scholarly journals Imaging the electrical activity of organelles in living cells

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Ella Matamala ◽  
Cristian Castillo ◽  
Juan P. Vivar ◽  
Patricio A. Rojas ◽  
Sebastian E. Brauchi
Keyword(s):  
Electrical Activity ◽  
Living Cells ◽  
Eukaryotic Cells ◽  
Subcellular Targeting ◽  
Spatiotemporal Resolution ◽  
Non Invasive ◽  
Optical Multiplexing ◽  
Voltage Dependent ◽  
Membrane Bound ◽  
Fret Pair

AbstractEukaryotic cells are complex systems compartmentalized in membrane-bound organelles. Visualization of organellar electrical activity in living cells requires both a suitable reporter and non-invasive imaging at high spatiotemporal resolution. Here we present hVoSorg, an optical method to monitor changes in the membrane potential of subcellular membranes. This method takes advantage of a FRET pair consisting of a membrane-bound voltage-insensitive fluorescent donor and a non-fluorescent voltage-dependent acceptor that rapidly moves across the membrane in response to changes in polarity. Compared to the currently available techniques, hVoSorg has advantages including simple and precise subcellular targeting, the ability to record from individual organelles, and the potential for optical multiplexing of organellar activity.

scholarly journals Imaging the Electrical Activity Of Organelles in living cells

2019 ◽  
Author(s):  
Ella Matamala ◽  
Cristian Castillo ◽  
Juan Pablo Vivar ◽  
Sebastian Brauchi
Keyword(s):  
Membrane Potential ◽  
Electrical Activity ◽  
Living Cells ◽  
Resting Membrane Potential ◽  
Subcellular Targeting ◽  
Spatiotemporal Resolution ◽  
Non Invasive ◽  
Optical Multiplexing ◽  
Voltage Dependent ◽  
Membrane Bound

AbstractEukaryotic cells are complex systems compartmentalized in membrane-bound organelles. Visualization of organellar electrical activity in living cells requires both a suitable reporter and non-invasive imaging at high spatiotemporal resolution. Here we present hVoSorg, an optical method to monitor changes in the membrane potential of subcellular membranes. This method takes advantage of a FRET pair consisting of a membrane-bound voltage-insensitive fluorescent donor and a colorless voltage-dependent acceptor that rapidly moves across the membrane in response to changes in polarity. Compared to the currently available techniques, hVoSorg has advantages including simple and precise subcellular targeting, the ability to record from individual organelles, and the potential for optical multiplexing.SummaryBy adapting a hybrid-FRET voltage sensor we report here the resting membrane potential of different organelles in living cells.

scholarly journals An engineered membrane-bound guanylyl cyclase with light-switchable activity

BMC Biology ◽  
2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Yuehui Tian ◽  
Georg Nagel ◽  
Shiqiang Gao
Keyword(s):  
Guanylyl Cyclase ◽  
Green Light ◽  
Chemical Properties ◽  
Light Sensitivity ◽  
Cgmp Level ◽  
Volvox Carteri ◽  
Non Invasive ◽  
Cgmp Signaling ◽  
Membrane Bound ◽  
Light Flashes

Abstract Background Microbial rhodopsins vary in their chemical properties, from light sensitive ion transport to different enzymatic activities. Recently, a novel family of two-component Cyclase (rhod)opsins (2c-Cyclop) from the green algae Chlamydomonas reinhardtii and Volvox carteri was characterized, revealing a light-inhibited guanylyl cyclase (GC) activity. More genes similar to 2c-Cyclop exist in algal genomes, but their molecular and physiological functions remained uncharacterized. Results Chlamyopsin-5 (Cop5) from C. reinhardtii is related to Cr2c-Cyclop1 (Cop6) and can be expressed in Xenopus laevis oocytes, but shows no GC activity. Here, we exchanged parts of Cop5 with the corresponding ones of Cr2c-Cyclop1. When exchanging the opsin part of Cr2c-Cyclop1 with that of Cop5, we obtained a bi-stable guanylyl cyclase (switch-Cyclop1) whose activity can be switched by short light flashes. The GC activity of switch-Cyclop1 is increased for hours by a short 380 nm illumination and switched off (20-fold decreased) by blue or green light. switch-Cyclop1 is very light-sensitive and can half-maximally be activated by ~ 150 photons/nm2 of 380 nm (~ 73 J/m2) or inhibited by ~ 40 photons/nm2 of 473 nm (~ 18 J/m2). Conclusions This engineered guanylyl cyclase is the first light-switchable enzyme for cGMP level regulation. Light-regulated cGMP production with high light-sensitivity is a promising technique for the non-invasive investigation of the effects of cGMP signaling in many different tissues.

scholarly journals Mitochondrial Protein Network: From Biogenesis to Bioenergetics in Health and Disease

2020 ◽  
Vol 22 (1) ◽  
pp. 1
Author(s):  
Alessandra Ferramosca
Keyword(s):  
Crucial Role ◽  
Mitochondrial Protein ◽  
Protein Network ◽  
Eukaryotic Cells ◽  
Double Membrane ◽  
Health And Disease ◽  
Membrane Bound

Mitochondria are double membrane-bound organelles which are essential for the viability of eukaryotic cells, because they play a crucial role in bioenergetics, metabolism and signaling [...]

scholarly journals The “Vesicular Intelligence” Strategy of Blood Cancers

Genes ◽  
2021 ◽  
Vol 12 (3) ◽  
pp. 416
Author(s):  
Dorian Forte ◽  
Martina Barone ◽  
Francesca Palandri ◽  
Lucia Catani
Keyword(s):  
Clonal Evolution ◽  
Biological Properties ◽  
Hemopoietic Stem Cell ◽  
Target Cells ◽  
Cancer Cell Growth ◽  
Cell Compartment ◽  
Blood Cancer ◽  
Non Invasive ◽  
Membrane Bound ◽  
Immune Potential

Blood cancers are a heterogeneous group of disorders including leukemia, multiple myeloma, and lymphoma. They may derive from the clonal evolution of the hemopoietic stem cell compartment or from the transformation of progenitors with immune potential. Extracellular vesicles (EVs) are membrane-bound nanovesicles which are released by cells into body fluids with a role in intercellular communication in physiology and pathology, including cancer. EV cargos are enriched in nucleic acids, proteins, and lipids, and these molecules can be delivered to target cells to influence their biological properties and modify surrounding or distant targets. In this review, we will describe the “smart strategy” on how blood cancer-derived EVs modulate tumor cell development and maintenance. Moreover, we will also depict the function of microenvironment-derived EVs in blood cancers and discuss how the interplay between tumor and microenvironment affects blood cancer cell growth and spreading, immune response, angiogenesis, thrombogenicity, and drug resistance. The potential of EVs as non-invasive biomarkers will be also discussed. Lastly, we discuss the clinical application viewpoint of EVs in blood cancers. Overall, blood cancers apply a ‘vesicular intelligence’ strategy to spread signals over their microenvironment, promoting the development and/or maintenance of the malignant clone.

A novel microfluidic microelectrode chip for a significantly enhanced monitoring of NPY-receptor activation in live mode

Lab on a Chip ◽  
2017 ◽  
Vol 17 (24) ◽  
pp. 4294-4302 ◽  
Author(s):  
Franziska D. Zitzmann ◽  
Heinz-Georg Jahnke ◽  
Felix Nitschke ◽  
Annette G. Beck-Sickinger ◽  
Bernd Abel ◽  
...  
Keyword(s):  
Real Time ◽  
Microfluidic Chip ◽  
Microelectrode Array ◽  
Fem Simulation ◽  
Receptor Activation ◽  
Living Cells ◽  
Real Time Monitoring ◽  
Non Invasive ◽  
Npy Receptor ◽  
Simulation Based

We present a FEM simulation based step-by-step development of a microelectrode array integrated into a microfluidic chip for the non-invasive real-time monitoring of living cells.

Biomems Microprobe for Electrical Activity Recording of Living Cells

2007 ◽  
Author(s):  
C. Moldovan ◽  
R. Iosub ◽  
C.P. Lungu ◽  
A.M. Lungu ◽  
B. Firtat ◽  
...  
Keyword(s):  
Electrical Activity ◽  
Living Cells

Optobiochemistry: Genetically Encoded Control of Protein Activity by Light

2021 ◽  
Vol 90 (1) ◽  
Author(s):  
Jihye Seong ◽  
Michael Z. Lin
Keyword(s):  
Protein Function ◽  
Living Cells ◽  
Optical Methods ◽  
Annual Review ◽  
Publication Date ◽  
Protein Activity ◽  
Spatiotemporal Resolution ◽  
Protein Functions ◽  
Protein Classes ◽  
Control Protein

Optobiochemical control of protein activities allows the investigation of protein functions in living cells with high spatiotemporal resolution. Over the last two decades, numerous natural photosensory domains have been characterized and synthetic domains engineered and assembled into photoregulatory systems to control protein function with light.Here, we review the field of optobiochemistry, categorizing photosensory domains by chromophore, describing photoregulatory systems by mechanism of action, and discussing protein classes frequently investigated using optical methods. We also present examples of how spatial or temporal control of proteins in living cells has provided new insights not possible with traditional biochemical or cell biological techniques. Expected final online publication date for the Annual Review of Biochemistry, Volume 90 is June 2021. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

scholarly journals Dynamic nanoscale morphology of the ER surveyed by STED microscopy

2018 ◽  
Vol 218 (1) ◽  
pp. 83-96 ◽  
Author(s):  
Lena K. Schroeder ◽  
Andrew E.S. Barentine ◽  
Holly Merta ◽  
Sarah Schweighofer ◽  
Yongdeng Zhang ◽  
...  
Keyword(s):  
Living Cells ◽  
Stimulated Emission ◽  
Membrane Structures ◽  
Spatiotemporal Resolution ◽  
Sted Microscopy ◽  
Superresolution Microscopy ◽  
Structure And Dynamics ◽  
First Time ◽  
Nanoscale Dynamics ◽  
Conventional Light Microscopy

The endoplasmic reticulum (ER) is composed of interconnected membrane sheets and tubules. Superresolution microscopy recently revealed densely packed, rapidly moving ER tubules mistaken for sheets by conventional light microscopy, highlighting the importance of revisiting classical views of ER structure with high spatiotemporal resolution in living cells. In this study, we use live-cell stimulated emission depletion (STED) microscopy to survey the architecture of the ER at 50-nm resolution. We determine the nanoscale dimensions of ER tubules and sheets for the first time in living cells. We demonstrate that ER sheets contain highly dynamic, subdiffraction-sized holes, which we call nanoholes, that coexist with uniform sheet regions. Reticulon family members localize to curved edges of holes within sheets and are required for their formation. The luminal tether Climp63 and microtubule cytoskeleton modulate their nanoscale dynamics and organization. Thus, by providing the first quantitative analysis of ER membrane structure and dynamics at the nanoscale, our work reveals that the ER in living cells is not limited to uniform sheets and tubules; instead, we suggest the ER contains a continuum of membrane structures that includes dynamic nanoholes in sheets as well as clustered tubules.

scholarly journals The N-Terminal Segment of the Voltage-Dependent Anion Channel: A Possible Membrane-Bound Intermediate in Pore Unbinding

2019 ◽  
Vol 431 (2) ◽  
pp. 223-243 ◽  
Author(s):  
Maria M. Reif ◽  
Moritz Fischer ◽  
Kai Fredriksson ◽  
Franz Hagn ◽  
Martin Zacharias
Keyword(s):  
Anion Channel ◽  
Terminal Segment ◽  
Voltage Dependent Anion Channel ◽  
Voltage Dependent ◽  
Membrane Bound

scholarly journals Super-Resolution Imaging by Dual Iterative Structured Illumination Microscopy

2021 ◽  
Author(s):  
Anna Loeschberger ◽  
Yauheni Novikau ◽  
Ralf Netz ◽  
Marie-Christin Spindler ◽  
Ricardo Benavente ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Brain Slices ◽  
Super Resolution ◽  
Reconstruction Algorithm ◽  
Living Cells ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Spatiotemporal Resolution ◽  
Coated Pits ◽  
Resolution Imaging

Three-dimensional (3D) multicolor super-resolution imaging in the 50-100 nm range in fixed and living cells remains challenging. We extend the resolution of structured illumination microscopy (SIM) by an improved nonlinear iterative reconstruction algorithm that enables 3D multicolor imaging with improved spatiotemporal resolution at low illumination intensities. We demonstrate the performance of dual iterative SIM (diSIM) imaging cellular structures in fixed cells including synaptonemal complexes, clathrin coated pits and the actin cytoskeleton with lateral resolutions of 60-100 nm with standard fluorophores. Furthermore, we visualize dendritic spines in 70 micrometer thick brain slices with an axial resolution < 200 nm. Finally, we image dynamics of the endoplasmatic reticulum and microtubules in living cells with up to 255 frames/s.

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