+ ionophore, A23187, to trigger calpain-dependent release of AnV+ EVs. This was dependent on caspase activity as, when caspases were inhibited, ABT-737 increased the ability of A23187 to trigger AnV+ EV release. These data suggest that apoptotic platelets progress to secondary necrosis unless they are cleared. This may affect the interpretation of ABT-737-triggered signaling in platelets in vitro. Ca2+-dependent AnV+ EV release is downregulated during apoptosis in a caspase-dependent manner, which may limit the potential consequences of secondary necrotic platelets.

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