Thesis
Directed generation and functional characterization of human locus coeruleus organoids
Master of Science (M.S.), Drexel University
Jun 2026
DOI:
https://doi.org/10.17918/00011348
Abstract
The locus coeruleus (LC) is a small nucleus located deep within the pons of the brainstem and is the primary source of norepinephrine (NE) within the brain, having diffuse projections throughout the central nervous system. In recent decades, the locus coeruleus has been recognized for its role in neurodegenerative disease, specifically those involving the prion-like spread of toxic protein aggregates throughout the brain, such as frontotemporal dementia, Alzheimer's disease, and supranuclear palsy in the case of tau aggregates, and Parkinson's disease, and dementia with Lewy bodies in the case of alpha-synuclein aggregates. In all of these diseases, the locus coeruleus serves as one of the initial, if not the first, sources of pathology. The relatively recent development of human induced pluripotent stem cell (hiPSC) technologies, specifically 3D region-specific cerebral organoids, has enabled powerful and innovative approaches to model human nervous system development and disease in vitro. However, a human in vitro model of the locus coeruleus remains lacking. This thesis aims to design, optimize, and characterize an organoid model of the human locus coeruleus for disease modeling. In this platform, we are able to reliably generate norepinephrine-enriched locus coeruleus organoids which faithfully recapitulate the developmental hallmarks of the human locus coeruleus in vivo. This was achieved through optimization of Activin A exposure in a time-dependent manner during early neural induction to direct fate-specification of norepinephrine neuron progenitors. Model fidelity was validated using extensive RT-qPCRs and immunofluorescence analyses confirm the presence, abundance, and spatial organization of LC-specific markers, by comparison to cortical organoids and parental hiPSC lines. Functional maturation was assessed using multielectrode array (MEA) recordings to quantify spontaneous and network-level electrophysiological activity. Moreover, organoids virally transduced with genetically encoded norepinephrine sensors enabled real-time detection of norepinephrine release and signaling dynamics, demonstrating robust noradrenergic functionality. Together, these data indicate that the model can recapitulate some key developmental, cellular, and molecular features of the human locus coeruleus and establishes a physiologically relevant platform for investigating neurodevelopmental processes and neurodegenerative disease mechanisms.
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Details
- Title
- Directed generation and functional characterization of human locus coeruleus organoids
- Creators
- Gabriel Vuotto Paschall
- Contributors
- Kimberly J. Dougherty (Advisor)
- Awarding Institution
- Drexel University
- Degree Awarded
- Master of Science (M.S.)
- Publisher
- Drexel University
- Number of pages
- viii, 57 pages
- Resource Type
- Thesis
- Language
- English
- Academic Unit
- College of Medicine; Neurology; Drexel University
- Other Identifier
- 991022178035404721