scholarly journals Crosstalk between astrocytes and microglia results in increased degradation of α-synuclein and amyloid-β aggregates

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Jinar Rostami ◽  
Tobias Mothes ◽  
Mahshad Kolahdouzan ◽  
Olle Eriksson ◽  
Mohsen Moslem ◽  
...  
Keyword(s):  
Amyloid Β ◽  
Protein Aggregates ◽  
Time Lapse ◽  
Cell Types ◽  
Alpha Synuclein ◽  
Culture Chamber ◽  
Imaging Data ◽  
Intracellular Protein ◽  
Membrane Structures ◽  
Tunneling Nanotubes

Abstract Background Alzheimer’s disease (AD) and Parkinson’s disease (PD) are characterized by brain accumulation of aggregated amyloid-beta (Aβ) and alpha-synuclein (αSYN), respectively. In order to develop effective therapies, it is crucial to understand how the Aβ/αSYN aggregates can be cleared. Compelling data indicate that neuroinflammatory cells, including astrocytes and microglia, play a central role in the pathogenesis of AD and PD. However, how the interplay between the two cell types affects their clearing capacity and consequently the disease progression remains unclear. Methods The aim of the present study was to investigate in which way glial crosstalk influences αSYN and Aβ pathology, focusing on accumulation and degradation. For this purpose, human-induced pluripotent cell (hiPSC)-derived astrocytes and microglia were exposed to sonicated fibrils of αSYN or Aβ and analyzed over time. The capacity of the two cell types to clear extracellular and intracellular protein aggregates when either cultured separately or in co-culture was studied using immunocytochemistry and ELISA. Moreover, the capacity of cells to interact with and process protein aggregates was tracked using time-lapse microscopy and a customized “close-culture” chamber, in which the apical surfaces of astrocyte and microglia monocultures were separated by a <1 mm space. Results Our data show that intracellular deposits of αSYN and Aβ are significantly reduced in co-cultures of astrocytes and microglia, compared to monocultures of either cell type. Analysis of conditioned medium and imaging data from the “close-culture” chamber experiments indicate that astrocytes secrete a high proportion of their internalized protein aggregates, while microglia do not. Moreover, co-cultured astrocytes and microglia are in constant contact with each other via tunneling nanotubes and other membrane structures. Notably, our live cell imaging data demonstrate that microglia, when attached to the cell membrane of an astrocyte, can attract and clear intracellular protein deposits from the astrocyte. Conclusions Taken together, our data demonstrate the importance of astrocyte and microglia interactions in Aβ/αSYN clearance, highlighting the relevance of glial cellular crosstalk in the progression of AD- and PD-related brain pathology.

scholarly journals Automated analysis of invadopodia dynamics in live cells

2014 ◽  
Author(s):  
Matthew E Berginski ◽  
Sarah J Creed ◽  
Shelly Cochran ◽  
David W Roadcap ◽  
James E Bear ◽  
...  
Keyword(s):  
Protein Complexes ◽  
Metastatic Cancer ◽  
Automated Analysis ◽  
Time Lapse ◽  
Cell Types ◽  
Live Cells ◽  
Imaging Data ◽  
The Matrix ◽  
Invadopodia Formation

Multiple cell types form specialized protein complexes, podosomes or invadopodia and collectively referred to as invadosomes, which are used by the cell to actively degrade the surrounding extracellular matrix. Due to their potential importance in both healthy physiology as well as in pathological conditions such as cancer, the characterization of these structures has been of increasing interest. Following early descriptions of invadopodia, assays were developed which labelled the matrix underneath metastatic cancer cells allowing for the assessment of invadopodia activity in motile cells. However, characterization of invadopodia using these methods has traditionally been done manually with time-consuming and potentially biased quantification methods, limiting the number of experiments and the quantity of data that can be analysed. We have developed a system to automate the segmentation, tracking and quantification of invadopodia in time-lapse fluorescence image sets at both the single invadopodia level and whole cell level. We rigorously tested the ability of the method to detect changes in invadopodia formation and dynamics through the use of well-characterized small molecule inhibitors, with known effects on invadopodia. Our results demonstrate the ability of this analysis method to quantify changes in invadopodia formation from live cell imaging data in a high throughput, automated manner.

scholarly journals Automated analysis of invadopodia dynamics in live cells

2014 ◽  
Author(s):  
Matthew E Berginski ◽  
Sarah J Creed ◽  
Shelly Cochran ◽  
David W Roadcap ◽  
James E Bear ◽  
...  
Keyword(s):  
Protein Complexes ◽  
Metastatic Cancer ◽  
Automated Analysis ◽  
Time Lapse ◽  
Cell Types ◽  
Live Cells ◽  
Imaging Data ◽  
The Matrix ◽  
Invadopodia Formation

Multiple cell types form specialized protein complexes, podosomes or invadopodia and collectively referred to as invadosomes, which are used by the cell to actively degrade the surrounding extracellular matrix. Due to their potential importance in both healthy physiology as well as in pathological conditions such as cancer, the characterization of these structures has been of increasing interest. Following early descriptions of invadopodia, assays were developed which labelled the matrix underneath metastatic cancer cells allowing for the assessment of invadopodia activity in motile cells. However, characterization of invadopodia using these methods has traditionally been done manually with time-consuming and potentially biased quantification methods, limiting the number of experiments and the quantity of data that can be analysed. We have developed a system to automate the segmentation, tracking and quantification of invadopodia in time-lapse fluorescence image sets at both the single invadopodia level and whole cell level. We rigorously tested the ability of the method to detect changes in invadopodia formation and dynamics through the use of well-characterized small molecule inhibitors, with known effects on invadopodia. Our results demonstrate the ability of this analysis method to quantify changes in invadopodia formation from live cell imaging data in a high throughput, automated manner.

Cleavage of Human Embryos: Options and Diversity

Acta Naturae ◽  
2016 ◽  
Vol 8 (3) ◽  
pp. 88-96
Author(s):  
Yu. K. Doronin ◽  
I. V. Senechkin ◽  
L. V. Hilkevich ◽  
M. A. Kurcer
Keyword(s):  
Time Lapse ◽  
Reproductive Technologies ◽  
Human Embryos ◽  
Imaging Data ◽  
Noninvasive Methods ◽  
Topographic Features ◽  
Time Parameters ◽  
Embryo Cleavage ◽  
Time Lapse Imaging ◽  
Developmental Dynamics

In order to estimate the diversity of embryo cleavage relatives to embryo progress (blastocyst formation), time-lapse imaging data of preimplantation human embryo development were used. This retrospective study is focused on the topographic features and time parameters of the cleavages, with particular emphasis on the lengths of cleavage cycles and the genealogy of blastomeres in 2- to 8-cell human embryos. We have found that all 4-cell human embryos have four developmental variants that are based on the sequence of appearance and orientation of cleavage planes during embryo cleavage from 2 to 4 blastomeres. Each variant of cleavage shows a strong correlation with further developmental dynamics of the embryos (different cleavage cycle characteristics as well as lengths of blastomere cycles). An analysis of the sequence of human blastomere divisions allowed us to postulate that the effects of zygotic determinants are eliminated as a result of cleavage, and that, thereafter, blastomeres acquire the ability of own syntheses, regulation, polarization, formation of functional contacts, and, finally, of specific differentiation. This data on the early development of human embryos obtained using noninvasive methods complements and extend our understanding of the embryogenesis of eutherian mammals and may be applied in the practice of reproductive technologies.

scholarly journals Time lapse synchrotron IR chemical imaging for observing the acclimation of a single algal cell to CO2 treatment

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ghazal Azarfar ◽  
Ebrahim Aboualizadeh ◽  
Simona Ratti ◽  
Camilla Olivieri ◽  
Alessandra Norici ◽  
...  
Keyword(s):  
Algal Cell ◽  
Principal Component ◽  
External Perturbation ◽  
Time Lapse ◽  
Chemical Imaging ◽  
Hyperspectral Data ◽  
Aqueous Environment ◽  
Ir Absorption ◽  
Imaging Data ◽  
Mid Infrared

AbstractAlgae are the main primary producers in aquatic environments and therefore of fundamental importance for the global ecosystem. Mid-infrared (IR) microspectroscopy is a non-invasive tool that allows in principle studying chemical composition on a single-cell level. For a long time, however, mid-infrared (IR) imaging of living algal cells in an aqueous environment has been a challenge due to the strong IR absorption of water. In this study, we employed multi-beam synchrotron radiation to measure time-resolved IR hyperspectral images of individual Thalassiosira weissflogii cells in water in the course of acclimation to an abrupt change of CO2 availability (from 390 to 5000 ppm and vice versa) over 75 min. We used a previously developed algorithm to correct sinusoidal interference fringes from IR hyperspectral imaging data. After preprocessing and fringe correction of the hyperspectral data, principal component analysis (PCA) was performed to assess the spatial distribution of organic pools within the algal cells. Through the analysis of 200,000 spectra, we were able to identify compositional modifications associated with CO2 treatment. PCA revealed changes in the carbohydrate pool (1200–950 cm$$^{-1}$$ - 1 ), lipids (1740, 2852, 2922 cm$$^{-1}$$ - 1 ), and nucleic acid (1160 and 1201 cm$$^{-1}$$ - 1 ) as the major response of exposure to elevated CO2 concentrations. Our results show a local metabolism response to this external perturbation.

scholarly journals Brainstem Quadruple Aberrant Hyperphosphorylated Tau, Beta-Amyloid, Alpha-Synuclein and TDP-43 Pathology, Stress and Sleep Behavior Disorders

2021 ◽  
Vol 18 (13) ◽  
pp. 6689
Author(s):  
Lilian Calderón-Garcidueñas ◽  
Ravi Philip Rajkumar ◽  
Elijah W. Stommel ◽  
Randy Kulesza ◽  
Yusra Mansour ◽  
...  
Keyword(s):  
Online Survey ◽  
Amyloid Β ◽  
Alpha Synuclein ◽  
Amyloid Β Peptide ◽  
Hyperphosphorylated Tau ◽  
Post Traumatic Stress ◽  
Impact Of Event Scale ◽  
Sleep Behavior ◽  
Event Scale ◽  
The Impact

Quadruple aberrant hyperphosphorylated tau (p-τ), amyloid-β peptide, alpha-synuclein and TDP-43 brainstem and supratentorial pathology are documented in forensic ≤40y autopsies in Metropolitan Mexico City (MMC), and p-τ is the major aberrant protein. Post-traumatic stress disorder (PTSD) is associated with an elevated risk of subsequent dementia, and rapid eye movement sleep behavior disorder (RBD) is documented in PD, AD, Lewy body dementia and ALS. This study aimed to identify an association between PTSD and potential pRBD in Mexico. An anonymous online survey of 4502 urban college-educated adults, 29.3 ± 10.3 years; MMC, n = 1865; non-MMC, n = 2637, measured PTSD symptoms using the Impact of Event Scale–Revised (IES-R) and pRBD symptoms using the RBD Single-Question. Over 50% of the participants had IES-R scores ≥33 indicating probable PTSD. pRBD was identified in 22.6% of the participants across Mexico and 32.7% in MMC residents with PTSD. MMC subjects with PTSD had an OR 2.6218 [2.5348, 2.7117] of answering yes to the pRBD. PTSD and pRBD were more common in women. This study showed an association between PTSD and pRBD, strengthening the possibility of a connection with misfolded proteinopathies in young urbanites. We need to confirm the RBD diagnosis using an overnight polysomnogram. Mexican women are at high risk for stress and sleep disorders.

Contact behaviour during the reassociation of dissociated epithelial cells in primary culture

1987 ◽  
Vol 88 (4) ◽  
pp. 521-526
Author(s):  
R.M. Brown ◽  
C.A. Middleton
Keyword(s):  
Epithelial Cells ◽  
Chick Embryo ◽  
Primary Culture ◽  
Cell Cultures ◽  
Corneal Epithelium ◽  
Time Lapse ◽  
Cell Types ◽  
Epidermal Cells ◽  
Isolated Cells ◽  
Contact Behaviour

The behaviour in culture of dissociated epithelial cells from chick embryo pigmented retina epithelium (PRE), corneal epithelium (CE) and epidermis has been studied using time-lapse cinematography. The analysis concentrated on the contact behaviour of 60 previously isolated cells of each type during a 24 h period starting 3.5 h after the cells were plated out. During the period analysed the number of isolated cells in cultures of all three types gradually decreased as they became incorporated into islands and sheets of cells. However, there were significant differences in behaviour between the cell types during the establishment of these sheets and islands. In PRE cell cultures, islands of cells developed because, throughout the period of analysis, collisions involving previously isolated cells almost invariably resulted in the development of a stable contact. Once having established contact with another cell these cells rarely broke away again to become reisolated. In contrast the contacts formed between colliding CE and epidermal cells were, at least initially, much less stable and cells of both these types were frequently seen to break away and become reisolated after colliding with other cells. Sheets and islands of cells eventually developed in these cultures because the frequency with which isolated cells become reisolated decreased with increasing time in culture. The possible reasons underlying the different behaviour of PRE cells, when compared with that of CE and epidermal cells, are discussed. It is suggested that the decreasing tendency of isolated CE and epidermal cells to become reisolated may be related to the formation of desmosomes.

scholarly journals ChipSeg: an automatic tool to segment bacteria and mammalian cells cultured in microfluidic devices

2020 ◽  
Author(s):  
Irene de Cesare ◽  
Criseida G. Zamora-Chimal ◽  
Lorena Postiglione ◽  
Mahmoud Khazim ◽  
Elisa Pedone ◽  
...  
Keyword(s):  
Feedback Control ◽  
Mammalian Cells ◽  
Microfluidic Devices ◽  
Time Lapse ◽  
Cell Types ◽  
External Feedback ◽  
Quantitative Measurements ◽  
External Feedback Control ◽  
Automatic Tool ◽  
Cells Cultured

ABSTRACTExtracting quantitative measurements from time-lapse images is necessary in external feedback control applications, where segmentation results are used to inform control algorithms. While such image segmentation applications have been previously reported, there is in the literature a lack of open-source and documented code for the community. We describe ChipSeg, a computational tool to segment bacterial and mammalian cells cultured in microfluidic devices and imaged by time-lapse microscopy. The method is based on thresholding and uses the same core functions for both cell types. It allows to segment individual cells in high cell-density microfluidic devices, to quantify fluorescence protein expression over a time-lapse experiment and to track individual cells. ChipSeg enables robust segmentation in external feedback control experiments and can be easily customised for other experimental settings and research aims.

scholarly journals SPARC Data Structure: Rationale and Design of a FAIR Standard for Biomedical Research Data

2021 ◽  
Author(s):  
Anita Bandrowski ◽  
Jeffrey S. Grethe ◽  
Anna Pilko ◽  
Tom Gillespie ◽  
Gabi Pine ◽  
...  
Keyword(s):  
Data Structure ◽  
Biomedical Research ◽  
Large Scale ◽  
Open Data ◽  
Cell Types ◽  
Research Data ◽  
Imaging Data ◽  
Organ Specific ◽  
The Rich ◽  
Automated Tools

AbstractThe NIH Common Fund’s Stimulating Peripheral Activity to Relieve Conditions (SPARC) initiative is a large-scale program that seeks to accelerate the development of therapeutic devices that modulate electrical activity in nerves to improve organ function. Integral to the SPARC program are the rich anatomical and functional datasets produced by investigators across the SPARC consortium that provide key details about organ-specific circuitry, including structural and functional connectivity, mapping of cell types and molecular profiling. These datasets are provided to the research community through an open data platform, the SPARC Portal. To ensure SPARC datasets are Findable, Accessible, Interoperable and Reusable (FAIR), they are all submitted to the SPARC portal following a standard scheme established by the SPARC Curation Team, called the SPARC Data Structure (SDS). Inspired by the Brain Imaging Data Structure (BIDS), the SDS has been designed to capture the large variety of data generated by SPARC investigators who are coming from all fields of biomedical research. Here we present the rationale and design of the SDS, including a description of the SPARC curation process and the automated tools for complying with the SDS, including the SDS validator and Software to Organize Data Automatically (SODA) for SPARC. The objective is to provide detailed guidelines for anyone desiring to comply with the SDS. Since the SDS are suitable for any type of biomedical research data, it can be adopted by any group desiring to follow the FAIR data principles for managing their data, even outside of the SPARC consortium. Finally, this manuscript provides a foundational framework that can be used by any organization desiring to either adapt the SDS to suit the specific needs of their data or simply desiring to design their own FAIR data sharing scheme from scratch.

scholarly journals Antibiotic action revealed by real-time imaging of the mycobacterial membrane

2022 ◽  
Author(s):  
Michael G. Wuo ◽  
Charles L Dulberger ◽  
Robert A. Brown ◽  
Alexander Sturm ◽  
Eveline Ultee ◽  
...  
Keyword(s):  
Real Time ◽  
Immune Modulation ◽  
Cell Envelope ◽  
Membrane Vesicles ◽  
Time Lapse ◽  
Outer Membrane Vesicles ◽  
Imaging Data ◽  
Minimal Cell ◽  
Mycobacterial Cell ◽  
Cellular Phenotypes

The current understanding of mycobacterial cell envelope remodeling in response to antibiotics is limited. Chemical tools that report on phenotypic changes with minimal cell wall perturbation are critical to gaining insight into this time-dependent phenomenon. Herein we describe a fluorogenic chemical probe that reports on mycobacterial cell envelope assembly in real time. We used time-lapse microscopy to reveal distinct spatial and temporal changes in the mycobacterial membrane upon treatment with frontline antibiotics. Differential antibiotic treatment elicited unique cellular phenotypes, providing a platform for monitoring cell envelope construction and remodeling responses simultaneously. Analysis of the imaging data indicates a role for antibiotic-derived outer membrane vesicles in immune modulation.

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