A strategy to assess the effect of microgravity on the neuronal cytoskeleton and intracellular trafficking

Microgravity exposure plays a role in the onset of physiological dysfunction among crew onboard the International Space Station. Cognitive effects including impaired motor control and mood disturbance are difficult for astronauts, as they decrease quality of life and are risk factors for neurological conditions. However, it remains difficult to investigate the underlying cause of such conditions via cellular experimentation in actual microgravity. Here, in collaboration with HNu Photonics (Maui, HI), I report a framework for use of the imaging and microfluidics platform Mobile SpaceLab™ for cell biology research in microgravity. We plan to send SH-SY5Y neuroblastoma cells cultured in the MoSL to the ISS (SpaceX27, Jan.2023) and will investigate the effect of microgravity on neurite outgrowth, microtubule dynamics and axonal transport of mitochondria. In this thesis, I establish the culture and imaging conditions for the ground control, a data set which will serve as a negative control for the effect of microgravity on SH-SY5Y cells sent to outer space. Furthermore, I establish and validate protocols for post-flight analysis; a regimen of immunostaining and quantitative analyses to identify cell stress and mitochondrial dysfunction post-microgravity exposure. The data collected in this thesis will allow for analysis of the first representations of neurite outgrowth dynamics and axonal transport in actual microgravity conditions. These results may aid in uncovering mechanistic pathways involving microtubule disruption and help explain maladaptive responses to microgravity in neurons. Furthermore, success in this mission may open the hatch for follow-up experiments using this framework and MoSL across cell biology.