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Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation
Published on: February 8, 2015
Correlation between the loss of intracellular molecules and cell viability after cell electroporation
Baltramiejus Jakstys1, Milda Jakutaviciute1, Dovile Uzdavinyte1
1Faculty of Natural Sciences, Vytautas Magnus University, Kaunas LT 44404, Lithuania.
Abstract:
Control of membrane permeability to exogenous compounds by membrane electroporation can lead to cell death, which is related to permanent membrane damage, oxidation stress, leakage of intracellular molecules. In this study, we show that the predominant cell death modality after the application of high voltage electric pulses is related with inability to reseal of initial pores (first stage irreversible electroporation, FirEP). After moderately strong electric pulses, initial pores reseal, however, some cell still die later on due to electric field induced cell stress which leads to delayed cell death (late-stage irreversible electroporation, LirEP). According to our data, the period in which the majority of cells commit to either pore resealing or complete loss of barrier function depends on the intensity of electric field treatment but did not exceed 35 min. Additionally, we show that after electroporation using electric pulse parameters that induce LirEP, some cells can be rescued by supplementing medium with compounds obtained from irreversibly electroporated cells. We determined that the intracellular molecules that contribute to the increase of cell viability are larger than 30 kDa. This serves to prove that the loss of intracellular compounds plays a significant role in the decrease of cell viability after electroporation.
Insights
Electroporation can cause cell death through pore damage. This study identifies two types: immediate pore resealing failure (FirEP) and delayed cell death (LirEP) from stress, with larger molecules rescuing cells.
Area of Science:
- Cell biology
- Biophysics
- Electroporation
Background:
- Membrane electroporation controls permeability but can cause cell death via membrane damage, oxidative stress, and molecule leakage.
- Understanding cell death mechanisms post-electroporation is crucial for optimizing its applications.
Purpose of the Study:
- To investigate the predominant cell death modalities following high and moderate voltage electric pulses.
- To determine the time frame for cell fate commitment after electroporation.
- To explore rescue strategies for cells undergoing delayed electroporation-induced death.
Main Methods:
- Application of high and moderate voltage electric pulses to cells.
- Analysis of cell death mechanisms, including pore resealing and stress responses.
- Investigating cell rescue using supplements from electroporated cells.
Main Results:
- High voltage pulses primarily cause immediate cell death due to irreversible pore damage (FirEP).
- Moderate pulses lead to delayed cell death (LirEP) from electric field-induced stress, with cell fate determined within 35 minutes.
- Supplementing medium with molecules >30 kDa from electroporated cells can rescue cells undergoing LirEP, indicating loss of intracellular compounds is critical.
Conclusions:
- Electroporation-induced cell death is modality-dependent, with FirEP and LirEP representing distinct pathways.
- Cellular recovery from LirEP is possible through the reintroduction of specific intracellular molecules.
- The findings highlight the significant role of intracellular compound loss in electroporation-induced cell death.
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