Individual cell motility studied by time-lapse video recording: Influence of experimental conditions

Hartmann‐Petersen, Rasmus; Walmod, Peter S.; Berezin, Anton; Berezin, Vladimir; Bock, Elisabeth

doi:10.1002/1097-0320(20000801)40:4<260::aid-cyto2>3.0.co;2-j

Cited by 32 publications

(20 citation statements)

References 38 publications

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“…Many studies of synaptic physiology and plasticity at excitatory synapses have been performed using in vitro brain slices prepared under conditions that involve cooling tissue to temperatures as low as 2°C with electrophysiological measurements made at reduced temperatures. However, studies on cellular dynamics have shown that actin-dependent motility, resulting in significant changes in cell morphology, is strongly attenuated at reduced temperatures, with a Q 10 of ϳ4 ranging from 29 to 39°C (Hartmann-Petersen et al, 2000). Similar values have been recorded for epithelial cells from various teleosts (Ream et al, 2003).…”

Section: Introductionsupporting

confidence: 92%

Hypothermia-Associated Loss of Dendritic Spines

Roelandse¹,

Matus²

2004

J. Neurosci.

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Mechanisms of synaptic plasticity in CNS circuits are commonly investigated using in vitro preparations such as brain slices or slice culture. During their preparation, slices are exposed to low temperatures, and electrophysiological measurements are sometimes made below physiological temperature. Because dendritic spines, which occur at the majority of excitatory synapses, are morphologically plastic, we investigated the influence of reduced temperature on their morphology and plasticity using live cell imaging of hippocampal slices from transgenic mice expressing a green fluorescent protein-based neuronal surface marker and electron microscopy of adult brain slices. Our data show that dendritic spines are highly sensitive to reduced temperature with rapid loss of actin-based motility followed at longer times by reversible loss of the entire spine structure. Thus, reduced temperature significantly affects synaptic morphology, which is in turn known to influence several key aspects of synaptic transmission. Evidence that hypothermia potentiates anesthesia and is associated with spine loss in hibernating animals further suggests that spine morphology may have a widespread influence on brain function.

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Section: Introductionsupporting

confidence: 92%

Hypothermia-Associated Loss of Dendritic Spines

Roelandse¹,

Matus²

2004

J. Neurosci.

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“…Despite the fact that cell motility is being studied intensively and many methodological aspects have been systematically addressed (19), the major impediment in using cell motility as a routine end-point biological assay is the complexity of the required technique combined with the labour-intensive manual processing of thousands of images obtained by time-lapse video recordings.…”

Section: Discussionsupporting

confidence: 59%

An automatic procedure for evaluation of single cell motility

Dmytriyev¹,

Tkach²,

Rudenko³

et al. 2006

Cytometry Pt A

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Background: Cell motility is vital in many physiological and pathological processes, such as embryogenesis, inflammation, wound healing, and metastasis. However, the time-consuming step in the evaluation of individual cell motility is the analysis of hundreds of recorded images of cell cultures in general consisting of retrieving images, one at a time, and marking the positions of individual cells by a computer mouse. Therefore, the aim of the present study was to develop a novel automatic procedure for the evaluation of cell motility. Materials and Methods: The procedure was tested on fibroblasts and glioma and adenocarcinoma cells engineered to express the green fluorescent protein by either transient transfection or adenovirus transduction, allowing automatic recognition of cell coordinates on retrieved images.

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“…Maximal migration speed was found at low collagen concentration, which corresponded to an intermediate level of measured cell attachment strength. This finding is in accord with previous reports in which moderate cell attachment strength favored cell migration due to integrin-substrate interactions [46][47][48]. Moreover, higher ligand densities have been shown to reduce directionality and migration speed [49], which may explain the reduced migratory response of ACL fibroblasts on substrates with higher collagen concentration.…”

Section: Discussionmentioning

confidence: 99%

Effects of Applied DC Electric Field on Ligament Fibroblast Migration and Wound Healing

Chao

Hung

et al. 2007

Connective Tissue Research

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Applied electric fields (static and pulsing) are widely used in orthopedic practices to treat nonunions and spine fusions and have been shown to improve ligament healing in vivo. Few studies, however, have addressed the effect of electric fields (EFs) on ligament fibroblast migration and biosynthesis. In the current study, we applied static and pulsing direct current (DC) EFs to calf anterior cruciate ligament (ACL) fibroblasts. ACL fibroblasts demonstrated enhanced migration speed and perpendicular alignment to the applied EFs. The motility of ligament fibroblasts was further modulated on type I collagen. In addition, type I collagen expression increased in ACL fibroblasts after exposure to pulsing EFs. In vitro wound-healing studies showed inhibitory effects of static EFs, which were alleviated with a pulsing EF. Our results demonstrate that applied EFs augment ACL fibroblast migration and biosynthesis and provide potential mechanisms by which EFs may be used for enhancing ligament healing and repair.

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Individual cell motility studied by time-lapse video recording: Influence of experimental conditions

Cited by 32 publications

References 38 publications

Hypothermia-Associated Loss of Dendritic Spines

Hypothermia-Associated Loss of Dendritic Spines

An automatic procedure for evaluation of single cell motility

Effects of Applied DC Electric Field on Ligament Fibroblast Migration and Wound Healing

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