Ca2+-Imaging Techniques to Analyze Ca2+ Signaling in Cells and to Monitor Neuronal Activity in the Retina

2012 ◽  
pp. 297-308 ◽  
Author(s):  
Olaf Strauß
Keyword(s):  
Neuronal Activity ◽  
Imaging Techniques ◽  
In Cells

Imaging techniques used for the detection of 8-oxoguanine adducts and DNA repair proteins in cells and tissues

2001 ◽  
Vol 36 (9) ◽  
pp. 1483-1494 ◽  
Author(s):  
Rebecca L. Persinger ◽  
Robert Melamede ◽  
Ivan Bespalov ◽  
Susan Wallace ◽  
Douglas J. Taatjes ◽  
...  
Keyword(s):  
Dna Repair ◽  
Imaging Techniques ◽  
Dna Repair Proteins ◽  
In Cells

Quantitative Fluorescence Imaging Techniques for the Study of Organization and Signaling Mechanisms in Cells

2006 ◽  
pp. 117-141 ◽  
Author(s):  
Margaret H. Wade ◽  
Adriaan W. De Feijter ◽  
Melinda K. Frame ◽  
Melvin Schindler
Keyword(s):  
Fluorescence Imaging ◽  
Imaging Techniques ◽  
Signaling Mechanisms ◽  
Quantitative Fluorescence ◽  
In Cells

Mitochondria and neuronal activity

2007 ◽  
Vol 292 (2) ◽  
pp. C641-C657 ◽  
Author(s):  
Oliver Kann ◽  
Richard Kovács
Keyword(s):  
Energy Metabolism ◽  
Neuronal Activity ◽  
Mitochondrial Function ◽  
Imaging Techniques ◽  
Neurological Diseases ◽  
Experimental Models ◽  
Metabolic Dysfunction ◽  
Membrane Excitability ◽  
Spatiotemporal Resolution ◽  
Atp Production

Mitochondria are central for various cellular processes that include ATP production, intracellular Ca2+ signaling, and generation of reactive oxygen species. Neurons critically depend on mitochondrial function to establish membrane excitability and to execute the complex processes of neurotransmission and plasticity. While much information about mitochondrial properties is available from studies on isolated mitochondria and dissociated cell cultures, less is known about mitochondrial function in intact neurons in brain tissue. However, a detailed description of the interactions between mitochondrial function, energy metabolism, and neuronal activity is crucial for the understanding of the complex physiological behavior of neurons, as well as the pathophysiology of various neurological diseases. The combination of new fluorescence imaging techniques, electrophysiology, and brain slice preparations provides a powerful tool to study mitochondrial function during neuronal activity, with high spatiotemporal resolution. This review summarizes recent findings on mitochondrial Ca2+ transport, mitochondrial membrane potential (ΔΨm), and energy metabolism during neuronal activity. We will first discuss interactions of these parameters for experimental stimulation conditions that can be related to the physiological range. We will then describe how mitochondrial and metabolic dysfunction develops during pathological neuronal activity, focusing on temporal lobe epilepsy and its experimental models. The aim is to illustrate that 1) the structure of the mitochondrial compartment is highly dynamic in neurons, 2) there is a fine-tuned coupling between neuronal activity and mitochondrial function, and 3) mitochondria are of central importance for the complex behavior of neurons.

scholarly journals Direct Evaluation of L-DOPA Actions on Neuronal Activity of Parkinsonian TissueIn Vitro

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Víctor Plata ◽  
Mariana Duhne ◽  
Jesús E. Pérez-Ortega ◽  
Janet Barroso-Flores ◽  
Elvira Galarraga ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Neuronal Activity ◽  
Calcium Imaging ◽  
Neuronal Excitability ◽  
Imaging Techniques ◽  
Direct Evaluation ◽  
Disease Evolution

Physiological and biochemical experimentsin vivoandin vitrohave explored striatal receptor signaling and neuronal excitability to posit pathophysiological models of Parkinson's disease. However, when therapeutic approaches, such as dopamine agonists, need to be evaluated, behavioral tests using animal models of Parkinson's disease are employed. To our knowledge, recordings of population neuronal activityin vitroto assess anti-Parkinsonian drugs and the correlation of circuit dynamics with disease state have only recently been attempted. We have shown that Parkinsonian pathological activity of neuronal striatal circuits can be characterized inin vitrocerebral tissue. Here, we show that calcium imaging techniques, capable of recording dozens of neurons simultaneously with single-cell resolution, can be extended to assess the action of therapeutic drugs. We used L-DOPA as a prototypical anti-Parkinsonian drug to show the efficiency of this proposed bioassay. In a rodent model of early Parkinson's disease, Parkinsonian neuronal activity can be returned to control levels by the bath addition of L-DOPA in a reversible way. This result raises the possibility to use calcium imaging techniques to measure, quantitatively, the actions of anti-Parkinsonian drugs over time and to obtain correlations with disease evolution and behavior.

scholarly journals Quantitative single-protein imaging reveals molecular complex formation of integrin, talin, and kindlin during cell adhesion

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Lisa S. Fischer ◽  
Christoph Klingner ◽  
Thomas Schlichthaerle ◽  
Maximilian T. Strauss ◽  
Ralph Böttcher ◽  
...  
Keyword(s):  
Complex Formation ◽  
Single Molecule ◽  
Molecular Mechanisms ◽  
Detection Efficiency ◽  
Imaging Techniques ◽  
Super Resolution ◽  
Absolute Quantification ◽  
Molecular Probes ◽  
Single Protein ◽  
In Cells

AbstractSingle-molecule localization microscopy (SMLM) enabling the investigation of individual proteins on molecular scales has revolutionized how biological processes are analysed in cells. However, a major limitation of imaging techniques reaching single-protein resolution is the incomplete and often unknown labeling and detection efficiency of the utilized molecular probes. As a result, fundamental processes such as complex formation of distinct molecular species cannot be reliably quantified. Here, we establish a super-resolution microscopy framework, called quantitative single-molecule colocalization analysis (qSMCL), which permits the identification of absolute molecular quantities and thus the investigation of molecular-scale processes inside cells. The method combines multiplexed single-protein resolution imaging, automated cluster detection, in silico data simulation procedures, and widely applicable experimental controls to determine absolute fractions and spatial coordinates of interacting species on a true molecular level, even in highly crowded subcellular structures. The first application of this framework allowed the identification of a long-sought ternary adhesion complex—consisting of talin, kindlin and active β1-integrin—that specifically forms in cell-matrix adhesion sites. Together, the experiments demonstrate that qSMCL allows an absolute quantification of multiplexed SMLM data and thus should be useful for investigating molecular mechanisms underlying numerous processes in cells.

GABA Neurotransmission in the Cerebellar Interposed Nuclei: Involvement in Classically Conditioned Eyeblinks and Neuronal Activity

2004 ◽  
Vol 91 (2) ◽  
pp. 719-727 ◽  
Author(s):  
D. Aksenov ◽  
N. Serdyukova ◽  
K. Irwin ◽  
V. Bracha
Keyword(s):  
Neuronal Activity ◽  
Behavioral Effect ◽  
Tonic Activity ◽  
Neural Responses ◽  
Eyelid Closure ◽  
Cortical Input ◽  
Conditioned Responses ◽  
Classically Conditioned ◽  
In Cells

The cerebellar interposed nuclei (IN) are an essential part of circuits that control classically conditioned eyeblinks in the rabbit. The function of the IN is under the control of GABAergic projections from Purkinje cells of the cerebellar cortex. The exact involvement of cerebellar cortical input into the IN during eyeblink expression is not clear. While it is known that the application of γ-aminobutyric acid-A (GABAA) agonists and antagonists affects the performance of classically conditioned eyeblinks, the effects of these drugs on IN neurons in vivo are not known. The purpose of the present study was to measure the effects of muscimol and picrotoxin on the expression of conditioned eyeblinks and the activity of IN cells simultaneously. Injections of muscimol abolished conditioned responses and either silenced or diminished the activity of IN cells. Two injections were administered in each picrotoxin experiment. The first injection of picrotoxin slightly modified the timing and amplitude of the eyeblink, produced mild tonic eyelid closure, increased tonic activity of IN cells, and reduced the amplitude of the neural responses. The second injection of picrotoxin abolished conditioned responses, further increased tonic eyelid closure, dramatically elevated the tonic activity of IN cells, and in most cases, abolished neuronal responses. These results demonstrate that both GABAA-mediated inactivation and tonic up-regulation of IN cells can interrupt the expression of conditioned eyeblinks and that this behavioral effect is accompanied by the suppression of the neuronal activity correlates of the conditioned stimulus and response.

scholarly journals Mate Choice, Sex Roles and Sexual Cognition: Neuronal Prerequisites Supporting Cognitive Mate Choice

2021 ◽  
Vol 9 ◽  
Author(s):  
Theodora Fuss
Keyword(s):  
Mate Choice ◽  
Neuronal Activity ◽  
Cognitive Abilities ◽  
Behavioral Response ◽  
Sex Roles ◽  
Sex Role ◽  
Imaging Techniques ◽  
Brain Regions ◽  
Ample Evidence ◽  
Differential Coexpression

Across taxa, mate choice is a highly selective process involving both intra- and intersexual selection processes aiming to pass on one’s genes, making mate choice a pivotal tool of sexual selection. Individuals adapt mate choice behavior dynamically in response to environmental and social changes. These changes are perceived sensorily and integrated on a neuronal level, which ultimately leads to an adequate behavioral response. Along with perception and prior to an appropriate behavioral response, the choosing sex has (1) to recognize and discriminate between the prospective mates and (2) to be able to assess and compare their performance in order to make an informed decision. To do so, cognitive processes allow for the simultaneous processing of multiple information from the (in-) animate environment as well as from a variety of both sexual and social (but non-sexual) conspecific cues. Although many behavioral aspects of cognition on one side and of mate choice displays on the other are well understood, the interplay of neuronal mechanisms governing both determinants, i.e., governing cognitive mate choice have been described only vaguely. This review aimed to throw a spotlight on neuronal prerequisites, networks and processes supporting the interaction between mate choice, sex roles and sexual cognition, hence, supporting cognitive mate choice. How does neuronal activity differ between males and females regarding social cognition? Does sex or the respective sex role within the prevailing mating system mirror at a neuronal level? How does cognitive competence affect mate choice? Conversely, how does mate choice affect the cognitive abilities of both sexes? Benefitting from studies using different neuroanatomical techniques such as neuronal activity markers, differential coexpression or candidate gene analyses, modulatory effects of neurotransmitters and hormones, or imaging techniques such as fMRI, there is ample evidence pointing to a reflection of sex and the respective sex role at the neuronal level, at least in individual brain regions. Moreover, this review aims to summarize evidence for cognitive abilities influencing mate choice and vice versa. At the same time, new questions arise centering the complex relationship between neurobiology, cognition and mate choice, which we will perhaps be able to answer with new experimental techniques.

scholarly journals Deciphering spatio-temporal fluorescence changes using multi-threshold event detection (MTED)

2020 ◽  
Author(s):  
Franziska E. Müller ◽  
Volodymyr Cherkas ◽  
Gebhard Stopper ◽  
Laura C. Caudal ◽  
Laura Stopper ◽  
...  
Keyword(s):  
Neuronal Activity ◽  
Event Detection ◽  
Ex Vivo ◽  
Imaging Techniques ◽  
Activity Patterns ◽  
Model Systems ◽  
List Type ◽  
Multiple Thresholds

AbstractRecent achievements in indicator optimization and imaging techniques promote the exploration of Ca2+ activity patterns as a main second messenger in many organs. Astrocytes are important regulators of brain activity and well known for their Ca2+-dependent modulation of neurons. However, standardized methods to analyze and interpret Ca2+ activity recordings are missing and hindering global comparisons. Here, we present a biophysics-based concept to analyze Ca2+signals, which includes multiple thresholds and provides the experimenter with a comprehensive toolbox for a differentiated and in-depth characterization of Ca2+ signals. We analyzed various ex vivo and in vivo imaging datasets and verify the validity of our multi-threshold event detection (MTED) algorithm across Ca2+ indicators, imaging setups, and model systems from primary cell culture to awake, head-fixed mice. Applying our MTED concept enables standardized analysis and advances research using optical readouts of cellular activity to decrypt brain function. It allowed us to obtain new insights into the complex dependence of Ca2+activity patterns on temperature and neuronal activity.Highlights→We present a robust pixel-based algorithm to analyze multidimensional fluorescence data.→Automated multiple-threshold analysis accurately quantifies changes in fluorescence across magnitudes.→It characterizes the complexity of dynamic and overlapping activity patterns of Ca2+ activity of astrocytes in vitro, in situ, and in vivo.→Its application provides quantitative parameters how temperature and neuronal activity determine astrocytic Ca2+ activity.

Cytoplasmic Tubules in Cells Infected with Laryngotracheitis Virus

1969 ◽  
Vol 27 ◽  
pp. 376-377
Author(s):  
A. M. Watrach
Keyword(s):  
Tissue Culture ◽  
Small Proportion ◽  
Chicken Embryo ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Morphologic Characteristics ◽  
Infectious Laryngotracheitis ◽  
Laryngotracheitis Virus ◽  
In Cells

During a study of the development of infectious laryngotracheitis (LT) virus in tissue culture cells, unusual tubular formations were found in the cytoplasm of a small proportion of the affected cells. It is the purpose of this report to describe the morphologic characteristics of the tubules and to discuss their possible association with the development of virus.The source and maintenance of the strain of LT virus have been described. Prior to this study, the virus was passed several times in chicken embryo kidney (CEK) tissue culture cells.

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