{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T11:58:53Z","timestamp":1785239933505,"version":"3.55.0"},"reference-count":41,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T00:00:00Z","timestamp":1785196800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Mol. Biosci."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Parkinson\u2019s disease (PD) research traditionally relies on animal models and two-dimensional (2D) culture models. These models fail to recapitulate the complex cellular architecture and network interactions that are possible with three-dimensional (3D) organoid models. While 3D models allow improved cell-cell interactions, spatial organization, and metabolic microenvironments, however, their utility for modelling dopaminergic neurodegeneration remains underexplored.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>We developed and characterized 3D organoids from LUHMES cells which are human embryonic neuronal precursor cells and a well-established human dopaminergic neuronal cell line. We directly compare their responses to the mitochondrial toxin MPP+ against conventional 2D cultures. Functional readouts included cell viability, ATP production, dopaminergic marker expression (tyrosine hydroxylase, MAP2, \u03b2-III tubulin), reactive oxygen species (ROS) generation, electrophysiological activity via multi-electrode arrays (MEA), and calcium signalling dynamics.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>3D LUHMES organoids had 9-fold higher synapsin expression compared to 2D cultures, indicating enhanced synaptic maturity and network complexity. Following acute MPP+ exposure (0.25 mM, 24 h), 3D organoids were significantly more sensitive than 2D cultures, with greater reductions in cell viability (35% vs. 31%), ATP production (36% vs. 31%), and dopaminergic marker expression (TH: 60% reduction in both; MAP2: 70% vs. 54%; TUJ1: 62% vs. 20%). ROS production increased uniformly in 3D organoids (85% positive cells) compared to heterogeneous accumulation in 2D cultures (78% positive cells). Functional assessments revealed that 3D organoids displayed higher baseline electrophysiological activity that was sensitive to impairment following MPP+ treatment, spike amplitude, and calcium signalling responses to various stimuli (ATP, glutamate, GABA).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>LUHMES-derived 3D organoids demonstrate greater physiological relevance for modelling PD-related dopaminergic neurodegeneration than 2D cultures. The enhanced sensitivity to mitochondrial toxins, combined with more sophisticated network architecture and functional properties, suggests this model is a valuable platform for mechanistic studies of neurodegeneration and preclinical drug screening. These findings also support the broader adoption of 3D culture systems in neurodegenerative disease research.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fmolb.2026.1807358","type":"journal-article","created":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T11:09:34Z","timestamp":1785236974000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["LUHMES-derived 3D organoids as an enhanced platform for modelling dopaminergic neurodegeneration in Parkinson's disease"],"prefix":"10.3389","volume":"13","author":[{"given":"Cameron","family":"Keighron","sequence":"first","affiliation":[{"name":"Discipline of Physiology, Cellular Physiology Research Lab, School of Pharmacy and Medical Sciences, University of Galway","place":["Galway, Ireland"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jose C.","family":"Casa-Martinez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Serena","family":"Barral","sequence":"additional","affiliation":[{"name":"Developmental Neurosciences, Zayed Centre for Research, UCL Great Ormond Street Institute of Child Health, University College London","place":["London, United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leo R.","family":"Quinlan","sequence":"additional","affiliation":[{"name":"Discipline of Physiology, Cellular Physiology Research Lab, School of Pharmacy and Medical Sciences, University of Galway","place":["Galway, Ireland"]},{"name":"C\u00daRAM SFI Research Centre for Medical Devices, University of Galway","place":["Galway, Ireland"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,7,28]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"18","DOI":"10.1038\/nrn2759","article-title":"Mechanisms underlying spontaneous patterned activity in developing neural circuits","volume":"11","author":"Blankenship","year":"2010","journal-title":"Nat. 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