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High School Poster

HS Poster

Poster 37 - Can Laser-Induced Shockwave Induce Reactive Astrogliosis?

Saturday, October 26, 2024
10:00 AM - 12:00 PM EST
Location: Exhibit Hall E, F & G

    Co-Author(s)

  • HH

    Huiying Huang, n/a

    Undergraduate student
    University of California, San Diego, United States

  • AL

    Albert Li

    volunteer research intern
    Outreach Program for Advancing Learning in STEM (OPALS), United States

    • Presenting Author(s)

  • MZ

    Mimi Zhao

    volunteer research intern
    Outreach Program for Advancing Learning in STEM (OPALS), United States

    • Co-Author(s)

  • RY

    Ryan Yang

    volunteer research intern
    Outreach Program for Advancing Learning in STEM (OPALS), United States

  • Shawn Kim (she/her/hers)

    volunteer research intern
    Outreach Program for Advancing Learning in STEM (OPALS)
    San Diego, California, United States

  • William Chang

    High School Research Intern
    Outreach Program for Advancing Learning in STEM (OPALS)
    Alhambra, California, United States

  • Yidam Park

    volunteer research intern
    Outreach Program for Advancing Learning in STEM (OPALS)
    San Jose, California, United States

  • CW

    Chengbiao Wu

    Professor
    UCSD School of Medicine Dept of Neuroscience, Outreach Program for Advancing Learning in STEM (OPALS), United States

    • Primary Investigator(s)

  • Linda Shi, PhD (she/her/hers)

    OPALS Program Director, Project Scientist, Co-director IEM
    University of California, San Diego: Institute of Engineering in Medicine
    La Jolla, California, United States

    • Last Author(s)

  • VG

    Veronica Gomez-Godinez

    Scientist
    University of California, San Diego
    La Jolla, California, United States


Introduction: Traumatic brain injury (TBI) is caused when an external blow impacts the cephalic region, causing damage to the central nervous system tissue. Astrocytes, a type of glial cell, respond to injury through reactive astrogliosis. We exposed astrocytes to a laser-induced shockwave (LIS), simulating blast-induced traumatic brain injury [1] [2]. Our experiment studied the protein expression of ionized calcium-binding adaptor molecule 1 (IBA-1) and glial fibrillary acidic protein (GFAP-1) while the cells migrate and change their structures during the healing process. IBA-1 marks microglia, a type of immune cell, which is known to recruit to sites of injury, while GFAP-1 assists in repairing nervous system tissue. With shockwave, the two proteins’ differing expression helps augment understanding of post-TBI reactive astrogliosis and its mechanisms.

Materials and

Methods: Mouse glial cultures isolated from the cortex were subjected to LIS after 93 days in vitro and fixed with 4% paraformaldehyde for staining against GFAP-1 and IBA-1. Entire dishes were imaged and cells were quantified using FIJI/ImageJ and a Python algorithm. The images were stitched together using ImageJ to generate a comprehensive image of the entire plate. Additionally, we analyzed each position individually, determining the cell count and the amount of IBA-1 and GFAP-1 overlap within each field of view (FOV). Three channels were utilized for this task as shown in Figure A, a “blue” channel being DAPI (DNA stain), a “green” channel (IBA-1), and a “red” channel (GFAP-1). Cells that were positive for both IBA-1 and GFAP-1 were categorized as overlapping and quantified.

Results, Conclusions, and Discussions:

Results: The experiment explored expression of IBA-1 and GFAP-1 across all FOVs , using the three color channels. Figure B depicts significant differences between shockwave cells and non-shockwave cells in prominence of IBA-1 (p-value = 0.0144) and GFAP-1 (p-value = 0.0020). Specifically, shockwaved cells exhibited the highest levels of GFAP-1 (52% of cells, 8629/16086 cells), whereas non-shockwave cells had lower levels of GFAP-1 (39% of cells, 1710/4313 cells). Additionally, IBA-1 was more heavily expressed in non-shockwave cells at 13% (549/4313 cells), whereas LIS cells had an expression proportion of 7% (1153/16086 cells). The sample sizes for LIS cells and non-LIS cells were 371 FOVs and 266 FOVs respectively.

Conclusions: In this research, significant differences in protein expression levels were found between shockwave and non-shockwave cells. GFAP-1 expression in LIS cells was markedly higher as shown in Figure B, as the astrocytes must upregulate GFAP-1 to repair injured nervous tissue. The high levels of GFAP-1 may contribute to accelerated healing and negative side effects that accompany it (eg. scarring of brain tissue, expression of immune function in astrocytes). To enhance the reliability of the results, future work should include broadening sample sizes for both shockwave and non-shockwave samples and exploring the effects of Alzheimer’s disease (AD) on reactive astrogliosis. The current method of counting the overlap of GFAP-1 and IBA-1 in cells intoduces selection bias; developing an automated procedure to mitigate bias is crucial.

Acknowledgements (Optional): This project was supported by a generous gift from Beckman Laser Institute Inc. to L.S. and V.G.G. The high school students (*) participated in the UCSD IEM OPALS internship. Dr. Chengbiao Wu provided the astrocytes used in the study. Special thanks to Dr. Shu Chien from UCSD Bioengineering and Dr. Lizhu Chen from CorDx Inc. for their donations.

References (Optional): [1] Veronica Gomez-Godinez, et al. “Laser-Induced Shockwave (LIS) to Study Neuronal Ca2+ Responses”, Front. Bioeng. Biotechnol., 15 February 2021. [2] Pegah Pouladian, et al. “Investigation of Astrocytes’ Morphological Changes in Response to Laser-Induced Shockwave”, BioRxiv, 30 November 2023.