Optimized Automated Analysis of Live Neuronal Mitochondria Homeostasis Modulation by Isoform-Specific Retinoic Acid Receptors

Jos&#233; J. M. Vit&#243;ria; Vin&#237;cius de Paula; Odete A. B. da Cruz e Silva; Diogo Trigo

doi:10.3791/65452

Optimized Automated Analysis of Live Neuronal Mitochondria Homeostasis Modulation by Isoform-Spec...

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Neuroscience

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JoVE Journal Neuroscience

Optimized Automated Analysis of Live Neuronal Mitochondria Homeostasis Modulation by Isoform-Specific Retinoic Acid Receptors

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08:33 min

July 28, 2023

DOI: 10.3791/65452-v

José J. M. Vitória¹, Vinícius de Paula¹, Odete A. B. da Cruz e Silva¹, Diogo Trigo¹

¹Neuroscience and Signalling Laboratory, Institute of Biomedicine (iBiMED), Department of Medical Sciences,University of Aveiro

Overview

This study focuses on advancing the analysis of mitochondria, essential organelles for cellular function, by developing automated tools for efficient image processing. Using MATLAB, the researchers addressed the challenges posed by the complex mitochondrial network, enabling rapid analysis of timelapse images. The implications of mitochondrial anomalies for disease mechanisms and therapeutic strategies are also explored.

Key Study Components

Area of Science

Neuroscience
Mitochondrial Biology
Automated Imaging Analysis

Background

Mitochondria are crucial for various cellular functions.
Anomalies in mitochondria are linked to disease processes.
Improved microscopy and computational tools are essential for analyzing mitochondrial function.
High-resolution imaging and bioinformatic strategies are critical for detailed analysis.

Purpose of Study

To develop a MATLAB tool for analyzing live confocal images of mitochondria.
To enhance the efficiency of mitochondrial analysis through automation.
To address challenges in measuring mitochondrial parameters accurately.

Methods Used

Live confocal imaging systems for capturing mitochondrial dynamics.
Automated analysis tools developed using MATLAB.
Machine learning algorithms for image segmentation and quantification.
CRISPR-Cas9 for studying mitochondrial morphology.
Emphasis on developing non-invasive methods for in vivo studies.

Main Results

The study successfully developed a tool that automates mitochondrial analysis.
This tool improves the reliability and efficiency of quantifying mitochondrial parameters.
Addresses the complexity and variability of mitochondrial populations.
Highlights the relevance of mitochondrial function in disease and therapeutic applications.

Conclusions

The developed tool facilitates robust mitochondrial analysis, enhancing research in cell biology and disease mechanisms.
Contributes to the understanding of mitochondria in personalized medicine.
The study underscores the potential for novel therapeutic strategies targeting mitochondrial function.

Frequently Asked Questions

What advantages does the MATLAB tool provide for mitochondrial analysis?

The MATLAB tool automates the analysis process, significantly enhancing efficiency and reducing manual effort in handling large data volumes from mitochondrial imaging.

How can the imaging model be adapted for different experiments?

The imaging model can be adapted by incorporating various types of cell cultures or live imaging systems to study different biological questions concerning mitochondria.

What types of data are obtained through this analysis?

The analysis yields detailed quantification of mitochondrial parameters such as morphology, dynamics, and functional status, contributing to a better understanding of mitochondrial roles.

Are there any limitations to the automated analysis tool?

While the automated tool enhances efficiency, it may require validation against manual measurements to ensure accuracy, especially given mitochondrial heterogeneity.

In what ways can the findings impact therapeutic development?

The findings can lead to insights into mitochondrial-targeted therapies, potentially aiding in the design of personalized medicine approaches focused on mitochondrial dysfunction.

What is the significance of CRISPR-Cas9 in this study?

CRISPR-Cas9 is used to manipulate mitochondrial morphology, providing insights into how alterations at the mitochondrial level affect overall cellular function and health.

The mitochondrial network is extremely complex, making it very challenging to analyze. A novel MATLAB tool analyzes live confocal imaged mitochondria in timelapse images but results in a large output volume requiring individual manual attention. To address this issue, a routine optimization was developed, allowing for speedy file analysis.

My research has focused on the area of neuroscience, and in particular, we want to understand the molecular basis of disease. This has led us to look at cells, their structure, and the organelles that carry out the different cellular functions. Increasingly, mitochondria, they have received more and more attention.

Anomalies at the mitochondrial level are becoming absolutely fundamental to understand disease processes, and they may even provide interesting targets for novel drugs or therapeutic strategies. Here at the University of Aveiro, we have been able to develop tools that are of interest to study mitochondria. Not only microscopy and the more traditional approaches, but also we have been able to optimize automated analysis tools coupled with bioinformatic strategies to be able to quantify and provide more detailed analysis when addressing mitochondrial function.

The most recent advances include the machine learning algorithms for image segmentation and quantification of mitochondrial parameters. And also the high throughput imaging platforms and deep learning algorithms for improving accuracy and efficiency of mitochondrial characterization. Technologies currently used to advance the field of mitochondrial analysis include high resolution microscopy, live imaging systems, computational analysis tools coupled with machine learning protocols, and CRISPR-Cas9 genome editing for mitochondrial morphology and biology analysis.

Current experimental challenges in the field of mitochondria include developing reliable and accurate measurements of mitochondrial parameters, considering the heterogeneity of mitochondrial populations, analyzing the interplay between mitochondrial and cellular biology, and developing non-invasive tools for studying mitochondria in vivo. Our tool enhances efficiency and reliability, allowing for a robust automated analysis of mitochondria. Furthermore, we can explore the potential of hyaluronic acid receptor modulation in personalized medicine for mitochondria.

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