Speaker: Aashu Sheeti, PhD Postdoctoral scholar in the lab of Murali Parkiya
Title: "Orai2 negatively regulates SOCE and pro-inflammatory phenotype in astrocytes "
Abstract: Astrocytes are the most abundant glial cells in the brain and regulate essential functions such as synapse development, blood flow, and the maintenance of neurotransmitter and ion concentrations in the extracellular space. Astrocytes also undergo reactive transformation, leading to the loss of their neurotrophic functions and the release of cytokines and chemokines that contribute to neuroinflammation. Our previous studies identified store-operated calcium entry (SOCE) as a key regulator of astrocyte function. Additionally, deletion of the Ca²⁺ channel Orai1 results in the loss of SOCE and reduces the reactive astrocytic phenotype. Currently, my work focuses on understanding the role of the Orai1 homolog, Orai2, in astrocytes. Deletion of Orai2 in astrocytes leads to elevated SOCE when stimulated by purinergic agonists (ATP and UTP) and the protease activated receptor ligand thrombin. Furthermore, SOCE-induced cytokine expression is increased in Orai2-deficient astrocytes compared to wild-type astrocytes. These findings suggest that Orai2 is a key regulator of the astrocyte transition to a pro-inflammatory state, and the ratio of Orai1/Orai2 dictates the magnitude of SOCE and it’s impact on astrocyte effector function. Future studies will evaluate its role in regulating gliotransmitter release and astrocyte metabolism.
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Speaker: Viridana Leon PhD Candidate in the lab of Joshua Ziarek
Title: "Development of Novel Isotope Labeling Method for Rapid NMR Resonance Assignment"
Abstract: Approximately 10-25% of the human proteome is composed of intrinsically disordered proteins (IDPs), but rational drug discovery and structural studies are impeded by their dynamic nature. We use nuclear magnetic resonance (NMR) spectroscopy to capture their conformational ensembles and dynamic behavior in solution, but spectral overlap and limited resolution pose challenges for disordered proteins. We are developing a new method called codon-specific isotope labeling, to solve this problem. This approach employs cell free protein synthesis to selectively assign unique isotopologues (e.g. 15N, 15N/13C, etc.) to each codon. Each isotopologue produces a unique NMR spectral pattern (e.g. singlet, doublet, etc.) that facilitates rapid assignments. It will apply to an array of biomolecules compatible with cell free expression systems.
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