A morphological analysis of growth cones of DRG neurons combining Atomic Force and Confocal Microscopy

Laishram, Jummi; Kondra, Shripad; Avossa, Daniela; Migliorini, Elisa; Lazzarino, Marco; Torre, Vincent

doi:10.1016/j.jsb.2009.09.005

Cited by 19 publications

(12 citation statements)

References 34 publications

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“…These measurements show the significance of the structural properties of actin and tubulin in the growth cone, as the low regions were present primarily in areas devoid of actin or tubulin concentrations. These structures were then confirmed by measurements on living DRGs [21], and later, similar structures were found in the growth cones of living and fixed embryonic stem cell derived neurons [22]. Fluorescence has been combined with AFM to track morphological changes and the destruction of the DRG cytoskeleton upon chemical modification [20].…”

Section: Afm Topography Of Neuronsmentioning

confidence: 82%

Neuron Biomechanics Probed by Atomic Force Microscopy

Spedden

Staii

2013

IJMS

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Mechanical interactions play a key role in many processes associated with neuronal growth and development. Over the last few years there has been significant progress in our understanding of the role played by the substrate stiffness in neuronal growth, of the cell-substrate adhesion forces, of the generation of traction forces during axonal elongation, and of the relationships between the neuron soma elastic properties and its health. The particular capabilities of the Atomic Force Microscope (AFM), such as high spatial resolution, high degree of control over the magnitude and orientation of the applied forces, minimal sample damage, and the ability to image and interact with cells in physiologically relevant conditions make this technique particularly suitable for measuring mechanical properties of living neuronal cells. This article reviews recent advances on using the AFM for studying neuronal biomechanics, provides an overview about the state-of-the-art measurements, and suggests directions for future applications.

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Section: Afm Topography Of Neuronsmentioning

confidence: 82%

Neuron Biomechanics Probed by Atomic Force Microscopy

Spedden

Staii

2013

IJMS

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“…Binary masks of F-actin-and β3-tubulin-positive areas were generated to measure the lamellar area. The lamellar area of stage 1 neurons was defined as the area of F-actin minus the area of β3-tubulin (Laishram et al, 2009). …”

Section: Measurement Of Lamellar Areamentioning

confidence: 99%

Contribution of NADPH-oxidase to the establishment of hippocampal neuronal polarity in culture

Wilson

Núñez

González-Billault

2015

Journal of Cell Science

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Reactive oxygen species (ROS) produced by the NADPH oxidase (NOX) complex play important physiological and pathological roles in neurotransmission and neurodegeneration, respectively. However, the contribution of ROS to the molecular mechanisms involved in neuronal polarity and axon elongation is not well understood. In this work, we found that loss of NOX complex function altered neuronal polarization and decreased axonal length by a mechanism that involves actin cytoskeleton dynamics. These results indicate that physiological levels of ROS produced by the NOX complex modulate hippocampal neuronal polarity and axonal growth in vitro.

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“…Filopodia of hippocampal and DRG GCs have a similar elongated shape with a diameter varying from 80 to 400 nm [31] and an average length of 3.6±0.2 and 5.9±0.6 µm, respectively. In order to measure the force they exert, we positioned a silica bead trapped with an IR laser beam in front of filopodia tips (Figure 2 a and e).…”

Section: Resultsmentioning

confidence: 99%

Comparison of the Force Exerted by Hippocampal and DRG Growth Cones

et al. 2013

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Mechanical properties such as force generation are fundamental for neuronal motility, development and regeneration. We used optical tweezers to compare the force exerted by growth cones (GCs) of neurons from the Peripheral Nervous System (PNS), such as Dorsal Root Ganglia (DRG) neurons, and from the Central Nervous System (CNS) such as hippocampal neurons. Developing GCs from dissociated DRG and hippocampal neurons were obtained from P1-P2 and P10-P12 rats. Comparing their morphology, we observed that the area of GCs of hippocampal neurons was 8-10 µm2 and did not vary between P1-P2 and P10-P12 rats, but GCs of DRG neurons were larger and their area increased from P1-P2 to P10-P12 by 2-4 times. The force exerted by DRG filopodia was in the order of 1-2 pN and never exceeded 5 pN, while hippocampal filopodia exerted a larger force, often in the order of 5 pN. Hippocampal and DRG lamellipodia exerted lateral forces up to 20 pN, but lamellipodia of DRG neurons could exert a vertical force larger than that of hippocampal neurons. Force-velocity relationships (Fv) in both types of neurons had the same qualitative behaviour, consistent with a common autocatalytic model of force generation. These results indicate that molecular mechanisms of force generation of GC from CNS and PNS neurons are similar but the amplitude of generated force is influenced by their cytoskeletal properties.

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A morphological analysis of growth cones of DRG neurons combining Atomic Force and Confocal Microscopy

Cited by 19 publications

References 34 publications

Neuron Biomechanics Probed by Atomic Force Microscopy

Neuron Biomechanics Probed by Atomic Force Microscopy

Contribution of NADPH-oxidase to the establishment of hippocampal neuronal polarity in culture

Comparison of the Force Exerted by Hippocampal and DRG Growth Cones

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