Surgical Ablation Assay for Studying Eye Regeneration in Planarians
Autor: | Jacob M. Morton, Marwa A. Saad, Wendy S. Beane |
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Rok vydání: | 2017 |
Předmět: |
Pluripotent Stem Cells
0301 basic medicine genetic structures General Chemical Engineering Population Biology Optic cup (anatomical) Eye Models Biological General Biochemistry Genetics and Molecular Biology 03 medical and health sciences Schmidtea mediterranea Animals Regeneration Photoreceptor Cells education Ocular Physiological Phenomena education.field_of_study General Immunology and Microbiology Regeneration (biology) General Neuroscience Planarians biology.organism_classification eye diseases Nerve Regeneration Eye regeneration 030104 developmental biology Planarian RNA Interference sense organs Stem cell Neuroscience Developmental Biology Adult stem cell |
Zdroj: | Journal of Visualized Experiments. |
ISSN: | 1940-087X |
Popis: | In the study of adult stem cells and regenerative mechanisms, planarian flatworms are a staple in vivo model system. This is due in large part to their abundant pluripotent stem cell population and ability to regenerate all cell and tissue types after injuries that would be catastrophic for most animals. Recently, planarians have gained popularity as a model for eye regeneration. Their ability to regenerate the entire eye (comprised of two tissue types: pigment cells and photoreceptors) allows for the dissection of the mechanisms regulating visual system regeneration. Eye ablation has several advantages over other techniques (such as decapitation or hole punch) for examining eye-specific pathways and mechanisms, the most important of which is that regeneration is largely restricted to eye tissues alone. The purpose of this video article is to demonstrate how to reliably remove the planarian optic cup without disturbing the brain or surrounding tissues. The handling of worms and maintenance of an established colony is also described. This technique uses a 31 G, 5/16-inch insulin needle to surgically scoop out the optic cup of planarians immobilized on a cold plate. This method encompasses both single and double eye ablation, with eyes regenerating within 1-2 weeks, allowing for a wide range of applications. In particular, this ablation technique can be easily combined with pharmacological and genetic (RNA interference) screens for a better understanding of regenerative mechanisms and their evolution, eye stem cells and their maintenance, and phototaxic behavioral responses and their neurological basis. |
Databáze: | OpenAIRE |
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