Mesencephalic astrocyte-derived neurotrophic factor (MANF) is a recently discovered protein responsible for the regulation and survival of neurons. Studies have shown MANF to be a protective agent that prevents and repairs damaged nervous system tissue, however, how MANF achieves this effect is still poorly understood. We previously found that exposing A. domesticus crickets to a 6 hr 0ºC cold stress leads to numerous proliferative cells in the brain; we hypothesized these are immune cells called hemocytes that enter the brain from the blood to protect brain neurons from damage. Cold-stressed and non-cold-stressed crickets were injected with BrdU and their brains were later removed and processed for anti-BrdU and anti-Ph-H3 immunohistochemistry. Confocal imaging revealed labeled mitotic cells in the presumed location of the blood brain barrier. The cold-stressed crickets were found to have a significantly greater number of these cells. In confocal imaging of blood smears from control adult male crickets, hemocytes labeled for MANF, with anti-MANF immunoreactivity localized to the hemocyte cytoplasm. In our future research, we plan to determine if these new cells may use MANF as part of this protective response. To examine this, we will use immunohistochemistry to compare the presence of MANF in brains of cold stressed crickets and control crickets. Antibodies will be applied to thin sections of brain tissue to fluorescently label cells that contain our target molecule. This allows us to quantify and visualize the MANF-positive cells and their location in different brain regions. Our results will provide further insight into this novel molecule’s role in both the immune and nervous systems. This research has potential implications for a better understanding of neurodegenerative diseases and has provided me with experience with techniques in molecular biology that will benefit my future in the medical field.
Author(s): Anna Shea, Biology and Neuroscience, Abby Benjamin, Biology and Premedical Studies Major
Advisor(s): Kathleen Killian, Department of Biology