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Metabolic Dysfunction as an Indicator of Disease

Mitochondria play central roles in meeting the demands of neuronal synapses for energy (ATP). Mitochondrial dysfunction results in impaired neuroplasticity, neuronal degeneration and cell death, and is now recognized as a key element in neurodegenerative diseases, including Alzheimer’s and Huntington’s diseases, Dementia with Lewy bodies, and Parkinson’s disease. The development of model assay systems such as primary neurons, isolated brain mitochondria and pre-synaptic nerve terminals (synpaptosomes) from specific brain regions have enabled identification of mitochondrial defects associated with neurodegenerative diseases. 

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The Seahorse XF Analyzer has been shown to handle small sample sizes to monitor mitochondrial respiratory parameters of synaptosomes using 50-fold less protein than previously possible.

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A recent announcement by La Trobe University described an exciting step forward in the possibilities of early diagnosis of Parkinson’s Disease. 

Through the bioenergetic analysis of blood cells, this test will enable the detection of abnormal metabolism in the blood cells in people with Parkinson's, opening up the possibility of early treatment for the disease.   Key to this research was the Seahorse Extracellular Flux Analyser, used to analyse the bioenergetic phenotype of cells, and which may be used to identify similar mitochondrial disfunctions in other neurodegenerative conditions such as Huntington’s and Alzheimers disease.  

Label Free time course analysis of neuronal cell behaviour

Neuronal apoptosis and necrosis occur in a large number of neurodegernative diseases.  Even though progress has been made in understanding the principles of cell death signaling in neurons, time course analyses of neuronal cell death had proved to be difficult to achieve.  The xCELLigence System of Real-Time Cell Analyzers developed by ACEA Biosciences enables a non-invasive and label-free way to continuously monitor cellular behavior, and has been utilized to determine the response of neurons to different cell death stimuli, to monitor neuroprotection, and to enable high-throughput screening of anti-Alzheimer’s disease drugs amongst other applications.

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Whole tissue imaging – explore structure and function relationships with X-Clarity

Results utilizing the CLARITY technique have uncovered the 3-D architecture of Alzheimer’s Disease lesions and enabled the study of the distribution of Alzheimer’s disease pathology in 3-D, even in post-mortem human tissue.  Logos Biosystems have further developed the CLARITY technology to make significant improvements for tissue clearing speed, efficiency, and reproducibility.  The X-CLARITY™ Tissue Clearing System from Logos Biosystems is an all-in-one, easy-to-use solution for electrophoretic tissue clearing. Its unique design accelerates the removal of lipids from tissues in a highly efficient manner, producing structurally sound and transparent whole tissues ready for multiple rounds of antibody labeling and imaging. Moreover, the X-CLARITY™ Tissue Clearing System successfully produces transparent tissues that are efficiently penetrated by and labeled with macromolecules such as antibodies or oligonucleotides. This allows for the 3D imaging of large tissues at single-cell resolution.  The method has opened up a world of possibilities, from tracing neural circuitry to exploring the relationship between structure and function. 

The X-CLARITY™ Tissue Clearing System can clear a whole mouse brain in just 6 hours, an astounding 8 times faster than the original technology.

 

Supporting Literature

 ACEA Neurotoxicity

 ACT-PRESTO - Rapid and consistent tissue clearing

 Bioenergetic Analysis of Primary Neuronal Deficiencies 

 ACEA xCELLigence System - Discover what you've been missing

 Seahorse XFp Extracellular Flux Analyzer

 LOGOS X-CLARITY Tissue Clearing System

17.01.2017