Tag Archives: uniqprobes

Cytoskeletal disorganization underlies PABPN1-mediated myogenic disability

Muscle wasting is connected with changes in various cellular mechanisms that influence protein homeostasis, transcription, protein acetylation and different metabolic pathways. *

Scientific studies have shown that reduced levels of polyadenylation binding protein 1 ( PABPN1 , a multifactorial regulator of mRNA processing ) cause muscle wasting, including muscle atrophy, extracellular matrix thickening, myofiber typing transitions and central nucleation. *

However, the cellular mechanisms behind PABPN1-mediated muscle wasting are not fully understood. *

In the article “Cytoskeletal disorganization underlies PABPN1-mediated myogenic disability” Cyriel Sebastiaan Olie, Erik van der Wal, Cikes Domagoj, Loes Maton, Jessica C. de Greef, I.-Hsuan Lin, Yi-Fan Chen, Elsayad Kareem, Josef M. Penninger, Benedikt M. Kessler and Vered Raz examine the cytoskeletal auxiliary changes that are dependent on PABPN1 levels using 2D and 3D models, and investigate how these affect muscle wasting. *

They suggest that poor cytoskeletal mechanical features are caused by altered expression levels of cytoskeletal proteins and contribute to muscle wasting and atrophy. *

For the measurements of cell-mechanics properties in control and shPAB cells ( muscle cells with reduced PABPN1 levels ) the authors used Brillouin Light Scattering Microscopy and Atomic Force Microscopy. *

NANOSENSORS™ uniqprobe qp-BioAC ( CB3 ) AFM probes were used in the quantitative imaging where a force curve is applied at each point. The analyzed area of each cell was 5 µm × 5 µm (64 × 64 pixels) with an approach speed of 35 µm/s (3.4 ms/pixel), and applied forces of up to 118 pN. *

Figure 4 from “Cytoskeletal disorganization underlies PABPN1-mediated myogenic disability” by Cyriel Sebastiaan Olie et al.
 Disrupted cytoskeletal spatial organization in shPAB human muscle cell cultures. A Representative images of control and shPAB human muscle cell cultures, stained with antibodies to tubulin and actin, and the actin filaments were visualized with actin-GFP. B Tubulin staining in control and shPAB myoblast cell cultures after DMSO, 100 nM nocodazole or 25 nM paclitaxel treatment for 2 h. Scale bar is 25 µm. C Measurements of cell-mechanics properties in control and shPAB cells using the Brillouin Light Scattering Microscopy (Ci) or the Atomic Force Microscopy (Cii). Measurements were carried out in myoblasts; every dot represents the median from 1000 measurements in a cell. Cell stiffness is measured by GHz, and the young modulus reports the cell surface tension. Averages and standard deviations are from N = 15 cells. Statistical significance was calculated with the student’s t-test.
NANOSENSORS uniqprobe qp-BioAC ( CB3 ) AFM probes were used in the quantitative imaging of cell-mechanics properties.
Figure 4 from “Cytoskeletal disorganization underlies PABPN1-mediated myogenic disability” by Cyriel Sebastiaan Olie et al.
 Disrupted cytoskeletal spatial organization in shPAB human muscle cell cultures. A Representative images of control and shPAB human muscle cell cultures, stained with antibodies to tubulin and actin, and the actin filaments were visualized with actin-GFP. B Tubulin staining in control and shPAB myoblast cell cultures after DMSO, 100 nM nocodazole or 25 nM paclitaxel treatment for 2 h. Scale bar is 25 µm. C Measurements of cell-mechanics properties in control and shPAB cells using the Brillouin Light Scattering Microscopy (Ci) or the Atomic Force Microscopy (Cii). Measurements were carried out in myoblasts; every dot represents the median from 1000 measurements in a cell. Cell stiffness is measured by GHz, and the young modulus reports the cell surface tension. Averages and standard deviations are from N = 15 cells. Statistical significance was calculated with the student’s t-test.

*Cyriel Sebastiaan Olie, Erik van der Wal, Cikes Domagoj, Loes Maton, Jessica C. de Greef, I.-Hsuan Lin, Yi-Fan Chen, Elsayad Kareem, Josef M. Penninger, Benedikt M. Kessler & Vered Raz
Cytoskeletal disorganization underlies PABPN1-mediated myogenic disability
Nature Scientific Reports volume 10, Article number: 17621 (2020)
DOI: https://doi.org/10.1038/s41598-020-74676-8

Please follow this external link to read the full article https://rdcu.be/ceyJ4

Open Access: The article “Cytoskeletal disorganization underlies PABPN1-mediated myogenic disability” by Cyriel Sebastiaan Olie, Erik van der Wal, Cikes Domagoj, Loes Maton, Jessica C. de Greef, I.-Hsuan Lin, Yi-Fan Chen, Elsayad Kareem, Josef M. Penninger, Benedikt M. Kessler & Vered Raz is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

Yeast Nanometric Scale Oscillations Highlights Fibronectin Induced Changes in C. Albicans

Yeast resistance to antifungal drugs is a major public health issue. Fungal adhesion onto the host mucosal surface is still a partially unknown phenomenon that is modulated by several actors among which fibronectin plays an important role. Targeting the yeast adhesion onto the mucosal surface could lead to potentially highly efficient treatments. *

In the article “Yeast Nanometric Scale Oscillations Highlights Fibronectin Induced Changes in C. Albicans” Anne-Céline Kohler, Leonardo Venturelli, Abhilash Kannan, Dominique Sanglard, Giovanni Dietler, Ronnie Willaert and Sandor Kasas, explore the effect of fibronectin on the nanomotion pattern of different Candida albicans strains by atomic force microscopy ( AFM )-based nanomotion detection and correlated the cellular oscillations to the yeast adhesion onto epithelial cells. *

The authors demonstrate the ability of nanomotion detection to monitor in real time and in a label-free manner cellular activity changes induced by interacting ligands. Activity changes induced by increasing glucose concentration were observed for Escherichia coli in a previous study. This technique opens novel avenues to detect cellular activation or inhibition induced by ligand–receptor interactions. *

The AFM cantilever oscillations were collected in real time using an by the authors in-house developed nanomotion detection device with NANOSENSORS™ tipless uniqprobe SD-qp-CONT-TL AFM probes from the NANOSENSORS Special Developments List. *

The SD-qp-CONT-TL AFM cantilevers with a nominal spring constant of 0.1 N/m and an average resonant peak in liquids of 8 kHz, were coated with 2 mg/mL of concanavalin A (Con A) for 30 min at room temperature. After removing the excess of Con A, the yeast cells were placed in contact with the AFM cantilever for 1 h at room temperature to allow them to attach to its surface. Poorly attached C. albicans cells were removed by washing gently with YPD medium. Finally, the C. albicans covered AFM cantilever was inserted into the analysis chamber containing 2 mL of filtered (0.2 µm syringe filter, YPD medium. The measurements were performed at room temperature in YPD medium and in YPD medium containing 25 µg/mL of fibronectin. Fibronectin was directly added inside the chip reservoir. For the experiments performed with antifungals, caspofungin was diluted in the YPD present in the analysis chamber to reach a final concentration of 100 µg/mL. *

Supplementary Figure S2 from “Yeast Nanometric Scale Oscillations Highlights Fibronectin Induced Changes in C. Albicans” by Anne-Céline Kohler et al.:
Density of yeast cells on cantilever at the start of the experiment (A) and at the end of the experiment (B). The size of the AFM cantilever is 40 by 130 μm. NANOSENSORS tipless uniqprobe SD-qp-CONT-TL AFM probes from the NANOSENSORS Special Developments List were used in the in-house developed nanomotion detection device.
Supplementary Figure S2 from “Yeast Nanometric Scale Oscillations Highlights Fibronectin Induced Changes in C. Albicans” by Anne-Céline Kohler et al.:
Density of yeast cells on cantilever at the start of the experiment (A) and at the end of the experiment (B). The size of the AFM cantilever is 40 by 130 μm.

*Anne-Céline Kohler, Leonardo Venturelli, Abhilash Kannan, Dominique Sanglard, Giovanni Dietler, Ronnie Willaert and Sandor Kasas
Yeast Nanometric Scale Oscillations Highlights Fibronectin Induced Changes in C. Albicans
Fermentation 2020, 6(1), 28
DOI: https://doi.org/10.3390/fermentation6010028

Please follow this external link to read the full article: https://www.mdpi.com/2311-5637/6/1/28/htm#B5-fermentation-06-00028

Open Access The article “Yeast Nanometric Scale Oscillations Highlights Fibronectin Induced Changes in C. Albicans” by Anne-Céline Kohler, Leonardo Venturelli, Abhilash Kannan, Dominique Sanglard, Giovanni Dietler, Ronnie Willaert and Sandor Kasas is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

Plant-Based Scaffolds Modify Cellular Response to Drug and Radiation Exposure Compared to Standard Cell Culture Models

Plant-based scaffolds present many advantages over a variety of biomaterials.*

Recent studies explored their potential to be repopulated with human cells and thus highlight a growing interest for their use in tissue engineering or for biomedical applications. However, it is still unclear if these in vitro plant-based scaffolds can modify cell phenotype or affect cellular response to external stimuli.

In the research article “Plant-Based Scaffolds Modify Cellular Response to Drug and Radiation Exposure Compared to Standard Cell Culture Models “ Jerome Lacombe, Ashlee F. Harris, Ryan Zenhausern, Sophia Karsunsky and Frederic Zenhausern report the characterization of the mechano-regulation of melanoma SK-MEL-28 and prostate PC3 cells seeded on decellularized spinach leaves scaffolds, compared to cells deposited on standard rigid cell culture substrate, as well as their response to drug and radiation treatment.*

In their study the authors show that plant decellularization provide soft scaffolds that match the stiffness range of most of the human tissue and modify cell behavior, including drug and radiation response, compared to standard cell culture models. Because of their distinguished features (natural vasculature, low immunogenicity, low cost, relative ease, etc.) and their wide variations in the shape and structures, these scaffolds offer a multi-controllable model with multiple biochemical and biophysical interactions. However, additional studies are required to determine if they could address important architectural and physical challenges of the in vivo tissue environment.

For force measurement, the Young’s Modulus (YM) of the leaf scaffolds were determined using force spectroscopy mode at liquid interface with NANOSENSORS uniqprobe qp-BioAC AFM probes for leaves measurement.*

NANOSENSORS uniqprobe qp-BioAC AFM probe top view (SEM image
NANOSENSORS uniqprobe qp-BioAC AFM probe top view (SEM image)

*Jerome Lacombe, Ashlee F. Harris, Ryan Zenhausern, Sophia Karsunsky and Frederic Zenhausern
Plant-Based Scaffolds Modify Cellular Response to Drug and Radiation Exposure Compared to Standard Cell Culture Models
Frontiers in Bioengineering and Biotechnology (2020) 8:932.
DOI: 10.3389/fbioe.2020.00932

Please follow this external link to read the full article: https://www.frontiersin.org/articles/10.3389/fbioe.2020.00932/full#B27

Open Access: The article “Plant-Based Scaffolds Modify Cellular Response to Drug and Radiation Exposure Compared to Standard Cell Culture Models” by Jerome Lacombe, Ashlee F. Harris, Ryan Zenhausern, Sophia Karsunsky and Frederic Zenhausern is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.