Automated kymograph analysis for profiling axonal transport of secretory granules

Mukherjee, Amit; Jenkins, Brian V.; Fang, Cheng; Radke, Richard J.; Banker, Gary; Roysam, Badrinath

doi:10.1016/j.media.2010.12.005

Cited by 34 publications

(35 citation statements)

References 31 publications

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“…3 A and B). Kymographs generated from time-lapse images revealed the behavior of individual mitochondria within an axon (50-150 μm in length) (38). Stationary mitochondria appear as vertical lines, and moving mitochondria appear as diagonal lines, with the slope representing the velocity (Fig.…”

Section: Loss Ofmentioning

confidence: 99%

Loss of Miro1-directed mitochondrial movement results in a novel murine model for neuron disease

Nguyen¹,

Weaver

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

192

196

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Defective mitochondrial distribution in neurons is proposed to cause ATP depletion and calcium-buffering deficiencies that compromise cell function. However, it is unclear whether aberrant mitochondrial motility and distribution alone are sufficient to cause neurological disease. Calcium-binding mitochondrial Rho (Miro) GTPases attach mitochondria to motor proteins for anterograde and retrograde transport in neurons. Using two new KO mouse models, we demonstrate that Miro1 is essential for development of cranial motor nuclei required for respiratory control and maintenance of upper motor neurons required for ambulation. Neuron-specific loss of Miro1 causes depletion of mitochondria from corticospinal tract axons and progressive neurological deficits mirroring human upper motor neuron disease. Although Miro1-deficient neurons exhibit defects in retrograde axonal mitochondrial transport, mitochondrial respiratory function continues. Moreover, Miro1 is not essential for calcium-mediated inhibition of mitochondrial movement or mitochondrial calcium buffering. Our findings indicate that defects in mitochondrial motility and distribution are sufficient to cause neurological disease. Motor neuron diseases (MNDs), including ALS and spastic paraplegia (SP), are characterized by the progressive, lengthdependent degeneration of motor neurons, leading to muscle atrophy, paralysis, and, in some cases, premature death. There are both inherited and sporadic forms of MNDs, which can affect upper motor neurons, lower motor neurons, or both. Although the molecular and cellular causes of most MNDs are unknown, many are associated with defects in axonal transport of cellular components required for neuron function and maintenance (1-6).A subset of MNDs is associated with impaired mitochondrial respiration and mitochondrial distribution. This observation has led to the hypothesis that neurodegeneration results from defects in mitochondrial motility and distribution, which, in turn, cause subcellular ATP depletion and interfere with mitochondrial calcium ([Ca 2+ ] m ) buffering at sites of high synaptic activity (reviewed in ref. 7). It is not known, however, whether mitochondrial motility defects are a primary cause or a secondary consequence of MND progression. In addition, it has been difficult to isolate the primary effect of mitochondrial motility defects in MNDs because most mutations that impair mitochondrial motility in neurons also affect transport of other organelles and vesicles (1,(8)(9)(10)(11).In mammals, the movement of neuronal mitochondria between the cell body and the synapse is controlled by adaptors called trafficking kinesin proteins (Trak1 and Trak2) and molecular motors (kinesin heavy chain and dynein), which transport the organelle in the anterograde or retrograde direction along axonal microtubule tracks (7,(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24). Mitochondrial Rho (Miro) GTPase proteins are critical for transport because they are the only known surface receptors that attach mitochondria to...

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Section: Loss Ofmentioning

confidence: 99%

Loss of Miro1-directed mitochondrial movement results in a novel murine model for neuron disease

Nguyen¹,

Weaver

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

192

196

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“…Dysfunctions in this pathway can be detrimental to the cell’s structural integrity and its overall ability to function [4]. Recently, many vesicle classes have been identified that are transported within axons [5]–[7], but their spatial and temporal motility kinetics remain elusive, mainly due to the lack of a suitable model system for such analysis.…”

Section: Introductionmentioning

confidence: 99%

Spatial and Temporal Characteristics of Normal and Perturbed Vesicle Transport

et al. 2014

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Efficient intracellular transport is essential for healthy cellular function and structural integrity, and problems in this pathway can lead to neuronal cell death and disease. To spatially and temporally evaluate how transport defects are initiated, we adapted a primary neuronal culture system from Drosophila larval brains to visualize the movement dynamics of several cargos/organelles along a 90 micron axonal neurite over time. All six vesicles/organelles imaged showed robust bi-directional motility at both day 1 and day 2. Reduction of motor proteins decreased the movement of vesicles/organelles with increased numbers of neurite blocks. Neuronal growth was also perturbed with reduction of motor proteins. Strikingly, we found that all blockages were not fixed, permanent blocks that impeded transport of vesicles as previously thought, but that some blocks were dynamic clusters of vesicles that resolved over time. Taken together, our findings suggest that non-resolving blocks may likely initiate deleterious pathways leading to death and degeneration, while resolving blocks may be benign. Therefore evaluating the spatial and temporal characteristics of vesicle transport has important implications for our understanding of how transport defects can affect other pathways to initiate death and degeneration.

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“…This algorithm is particularly effective for these cases. Regardless of their origins, though, there have been a number of successful efforts made to automate the detection of a particle's path from a kymograph (Mukherjee et al, 2011;Welzel et al, 2009;Zhang et al, 2010) but the methodology to create a kymograph has remained relatively underdeveloped, often at the expense of efficiency and accuracy.…”

Section: Discussionmentioning

confidence: 99%

A novel algorithm to generate kymographs from dynamic axons for the quantitative analysis of axonal transport

Chetta

Shah

2011

Journal of Neuroscience Methods

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Automated kymograph analysis for profiling axonal transport of secretory granules

Cited by 34 publications

References 31 publications

Loss of Miro1-directed mitochondrial movement results in a novel murine model for neuron disease

Loss of Miro1-directed mitochondrial movement results in a novel murine model for neuron disease

Spatial and Temporal Characteristics of Normal and Perturbed Vesicle Transport

A novel algorithm to generate kymographs from dynamic axons for the quantitative analysis of axonal transport

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