Alpha-Synuclein Staging and Spreading in DLB — Spatial Propagation Mapping

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Alpha-Synuclein Staging and Spreading in DLB — Spatial Propagation Mapping

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experiment Created: 2026-04-02T10:01:41 By: crosslink-v2 Quality: 67% ✓ SciDEX ID: experiment-exp-wiki-experiments-dlb-alph

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SUMMARY

# Alpha-Synuclein Staging and Spreading in DLB — Spatial Propagation Mapping ## Background and Rationale Spatial propagation mapping study to define alpha-synuclein spreading patterns in dementia with Lewy bodies (DLB), testing whether cortical-first spreading distinguishes DLB from Parkinson's disease dementia (PDD) where spreading is brainstem-first. **Protocol**: Post-mortem brain mapping in 40 DLB cases and 40 PDD cases across 20 brain regions (Braak staging + cortical and limbic sampling).

METHODOLOGY NOTES

**Phase 1: Cell Line Preparation and Alpha-Synuclein Introduction (Days 1-3)** • Maintain H4 human neuroglioma cells and differentiated SH-SY5Y neuroblastoma cells at 37°C, 5% CO2 • Seed cells in 96-well imaging plates at 5×10^4 cells/well for time-course analysis • Prepare recombinant alpha-synuclein preformed fibrils (PFFs) at 5 μg/mL concentration • Introduce fluorescently-labeled alpha-synuclein (Alexa Fluor 488) to donor cells via lipofection • Establish co-culture system with 1:1 ratio of donor (alpha-synuclein+) to recipient cells **Phase 2: Spatial Propagation Monitoring (Days 4-21)** • Perform live-cell confocal microscopy every 48 hours using 63x oil immersion objective • Track alpha-synuclein aggregation using ThioflavinS staining (1 μg/mL, 30 min incubation) • Measure cell-to-cell transfer using proximity ligation assay (PLA) for alpha-synuclein interactions • Quantify spreading distance using automated image analysis (minimum 200 cells per timepoint) • Apply sequential st

▸Metadatasource: {'type': 'manual', 'source_name': 'wiki'

source	{'type': 'manual', 'source_name': 'wiki', 'extracted_by': 'backfill_v1', 'extraction_date': '2026-04-16T01:00:16.902711Z'}
summary	# Alpha-Synuclein Staging and Spreading in DLB — Spatial Propagation Mapping ## Background and Rationale Spatial propagation mapping study to define alpha-synuclein spreading patterns in dementia with
entities	{'genes': ['DLB'], 'diseases': ["Parkinson's Disease"]}
model_system	cell_line
_schema_version	1
experiment_type	validation
primary_outcome	Regional alpha-synuclein seeding activity map distinguishing DLB from PDD propagation
methodology_notes	Phase 1: Cell Line Preparation and Alpha-Synuclein Introduction (Days 1-3) • Maintain H4 human neuroglioma cells and differentiated SH-SY5Y neuroblastoma cells at 37°C, 5% CO2 • Seed cells in 96-w
replication_status	conflicting
extraction_metadata	{'backfill_at': '2026-04-16T01:00:16.902716', 'needs_review': True, 'extraction_notes': 'Backfilled from wiki source (no PMID available)', 'extraction_confidence': 0.4}

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Alpha-Synuclein Staging and Spreading in DLB — Spatial Propagation Mapping

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