CLARITY analysis of the Cl/pH sensor expression in the brain of transgenic mice

Diuba, Artem V.; Samigullin, Dmitry; Kaszás, Attila; Zonfrillo, Francesca; Malkov, Anton; Petukhova, Elena; Casini, Antonio; Arosio, Daniele; Esclapez, Monique; Gross, Cornelius; Bregestovski, Pìotr

doi:10.1016/j.neuroscience.2019.07.010

Cited by 7 publications

(7 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Experiments were performed on laboratory ICR CD-1 outbred mice of both genders and transgenic mice, strain C57BL/6N, expressing ClopHensor [13]. All animal protocols and experimental procedures were approved by the Local Ethics Committee of Kazan State Medical University (N742.…”

Section: Animalsmentioning

confidence: 99%

“…In experiments aimed to monitor concentrations of intracellular Cl − and H + in neurons of brain slices, we used transgenic mice expressing ClopHensor [13]. This biosensor consists of a modified enhanced green fluorescent protein E 2 GFP (ion sensitive) linked to monomeric DsRed (mDsRed) through a flexible 20 amino acid linker [14][15][16].…”

Section: Three Wavelength Excitation Recording Of CL − and H + In Neu...mentioning

confidence: 99%

“…The program allows to selects device operating modes that provide reliable changes in illumination wavelength and intensity, camera image capture, and online graphical visualization of fluorescent signal amplitudes or ratiometric data. Using brain slices from wild-type and transgenic mice expressing a genetically encoded biosensor of hydrogen and chloride [13], we successfully tested the performance of the software using two experimental models: i) simultaneous monitoring of intracellular chloride (Cl − ) and hydrogen (H + ) during synaptic stimulation; ii) recording changes in reactive oxygen species during synaptic stimulation of mouse hippocampal neurons.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

DriveLEDs: software for synchronous control and video acquisition of fluorescent signals

Zakharov,

Ponomareva,

Petukhova

et al. 2023

Front. Phys.

Self Cite

View full text Add to dashboard Cite

Current research uses various fluorescent biosensors to measure ion concentrations, neuronal activity, or cellular components in biological preparations. Several free solutions are available to researchers for recording or analysing fluorescent signals. However, when using different software packages, there are great difficulties in converting data between them. Problems also arise with linking and coordination of individual hardware devices into a single measurement system. Our paper presents useful software that allows to avoid most of these problems. It enables the recording, online visualization and preliminary analysis of fluorescent signals in brain cells and other experimental models. We describe and test software optimized for ratiometric measurements. The program selects device operating modes that allow reliable changes in illumination wavelength, camera image capture, and online graphical visualization of fluorescent signal amplitudes or ratiometric data. The performance of the software was successfully tested on mouse brain using two experimental models; i) simultaneous monitoring of intracellular chloride and hydrogen in transgenic mice expressing genetically encoded biosensor; ii) recording changes in reactive oxygen species during synaptic stimulation of neurons in mouse hippocampal slices. This software allows to overcome the incompatibility of the devices used and reduce the cost of experimental measurements. The software is completely original, easy to use and may be of interest to many scientists involved in the analysis of light-controlled signals in a variety of experimental models, including drug screening, epilepsy models, and other applications. The software is open-source product and can be obtained via GitHub: https://github.com/AndreyZakharovExp/DriveLEDs.

show abstract

Section: Animalsmentioning

confidence: 99%

Section: Three Wavelength Excitation Recording Of CL − and H + In Neu...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

DriveLEDs: software for synchronous control and video acquisition of fluorescent signals

Zakharov,

Ponomareva,

Petukhova

et al. 2023

Front. Phys.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Ventricle post-mortem size assessment was done with methylene blue staining according to methods previously described [23]. Clarity analysis was done according to standard protocol [24].…”

Section: Immunohistochemistrymentioning

confidence: 99%

Investigation and modulation of interleukin-6 following subarachnoid hemorrhage: targeting inflammatory activation for cerebral vasospasm

Lucke‐Wold

Dodd

Motwani

et al. 2022

J Neuroinflammation

View full text Add to dashboard Cite

Background: Cerebral vasospasm (CV) can contribute to significant morbidity in subarachnoid hemorrhage (SAH) patients. A key unknown is how CV induction is triggered following SAH.Methods: Human aneurysmal blood and cerebral spinal fluid were collected for evaluation. To confirm mechanism, c57/bl6 wild type and c57/bl6 IL-6 female knockout (KO) mice were utilized with groups: saline injected, SAH, SAH + IL-6 blockade, SAH IL-6 KO, SAH IL-6 KO + IL-6 administration, SAH + p-STAT3 inhibition. Dual-labeled microglia/ myeloid mice were used to show myeloid diapedesis. For SAH, 50 μm blood was collected from tail puncture and administered into basal cisterns. IL-6 blockade was given at various time points. Various markers of neuroinflammation were measured with western blot and immunohistochemistry. Cerebral blood flow was also measured. Vasospasm was measured via cardiac injection of India ink/gelatin. Turning test and Garcia's modified SAH score were utilized. P < 0.05 was considered significant.Results: IL-6 expression peaked 3 days following SAH (p < 0.05). Human IL-6 was increased in aneurysmal blood (p < 0.05) and in cerebral spinal fluid (p < 0.01). Receptor upregulation was periventricular and perivascular. Microglia activation following SAH resulted in increased caveolin 3 and myeloid diapedesis. A significant increase in BBB markers endothelin 1 and occludin was noted following SAH, but reduced with IL-6 blockade (p < 0.01). CV occurred 5 days post-SAH, but was absent in IL-6 KO mice and mitigated with IL-6 blockade (p < 0.05). IL-6 blockade, and IL-6 KO mitigated effects of SAH on cerebral blood flow (p < 0.05). SAH mice had impaired performance on turn test and poor modified Garcia scores compared to saline and IL-6 blockade. A distinct microglia phenotype was noted day 5 in the SAH group (overlap coefficients r = 0.96 and r = 0.94) for Arg1 and iNOS, which was altered by IL-6 blockade. Day 7, a significant increase in toll-like receptor 4 and Stat3 was noted. This was mitigated by IL-6 blockade and IL-6 KO, which also reduced Caspase 3 (p < 0.05). To confirm the mechanism, we developed a p-STAT3 inhibitor that targets the IL-6 pathway and this reduced NFΚB, TLR4, and nitrotyrosine (p < 0.001). Ventricular dilation and increased Tunel positivity was noted day 9, but resolved by IL-6 blockade (p < 0.05).

show abstract

“…The development of CRISPR/Cas9 genome editing technologies significantly improved and simplified the development of new transgenic mouse lines [ 353 , 354 , 355 ]. Many lines expressing GEFIs in the brain already exist [ 28 , 30 , 356 , 357 , 358 , 359 , 360 , 361 , 362 ]; in addition, a number of lines express GEFIs in a Cre recombinase-dependent manner and can be bred into distinct Cre recombinase driver mice to direct expression of sensors in the selected cell populations [ 363 ]. Some transgenic lines are commercially available (jax.org).…”

Section: Animal Models For Real-time Imaging Of Cell Signaling Andmentioning

confidence: 99%

Genetically Encoded Tools for Research of Cell Signaling and Metabolism under Brain Hypoxia

et al. 2020

View full text Add to dashboard Cite

Hypoxia is characterized by low oxygen content in the tissues. The central nervous system (CNS) is highly vulnerable to a lack of oxygen. Prolonged hypoxia leads to the death of brain cells, which underlies the development of many pathological conditions. Despite the relevance of the topic, different approaches used to study the molecular mechanisms of hypoxia have many limitations. One promising lead is the use of various genetically encoded tools that allow for the observation of intracellular parameters in living systems. In the first part of this review, we provide the classification of oxygen/hypoxia reporters as well as describe other genetically encoded reporters for various metabolic and redox parameters that could be implemented in hypoxia studies. In the second part, we discuss the advantages and disadvantages of the primary hypoxia model systems and highlight inspiring examples of research in which these experimental settings were combined with genetically encoded reporters.

show abstract

CLARITY analysis of the Cl/pH sensor expression in the brain of transgenic mice

Cited by 7 publications

References 32 publications

DriveLEDs: software for synchronous control and video acquisition of fluorescent signals

DriveLEDs: software for synchronous control and video acquisition of fluorescent signals

Investigation and modulation of interleukin-6 following subarachnoid hemorrhage: targeting inflammatory activation for cerebral vasospasm

Genetically Encoded Tools for Research of Cell Signaling and Metabolism under Brain Hypoxia

Contact Info

Product

Resources

About