Investigating the Proteins and Mechanisms Involved in Axon Regeneration after Injury

Neurons do not replicate, so they cannot be replaced if they are damaged by neurodegenerative disease, traumatic injury, stroke, etc. This means that the study of how neurons can repair vital structures like axons is very important. Peripheral neurons can regenerate their axons after injury, but central nervous system neurons cannot. The mechanism of peripheral nervous system axon regeneration is not fully understood, so my project focused on determining the proteins involved. Previous studies have shown that the smooth endoplasmic reticulum (ER) must accumulate at the tip of the regenerating axon or axon regeneration will be significantly reduced. I sought to understand why the smooth ER is necessary for axon regeneration by studying the smooth ER calcium channel, ryanodine receptor (RyR). I knocked down expression of RyR using RNAi and compared axon regeneration in knockdown larvae to normal larvae. I found that RyR knockdown significantly reduces axon regeneration after injury which suggests that calcium release from the smooth ER through RyR is involved in axon regeneration. I also tested how RyR knockdown impacts DLK pathway activation after injury and found that it does not significantly affect DLK pathway activation. This suggests that RyR might not be involved in axon regeneration initiation through the DLK pathway. As a whole, my results show that RyR is important for axon regeneration and likely acts somewhere downstream of axon regeneration initiation.

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Work Title Investigating the Proteins and Mechanisms Involved in Axon Regeneration after Injury
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Open Access
Creators
  1. Emily McNaughton
License No Copyright - U.S.
Work Type Research Paper
Acknowledgments
  1. I would like to thank Dr. Melissa Rolls for her support and guidance throughout this project. I also want to thank everyone in the Rolls lab for their kindness and support throughout my project. I especially want to thank Rachel Swope and Ian Hertzler for training me at the beginning of my time in the lab, Michelle Stone for her help on the puc assay, Greg Kothe for his help with the CRISPR work, and Hannah Kline for partnering with me for the beginning of the puc and CRISPR work. Finally, I would like to thank my friends and family for their continued love and support throughout my undergraduate years. This research was funded by the Pennsylvania State University Eberly College of Science, the NASA Pennsylvania Space Grant Consortium Women in Science and Engineering Research program (2020), and National Institute of General Medical Sciences (GM085115). The funders did not contribute to experiment design, data collection, or data analysis.
Publication Date April 26, 2024
Deposited April 26, 2024

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  • Updated Description, Publication Date Show Changes
    Description
    • Neurons do not replicate, so they cannot be replaced if they are damaged by neurodegenerative disease, traumatic injury, stroke, etc. This means that the study of how neurons can repair vital structures like axons is very important. Peripheral neurons can regenerate their axons after injury, but central nervous system neurons cannot. The mechanism of peripheral nervous system axon regeneration is not fully understood, so my project focused on determining the proteins involved. Previous studies have shown that the smooth endoplasmic reticulum (ER) must accumulate at the tip of the regenerating axon or axon regeneration will be significantly reduced. I sought to understand why the smooth ER is necessary for axon regeneration by studying the smooth ER calcium channel, ryanodine receptor (RyR). I knocked down expression of RyR using RNAi and compared axon regeneration in knockdown larvae to normal larvae. I found that RyR knockdown significantly reduces axon regeneration after injury which suggests that calcium release from the smooth ER through RyR is involved in axon regeneration. I also tested how RyR knockdown impacts DLK pathway activation after injury and found that it does not significantly affect DLK pathway activation. This suggests that RyR might not be involved in axon regeneration initiation through the DLK pathway. As a whole, my results show that RyR is important for axon regeneration and likely acts somewhere downstream of axon regeneration initiation.
    Publication Date
    • 2024-04-26
  • Updated Acknowledgments Show Changes
    Acknowledgments
    • I would like to thank Dr. Melissa Rolls for her support and guidance throughout this project. I also want to thank everyone in the Rolls lab for their kindness and support throughout my project. I especially want to thank Rachel Swope and Ian Hertzler for training me at the beginning of my time in the lab, Michelle Stone for her help on the puc assay, Greg Kothe for his help with the CRISPR work, and Hannah Kline for partnering with me for the beginning of the puc and CRISPR work. Finally, I would like to thank my friends and family for their continued love and support throughout my undergraduate years. This research was funded by the Pennsylvania State University Eberly College of Science, the NASA Pennsylvania Space Grant Consortium Women in Science and Engineering Research program (2020), and National Institute of General Medical Sciences (GM085115). The funders did not contribute to experiment design, data collection, or data analysis.
  • Added Creator Emily McNaughton
  • Added Thesis Final Draft.docx
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    • https://rightsstatements.org/page/NoC-US/1.0/
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