Analysis of Grayscale Characteristics in Images of Labeled Microtubules from Cultured Cardiac Myocytes

Doublecortin X (DCX), known to be essential for neuronal migration and cortical layering in the developing brain, is a 40 kDa microtubule (MT)-associated protein. DCX directly interacts with MTs via its two structured doublecortin (DC) domains, but the dynamics of this association and the possible regulatory roles played by the flanking unstructured regions remain poorly defined. Here, we employ quantitative fluorescence recovery after photobleaching (FRAP) protocols in living cells to reveal that DCX shows remarkably rapid and complete exchange within the MT network but that the removal of the C-terminal region significantly slows this exchange. We further probed how MT organization or external stimuli could additionally modulate DCX exchange dynamics. MT depolymerisation (nocodazole treatment) or stabilization (taxol treatment) further enhanced DCX exchange rates, however the exchange rates for the C-terminal truncated DCX protein were resistant to the impact of taxol-induced stabilization. Furthermore, in response to a hyperosmotic stress stimulus, DCX exchange dynamics were slowed, and again the C-terminal truncated DCX protein was resistant to the stimulus. Thus, the DCX dynamically associates with MTs in living cells and its C-terminal region plays important roles in the MT-DCX association.

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“…In our study, a higher skewness (Sk > 0) value corresponds to a more heterogeneous distribution of MTs 33 .…”

Section: Methodssupporting

confidence: 50%

Dynamic microtubule association of Doublecortin X (DCX) is regulated by its C-terminus

Moslehi

Bogoyevitch

2017

Sci Rep

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“…We further found that SD and kurtosis based on the histogram of HE staining in the induction group were significantly higher than those of the control group. According to previous reports (20,30,31), SD (or variance or coefficients of variation) and kurtosis derived from the histopathological image were positively associated with frequency and distribution of pixels, which was presumably related to tissue heterogeneity. Therefore, our histopathological results confirmed that a low dose of sorafenib could induce a more heterogeneous HCC compared with HCC without intervention.…”

Section: Discussionmentioning

confidence: 60%

Evaluation of intratumoral heterogeneity by using diffusion kurtosis imaging and stretched exponential diffusion-weighted imaging in an orthotopic hepatocellular carcinoma xenograft model

Guo¹,

Yang²,

Lu³

et al. 2019

Quant. Imaging Med. Surg.

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Background: To investigate the value of diffusion kurtosis imaging (DKI) and diffusion-weighted imaging (DWI) with a stretched exponential model (SEM) in the evaluation of tumor heterogeneity in an orthotopic hepatocellular carcinoma (HCC) xenograft model. Methods: Thirty orthotopic HCC xenograft nude mice models were established and randomly divided into two groups, the sorafenib induction group (n=15) and control group (n=15). Every mouse in each group underwent MRI with DKI and SEM on a 1.5T MR scanner at 7, 14, and 21 days after sorafenib intervention. DKI and SEM parameters including mean kurtosis (MK), mean diffusivity (MD), α, and distributed diffusion coefficient (DDC) were measured, calculated, and compared between the two groups and among different time points. Sequential correlations between histopathological results including necrotic fraction (NF), micro-vessel density (MVD), Ki-67 index, standard deviation (SD), and kurtosis from hematoxylin-eosin staining, and DKI and SEM parameters were analyzed.Results: MK, MD, and DDC of HCC in the sorafenib induction group were significantly higher than those in the control group at each time point (P<0.05), while α was significantly lower (P<0.05). Significantly positive correlations were found between MK and NF (r=0.

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“…In accordance with our previous methods (Dang et al, 2015), microtubule image acquisition was performed using a confocal scanning microscope (TCS SP5; Leica, Wetzlar, Germany) and a 60× oil immersion objective (NA = 1.4). Cy3 was excited at 555 nm, and laser power was controlled at 50%.…”

Section: Imaging and Image Preprocessingmentioning

confidence: 99%

A Quantitative Method for Microtubule Analysis in Fluorescence Images

Lan

et al. 2015

Microsc Microanal

Self Cite

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Microtubule analysis is of significant value for a better understanding of normal and pathological cellular processes. Although immunofluorescence microscopic techniques have proven useful in the study of microtubules, comparative results commonly rely on a descriptive and subjective visual analysis. We developed an objective and quantitative method based on image processing and analysis of fluorescently labeled microtubular patterns in cultured cells. We used a multi-parameter approach by analyzing four quantifiable characteristics to compose our quantitative feature set. Then we interpreted specific changes in the parameters and revealed the contribution of each feature set using principal component analysis. In addition, we verified that different treatment groups could be clearly discriminated using principal components of the multi-parameter model. High predictive accuracy of four commonly used multi-classification methods confirmed our method. These results demonstrated the effectiveness and efficiency of our method in the analysis of microtubules in fluorescence images. Application of the analytical methods presented here provides information concerning the organization and modification of microtubules, and could aid in the further understanding of structural and functional aspects of microtubules under normal and pathological conditions.

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Analysis of Grayscale Characteristics in Images of Labeled Microtubules from Cultured Cardiac Myocytes

Cited by 7 publications

References 22 publications

Dynamic microtubule association of Doublecortin X (DCX) is regulated by its C-terminus

Dynamic microtubule association of Doublecortin X (DCX) is regulated by its C-terminus

Evaluation of intratumoral heterogeneity by using diffusion kurtosis imaging and stretched exponential diffusion-weighted imaging in an orthotopic hepatocellular carcinoma xenograft model

A Quantitative Method for Microtubule Analysis in Fluorescence Images

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