Pulsed-laser creation and characterization of giant plasma membrane vesicles from cells

Kelly, Christopher V.; Kober, Mary-Margaret T.; Kinnunen, Päivö; Reis, David A.; Orr, Bradford G.; Holl, Mark M. Banaszak

doi:10.1007/s10867-009-9167-7

Cited by 16 publications

(14 citation statements)

References 45 publications

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“…Large plasma membrane vesicles, or blebs, are clearly noticeable in Figure 3a, and to a lesser extent in Figure 3c and e, indicating the rupture of the cellular plasma membrane–cytoskeleton interface due to the combined effect of the pulse irradiation and the nanoparticles. Similar vesicles were previously observed in cells as a result of tightly focused femtosecond pulse beam irradiation 13, 25…”

Section: Resultssupporting

confidence: 80%

Optical Nanomanipulations of Malignant Cells: Controlled Cell Damage and Fusion

Minai

Yeheskely‐Hayon

Golan

et al. 2012

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Specifically targeting and manipulating living cells is a key challenge in biomedicine and in cancer research in particular. Several studies have shown that nanoparticles irradiated by intense lasers are capable of conveying damage to nearby cells for various therapeutic and biological applications. In this work ultrashort laser pulses and gold nanospheres are used for the generation of localized, nanometric disruptions on the membranes of specifically targeted cells. The high structural stability of the nanospheres and the resonance pulse irradiation allow effective means for controlling the induced nanometric effects. The technique is demonstrated by inducing desired death mechanisms in epidermoid carcinoma and Burkitt lymphoma cells, and initiating efficient cell fusion between various cell types. Main advantages of the presented approach include low toxicity, high specificity, and high flexibility in the regulation of cell damage and cell fusion, which would allow it to play an important role in various future clinical and scientific applications.

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Section: Resultssupporting

confidence: 80%

Optical Nanomanipulations of Malignant Cells: Controlled Cell Damage and Fusion

Minai

Yeheskely‐Hayon

Golan

et al. 2012

Small

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“…Indeed, interfering with the activities of linking molecules such as ezrin / radixin / moesin (ERM) and class I myosins frequently induced blebbing in zebrafish prechordal plate mesendoderm progenitor cells [ 35 ]. Local pulsed-laser-induced ablation of the cell membrane can induce blebbing [ 36 , 37 ], which also supports the above idea. In the present study, virtually no blebs were observed in the anucleate fragments of myosin II-null cells; instead, very small extensions were often observed.…”

Section: Discussionsupporting

confidence: 56%

Microtubule-Mediated Inositol Lipid Signaling Plays Critical Roles in Regulation of Blebbing

2015

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Cells migrate by extending pseudopods such as lamellipodia and blebs. Although the signals leading to lamellipodia extension have been extensively investigated, those for bleb extension remain unclear. Here, we investigated signals for blebbing in Dictyostelium cells using a newly developed assay to induce blebbing. When cells were cut into two pieces with a microneedle, the anucleate fragments vigorously extended blebs. This assay enabled us to induce blebbing reproducibly, and analyses of knockout mutants and specific inhibitors identified candidate molecules that regulate blebbing. Blebs were also induced in anucleate fragments of leukocytes, indicating that this assay is generally applicable to animal cells. After cutting, microtubules in the anucleate fragments promptly depolymerized, followed by the extension of blebs. Furthermore, when intact cells were treated with a microtubule inhibitor, they frequently extended blebs. The depolymerization of microtubules induced the delocalization of inositol lipid phosphatidylinositol 3,4,5-trisphosphate from the cell membrane. PI3 kinase-null cells frequently extended blebs, whereas PTEN-null cells extended fewer blebs. From these observations, we propose a model in which microtubules play a critical role in bleb regulation via inositol lipid metabolism.

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“…[57]) or in some fully-differentiated cells (e.g., inner hair cells [58], skeletal muscle fibers [33]). Once formed, these balloon blebs are typically not retrieved.…”

Section: Discussionmentioning

confidence: 99%

Force Spectroscopy Measurements Show That Cortical Neurons Exposed to Excitotoxic Agonists Stiffen before Showing Evidence of Bleb Damage

et al. 2013

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In ischemic and traumatic brain injury, hyperactivated glutamate (N-methyl-D-aspartic acid, NMDA) and sodium (Nav) channels trigger excitotoxic neuron death. Na+, Ca++ and H2O influx into affected neurons elicits swelling (increased cell volume) and pathological blebbing (disassociation of the plasma membrane’s bilayer from its spectrin-actomyosin matrix). Though usually conflated in injured tissue, cell swelling and blebbing are distinct processes. Around an injury core, salvageable neurons could be mildly swollen without yet having suffered the bleb-type membrane damage that, by rendering channels leaky and pumps dysfunctional, exacerbates the excitotoxic positive feedback spiral. Recognizing when neuronal inflation signifies non-lethal osmotic swelling versus blebbing should further efforts to salvage injury-penumbra neurons. To assess whether the mechanical properties of osmotically-swollen versus excitotoxically-blebbing neurons might be cytomechanically distinguishable, we measured cortical neuron elasticity (gauged via atomic force microscopy (AFM)-based force spectroscopy) upon brief exposure to hypotonicity or to excitotoxic agonists (glutamate and Nav channel activators, NMDA and veratridine). Though unperturbed by solution exchange per se, elasticity increased abruptly with hypotonicity, with NMDA and with veratridine. Neurons then invariably softened towards or below the pre-treatment level, sometimes starting before the washout. The initial channel-mediated stiffening bespeaks an abrupt elevation of hydrostatic pressure linked to NMDA or Nav channel-mediated ion/H2O fluxes, together with increased [Ca++]int-mediated submembrane actomyosin contractility. The subsequent softening to below-control levels is consistent with the onset of a lethal level of bleb damage. These findings indicate that dissection/identification of molecular events during the excitotoxic transition from stiff/swollen to soft/blebbing is warranted and should be feasible.

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Pulsed-laser creation and characterization of giant plasma membrane vesicles from cells

Cited by 16 publications

References 45 publications

Optical Nanomanipulations of Malignant Cells: Controlled Cell Damage and Fusion

Optical Nanomanipulations of Malignant Cells: Controlled Cell Damage and Fusion

Microtubule-Mediated Inositol Lipid Signaling Plays Critical Roles in Regulation of Blebbing

Force Spectroscopy Measurements Show That Cortical Neurons Exposed to Excitotoxic Agonists Stiffen before Showing Evidence of Bleb Damage

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