University Presentation Showcase: Undergraduate Division
Measuring Neurite Degeneration in an In Vitro Model of Parkinson’s Disease using an Automated and Optimized Method
Presenter Hometown
Louisville, KY
Major
Biomedical Sciences
Department
Biological Sciences
Degree
Undergraduate
Mentor
Bradley R. Kraemer
Mentor Department
Biological Sciences
Recommended Citation
Clements, Rachel; Fuller, Lauren; Hall, Wesley; Kalantar, Alborz; and Kraemer, Bradley, "Measuring Neurite Degeneration in an In Vitro Model of Parkinson’s Disease using an Automated and Optimized Method" (2021). University Presentation Showcase Event. 23.
https://encompass.eku.edu/swps/2021/undergraduate/23
Abstract
Neurite degeneration is the process in which the axons and dendrites of a neuron undergo deterioration and is associated with the progression of various neurodegenerative diseases such as Parkinson’s disease (PD). Further studies are needed to understand the molecular factors that underlie neurite degeneration. One commonly used method of quantifying neurite degeneration involves analyzing micrographs using the software FIJI to calculate a Degeneration Index (DI) by determining the ratio of fragmented neurite area to total neurite area. However, this method can be time consuming and subject to inaccuracies. Here we set out to create a more efficient and accurate method of quantifying neurite degeneration. To model neurite degeneration associated with PD, we cultured Lund Human Mesencephalic (LUHMES) cells and induced oxidative stress in culture using 6-hydroxydopamine. Micrographs of degenerating neurites were then used to develop an enhanced method for measuring neurite degeneration. Through optimization of parameters for detecting neurite fragments, enhancing contrast of the neurite images, and using image processing to remove non-neurite materials from the image, we developed a method that calculates a more accurate DI. Additionally, we have created a macro to automate DI measurements, making the method more time efficient and user-friendly than previous methods. To exemplify an application of the improved method, we applied the method in experiments revealing the effects of c-Jun N-terminal Kinase signaling on oxidative stress-induced neurite degeneration. This automated method will allow investigators to more effectively assess neurite degeneration, an important, early-stage event associated with a variety of neurodegenerative diseases.
Presentation format
Poster
Measuring Neurite Degeneration in an In Vitro Model of Parkinson’s Disease using an Automated and Optimized Method
Neurite degeneration is the process in which the axons and dendrites of a neuron undergo deterioration and is associated with the progression of various neurodegenerative diseases such as Parkinson’s disease (PD). Further studies are needed to understand the molecular factors that underlie neurite degeneration. One commonly used method of quantifying neurite degeneration involves analyzing micrographs using the software FIJI to calculate a Degeneration Index (DI) by determining the ratio of fragmented neurite area to total neurite area. However, this method can be time consuming and subject to inaccuracies. Here we set out to create a more efficient and accurate method of quantifying neurite degeneration. To model neurite degeneration associated with PD, we cultured Lund Human Mesencephalic (LUHMES) cells and induced oxidative stress in culture using 6-hydroxydopamine. Micrographs of degenerating neurites were then used to develop an enhanced method for measuring neurite degeneration. Through optimization of parameters for detecting neurite fragments, enhancing contrast of the neurite images, and using image processing to remove non-neurite materials from the image, we developed a method that calculates a more accurate DI. Additionally, we have created a macro to automate DI measurements, making the method more time efficient and user-friendly than previous methods. To exemplify an application of the improved method, we applied the method in experiments revealing the effects of c-Jun N-terminal Kinase signaling on oxidative stress-induced neurite degeneration. This automated method will allow investigators to more effectively assess neurite degeneration, an important, early-stage event associated with a variety of neurodegenerative diseases.
