Investigating Functional Connectivity in a TBCK Loss-of-Function Human Induced Neuronal Model

Presenter

Ethan Howell

Campus

UMass Amherst

Sponsor

ChangHui Pak, Department of Biochemistry and Molecular Biology, UMass Amherst

Schedule

Session 5, 3:30 PM - 4:15 PM [Schedule by Time][Poster Grid for Time/Location]

Location

Poster Board A47, Campus Center Auditorium, Row 3 (A41-A60) [Poster Location Map]

Abstract

First identified in 2016, TBCK syndrome involves a biallelic deletion of the TBCK gene. The TBCK protein is essential for stabilization of intracellular trafficking and dysfunction in TBCK has been tied to issues in mTOR signaling, lysosomal storage, early endosomal trafficking, and mitochondrial respiration. The TBCK mutation has yet to be fully characterized in induced neuronal cultures, and information surrounding the connectivity and morphology of TBCK loss of function neurons could elucidate the mechanisms  through which symptoms such as epilepsy, hypotonia, and intellectual disability arise. To study the effects of TBCK syndrome in neurons, we employed CRISPR-edited induced pluripotent stem cells to be differentiated into neuronal cell cultures. Using high-density microelectrode arrays, we measured action potentials emitted by TBCK knock-out neurons and assessed the synchronization in comparison to control cultures. To further evaluate functionality, neuronal morphology and early endosomal localization in GFP transfected and RFP infected cells was measured through live imaging. Preliminary results have shown significant decreases in the amplitude and firing rates of action potentials in mutant cultures which hints at reduced synchronous activity in TBCK knock-out neurons. A deeper characterization of the effect of TBCK syndrome on neuronal functionality will set a foundation for further therapeutic advances in the disease and provide insight into similar neurodevelopmental disorders.

Keywords

Neurodevelopmental Disorders, TBCK Syndrome, Neuronal Connectivity, Induced Pluripotent Stem Cells, Cell Morphology

Research Area

Neuroscience and Cognitive Science

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