An Improved Transwell Design for Microelectrode Ion-Flux Measurements

Buchroithner, Boris; Spurny, Pavel; Mayr, Sandra; Heitz, J.; Sivun, Dmitry; Jacak, Jaroslaw; Ludwig, Jost

doi:10.3390/mi12030273

Cited by 4 publications

(2 citation statements)

References 27 publications

(28 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Student's t-tests were conducted to evaluate the results and ascertain the significant difference between means at the significant level of *P<0.05 and represented as mean ± standard error (SE). The MIFE technique has offered a significant conceptual development as well as advancement to understand the responses and cellular membrane transport processes of plants, fungi, bacteria, animal cells, and biofilms related to different environmental stress conditions (Shabala et al, 2012;Buchroithner et al, 2021). Newman (2001), an Australian scientist, invented this technique in 2001 by using ion-selective microelectrodes to determine the non-invasive measurement of ion fluxes (Figure S5).…”

Section: Discussionmentioning

confidence: 99%

AtCIPK16, a CBL-interacting protein kinase gene, confers salinity tolerance in transgenic wheat

et al. 2023

View full text Add to dashboard Cite

Globally, wheat is the major source of staple food, protein, and basic calories for most of the human population. Strategies must be adopted for sustainable wheat crop production to fill the ever-increasing food demand. Salinity is one of the major abiotic stresses involved in plant growth retardation and grain yield reduction. In plants, calcineurin-B-like proteins form a complicated network with the target kinase CBL-interacting protein kinases (CIPKs) in response to intracellular calcium signaling as a consequence of abiotic stresses. The AtCIPK16 gene has been identified in Arabidopsis thaliana and found to be significantly upregulated under salinity stress. In this study, the AtCIPK16 gene was cloned in two different plant expression vectors, i.e., pTOOL37 having a UBI1 promoter and pMDC32 having a 2XCaMV35S constitutive promoter transformed through the Agrobacterium-mediated transformation protocol, in the local wheat cultivar Faisalabad-2008. Based on their ability to tolerate different levels of salt stress (0, 50, 100, and 200 mM), the transgenic wheat lines OE1, OE2, and OE3 expressing AtCIPK16 under the UBI1 promoter and OE5, OE6, and OE7 expressing the same gene under the 2XCaMV35S promoter performed better at 100 mM of salinity stress as compared with the wild type. The AtCIPK16 overexpressing transgenic wheat lines were further investigated for their K+ retention ability in root tissues by utilizing the microelectrode ion flux estimation technique. It has been demonstrated that after 10 min of 100 mM NaCl application, more K+ ions were retained in the AtCIPK16 overexpressing transgenic wheat lines than in the wild type. Moreover, it could be concluded that AtCIPK16 functions as a positive elicitor in sequestering Na+ ions into the cell vacuole and retaining more cellular K+ under salt stress to maintain ionic homeostasis.

show abstract

Section: Discussionmentioning

confidence: 99%

AtCIPK16, a CBL-interacting protein kinase gene, confers salinity tolerance in transgenic wheat

et al. 2023

View full text Add to dashboard Cite

show abstract

“…After placing the petri dish with the sample into MIFE TM 3 system (University of Tasmania), measuring microelectrodes (K + and Na + ) were adjusted just close above the monolayer of the cells by MIFE custom-assembled Narishige manipulator system consisting of MHW-4 micromanipulator (5:1 movement) and SM-17 electrode holder (Narishige, Tokyo, Japan). Movement of electrodes was provided by MIFE-MDRIVE motor drive system (complete) for hydraulic micromanipulator 65 . Samples and electrodes were observed by inverted Zeiss PrimoVert microscope.…”

Section: Methodsmentioning

confidence: 99%

Targeting plasma membrane and mitochondrial instability in breast cancer cells and breast epithelial to mesenchymal transition-model cells by adamantyl diaza-crown ether ZG613

Ester

Mioč

Spurny

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

The adamantane derived diaza-crown ether ZG613 was assessed as a potential breast cancer cells and breast epithelial to mesenchymal transition (EMT)-model cells targeting agent. We postulated that ZG613 activity relies on its plasma/mitochondria membrane disruption ability based on adamantane hydrophobicity and/or crown ether related ionophoric properties. We performed molecular dynamics (MD) simulations and next generation sequencing, followed by in vitro study of cell death, membrane perturbations and ionophoric ability, as well as in vivo study of effects on the tumour growth. MD simulation and RNA sequencing pointed toward physical disruption of plasma membrane by ZG613, corroborated by measured increase in membrane permeability leading to cell death. Measurements of ion fluxes confirmed ZG613 inability to transport Na+ and K+, as predicted by MD simulation. EMT-model cells exhibit changes in mitochondrial morphology and ATP levels, successfully targeted by ZG613. ZG613 caused mild retardation of tumour growth in vivo. In conclusion, ZG613 kills breast cancer cells and breast EMT-model cells by physical disruption of plasma membrane and impairments of mitochondrial functions. Breast EMT cells represent good potential targets within the breast tumour, due to their plasma membrane and mitochondrial instability.

show abstract

Laser Structuring for Biomedical Applications

Buchberger

Muck

Plamadeala

et al. 2023

Springer Series in Optical Sciences

View full text Add to dashboard Cite

Laser structuring enables modification of sample topography, surface chemistry, and/or physical properties of materials. Examples of these processes are ripple, nap or wall formation, surface oxidation, induction of polymerization reactions, or changes in crystallinity or contact angle. These – most of the time – interrelated modifications are exploited widely for biomedical applications. They range from cell-repellent surfaces for easy-to-replace cardiac pacemakers, control of cell proliferation required in regenerative medicine, to increased cell adhesion for cell arrays. Furthermore, ns-laser-induced nanoripples were used for formation of gold nanowires for future surface plasmon resonance sensors directly integrated into biotechnological devices. Additive nano- and microscale manufacturing by two-photon polymerization allows for considerable progress in cell scaffold formation, paving the path for in vitro–grown organs, bones, and cartilages. The very same fs-laser-based technique was also used for biomimetic microneedles with enhanced liquid spreading on their surface. Microneedles are promising candidates for low-cost, high-throughput drug delivery and vaccination applicable even by nonmedically trained personnel. Microfluidic systems fabricated by fs-lasers have enabled progress in 3D microscopy of single cells and in studies on thrombocyte activation with the help of nanoanchors. Explicating the abovementioned and further biomedical applications, the authors put special focus on the achieved limits pointing out what scientists have accomplished so far in their pursuit of extreme scales.

show abstract

An Improved Transwell Design for Microelectrode Ion-Flux Measurements

Cited by 4 publications

References 27 publications

AtCIPK16, a CBL-interacting protein kinase gene, confers salinity tolerance in transgenic wheat

AtCIPK16, a CBL-interacting protein kinase gene, confers salinity tolerance in transgenic wheat

Targeting plasma membrane and mitochondrial instability in breast cancer cells and breast epithelial to mesenchymal transition-model cells by adamantyl diaza-crown ether ZG613

Laser Structuring for Biomedical Applications

Contact Info

Product

Resources

About