Axon Regeneration In C Elegans
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Axon Regeneration In C Elegans
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Author : Vaclovas Radvilas
language : en
Publisher:
Release Date : 2012
Axon Regeneration In C Elegans written by Vaclovas Radvilas and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2012 with categories.
[ANGLÈS] There exist many illnesses like Alzheimer's disease and stroke or brain injuries that due to degeneration of the nervous system people can not have normal life anymore. This is a consequence of neuron damage which can not recover or regenerate after being severed in adult mammalian brain and spinal cord. On the other hand, there are examples in nature like Caenorhabditis elegans round worm, zebrafish or even mice that their nervous system can recuperate. The aim of this thesis is to design, produce and characterize means of C. elegans immobilization in order to be able to visualize worms' axons and perform laser axotomy. Later, the initial study of spontaneous axon regeneration and possible effect on it after treatment with Citicoline drug (used in patients with mild cerebral ischemia) has to be evaluated. The produced system proved to be sufficient for worm immobilization without making any substantial effects to its health and lifespan. Multiple axotomies were performed and axon regeneration images after 24 hours were acquired using confocal and TPEF microscopy techniques afterwards. Regeneration was visible in 95% of axons in worms that were treated with Citicoline while in control group 90% of axons developed growth cone. Moreover, 33% of regenerating axons connected with the dorsal nerve cord in contrary to 26% in control group. To conclude, this is the first demonstration of possible positive effects to axon regeneration in Citicoline treated C. elegans when microfluidic techniques for immobilization were used for laser axotomy and imaging.
Large Scale Transgenesis And Nerve Regeneration In C Elegans
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Author : Cody Lee Gilleland
language : en
Publisher:
Release Date : 2014
Large Scale Transgenesis And Nerve Regeneration In C Elegans written by Cody Lee Gilleland and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2014 with categories.
Caenorhabditis elegans (C. elegans) is a widely studied model organism due to their fully mapped neural network of 302 neurons and amenable genetics. Their small size and short life cycle allows for rapid studies to be conducted; however, after decades of use the manual methods of manipulation have remained relatively unchanged. This thesis details the development of largely automated, high-throughput, optical, robotic, and microfluidic technologies for laser neurosurgery screening and transgenesis in C. elegans. Discovery of molecular mechanisms and chemical compounds that enhance neuronal regeneration can lead to development of therapeutics to combat nervous system injuries and neurodegenerative diseases. By combining high-throughput microfluidics and femtosecond laser microsurgery, we demonstrate for the first time large-scale in vivo screens for identification of compounds that affect neurite regeneration. We performed over ten thousand microsurgeries at single-axon precision in the nematode C. elegans at a rate of 20 seconds per animal. Following surgeries, we exposed the animals to a hand-curated library of approximately one hundred small-molecules and identified chemicals that significantly alter neurite regeneration. In particular, we found that the PKC kinase inhibitor staurosporine strongly modulates regeneration in a concentration- and neuronal type-specific manner. Two structurally unrelated PKC inhibitors produce similar effects. We further show that regeneration is significantly enhanced by the PKC activator prostratin. Microinjection is an essential and widely used method for C. elegans transgenesis. Traditional injections have remained low-throughput for decades due to the laborious nature of the manual procedure which prohibit large-scale transgenesis screening. This thesis details the development of the C. elegans Automated Micro-Injection (CAMI) system for high-throughput microinjection which could be broadly useful for analysis of gene function through insertion of CRISPR/Cas9, cDNA or RNAi vectors in future applications. Using the standard roller phenotype the transformation efficiency in creating transgenic lines is comparable to manual injections. We demonstrate the utility of the system by deep-phenotyping a subset of 6 mutants backgrounds selected from the million mutant project. We introduced 12 fluorescent reporters to label 38 neurons in each genetic background in order to search for defects in development.
Chemical Biology Studies Of Neuroregenerative Small Molecules Using Caenorhabditis Elegans
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Author : Katherine Hannah Zlotkowski
language : en
Publisher:
Release Date : 2015
Chemical Biology Studies Of Neuroregenerative Small Molecules Using Caenorhabditis Elegans written by Katherine Hannah Zlotkowski and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2015 with categories.
The debilitating effects of spinal cord injury can be attributed to a lack of regeneration in the central nervous system. Identification of growth-promoting pathways, particularly ones that can be controlled by small molecules, could provide significant advancements in regenerative science and lead to potential treatments for spinal cord injury. The biological investigations of neuroregenerative small molecules, specifically the natural products clovanemagnolol and vinaxanthone, have been expanded to a whole organism context using the nematode Caenorhabditis elegans (C. elegans) as a tool for these studies. A straightforward assay using C. elegans was developed to screen for compounds that promote neuronal outgrowth in vivo. This outgrowth assay was then used to guide the design of chemically edited analogs of clovanemagnolol that maintained biological activity while possessing structures amenable to further modification for mechanism of action studies. Pull-down experiments using affinity reagents synthesized from a neuroactive structural derivative, clovanebisphenol, and the C. elegans proteome combined with mass spectrometry-based protein identification and genetic recapitulation using mutant C. elegans identified the putative protein target of the small molecule as a kinesin light chain, KLC-1. Furthermore, the small molecule-promoted regeneration of injured neurons in vivo was studied using laser microsurgery to cut specific axons in C. elegans followed by treatment with a library of analogs of the growth-promoting natural product vinaxanthone. Enhanced axonal regeneration was observed following small molecule treatment and the results were used to determine the structure-activity relationship of vinaxanthone, which may guide future development of potential drug candidates for the treatment of spinal cord injury.
Axon Regeneration
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Author : Ava J. Udvadia
language : en
Publisher: Springer Nature
Release Date : 2023-03-07
Axon Regeneration written by Ava J. Udvadia and has been published by Springer Nature this book supported file pdf, txt, epub, kindle and other format this book has been release on 2023-03-07 with Medical categories.
This volume covers a wide range of approaches utilized to decipher cellular and molecular mechanisms that contribute to successful nerve regeneration leading to functional recovery. Chapters detail a variety of models utilizing both in vivo and in vitro approaches, physical injury models, methods for the isolation/analysis of various macromolecules, live and fixed imaging of regenerating axons, and for quantifying behavioral endpoints enable measurements of regenerative success. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Axon Regeneration: Methods and Protocols aims to be comprehensive guide for researchers.
Parallelized Microfluidic Devices For High Throughput Nerve Regeneration Studies In Caenorhabditis Elegans
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Author : Navid Ghorashian
language : en
Publisher:
Release Date : 2010
Parallelized Microfluidic Devices For High Throughput Nerve Regeneration Studies In Caenorhabditis Elegans written by Navid Ghorashian and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2010 with categories.
The nexus of engineering and molecular biology has given birth to high-throughput technologies that allow biologists and medical scientists to produce previously unattainable amounts of data to better understand the molecular basis of many biological phenomena. Here, we describe the development of an enabling biotechnology, commonly known as microfluidics in the fabrication of high-throughput systems to study nerve degeneration and regeneration in the well-defined model nematode, Caenorhabditis elegans (C. elegans). Our lab previously demonstrated how femtosecond (fs) laser pulses could precisely cut nerve axons in C. elegans, and we observed axonal regeneration in vivo in single worms that were immobilized on anesthetic treated agar pads. We then developed a microfluidic device capable of immobilizing one worm at a time with a deformable membrane to perform these experiments without agar pads or anesthetics. Here, we describe the development of improved microfluidic devices that can trap and immobilize up to 24 individual worms in parallel chambers for high-throughput axotomy and subsequent imaging of nerve regeneration in a single platform. We tested different micro-channel designs and geometries to optimize specific parameters: (1) the initial trapping of a single worm in each immobilization chamber, simultaneously, (2) immobilization of single worms for imaging and fs-laser axotomy, and (3) long term storage of worms on-chip for imaging of regeneration at different time points after the initial axon cut.
Axon Growth And Regeneration Part 1
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Author :
language : en
Publisher: Academic Press
Release Date : 2012-12-31
Axon Growth And Regeneration Part 1 written by and has been published by Academic Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2012-12-31 with Science categories.
Published since 1959, International Review of Neurobiology is a well-known series appealing to neuroscientists, clinicians, psychologists, physiologists, and pharmacologists. Led by an internationally renowned editorial board, this important serial publishes both eclectic volumes made up of timely reviews and thematic volumes that focus on recent progress in a specific area of neurobiology research. This volume reviews existing theories and current research surrounding Axon Growth and Regeneration. Leading authors review state-of-the-art in their field of investigation and provide their views and perspectives for future research Chapters are extensively referenced to provide readers with a comprehensive list of resources on the topics covered All chapters include comprehensive background information and are written in a clear form that is also accessible to the non-specialist
Regulation Of Axon Outgrowth And Guidance In C Elegans
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Author : Jennifer A. Zallen
language : en
Publisher:
Release Date : 1999
Regulation Of Axon Outgrowth And Guidance In C Elegans written by Jennifer A. Zallen and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 1999 with categories.
Axon Growth And Regeneration Part 2
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Author :
language : en
Publisher: Academic Press
Release Date : 2012-12-31
Axon Growth And Regeneration Part 2 written by and has been published by Academic Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2012-12-31 with Medical categories.
Leading authors review state-of-the-art in their field of investigation and provide their views and perspectives for future research. Chapters are extensively referenced to provide readers with a comprehensive list of resources on the topics covered. All chapters include comprehensive background information and are written in a clear form that is also accessible to the non-specialist Leading authors review state-of-the-art in their field of investigation and provide their views and perspectives for future research Chapters are extensively referenced to provide readers with a comprehensive list of resources on the topics covered All chapters include comprehensive background information and are written in a clear form that is also accessible to the non-specialist
The Neurobiology Of C Elegans
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Author :
language : en
Publisher: Elsevier
Release Date : 2006-02-02
The Neurobiology Of C Elegans written by and has been published by Elsevier this book supported file pdf, txt, epub, kindle and other format this book has been release on 2006-02-02 with Science categories.
The Neurobiology of C. elegans assembles together a series of chapters describing the progress researchers have made toward solving some of the major problems in neurobiology with the use of this powerful model organism. The first chapter is an introduction to the anatomy of the C. elegans nervous system. This chapter provides a useful introduction to this system and will help the reader who is less familiar with this system understand the chapters that follow. The next two chapters on learning, conditioning and memory and neuronal specification and differentiation, summarize the current state of the C. elegans field in these two major areas of neurobiology. The remaining chapters describe studies in C. elegans that have provided particularly exciting insights into neurobiology.
Genetic Analysis Of Axon Guidance And Branching In C Elegans
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Author : Joe C. Hao
language : en
Publisher:
Release Date : 2003
Genetic Analysis Of Axon Guidance And Branching In C Elegans written by Joe C. Hao and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2003 with categories.