Validate Mitochondria-Lysosome Contact Site Dysfunction in PD

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Validate Mitochondria-Lysosome Contact Site Dysfunction in PD

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

🧫 Experiment Protocol Validationproposed

SUMMARY

# Validate Mitochondria-Lysosome Contact Site Dysfunction in PD ## Background and Rationale Mitochondria-lysosome contact sites (MCS) represent critical subcellular structures that facilitate organellar communication, lipid transfer, and mitophagy regulation. In Parkinson's disease (PD), mutations in GBA encoding glucocerebrosidase lead to glucosylceramide accumulation within lysosomes, fundamentally disrupting MCS architecture and function. This disruption creates a pathogenic cascade involving

METHODOLOGY NOTES

Phase 1 (Days 1-14): Generate iPSC-derived dopaminergic neurons from GBA-PD patients (n=6 lines) and controls (n=6 lines) using established protocols. Culture cells for 35 days to achieve mature neuronal phenotype. Phase 2 (Days 15-21): Transfect neurons with fluorescent markers (mitoGFP, LysoTracker Red, VPS13D-mCherry) using lipofectamine. Treat experimental groups with glucosylceramide (50μM) or vehicle control for 48h to model GBA dysfunction. Phase 3 (Days 22-25): Perform live-cell confocal microscopy capturing z-stacks every 30 seconds for 10 minutes. Quantify MCS using colocalization analysis (Manders coefficients) and proximity measurements (<150nm distance). Conduct transmission electron microscopy on fixed samples for ultrastructural MCS analysis. Phase 4 (Days 26-28): Execute functional assays including mitochondrial membrane potential (TMRM staining), lysosomal pH (LysoSensor), and glucocerebrosidase activity (4-methylumbelliferyl-β-D-glucopyranoside substrate). Measure alp

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

source	{'type': 'manual', 'source_name': 'wiki', 'extracted_by': 'backfill_v1', 'extraction_date': '2026-04-16T01:00:16.901190Z'}
summary	# Validate Mitochondria-Lysosome Contact Site Dysfunction in PD ## Background and Rationale Mitochondria-lysosome contact sites (MCS) represent critical subcellular structures that facilitate organell
entities	{'genes': ['GBA'], 'diseases': ["Parkinson's Disease"]}
model_system	human
_schema_version	1
experiment_type	validation
primary_outcome	Validate Validate Mitochondria-Lysosome Contact Site Dysfunction in PD
methodology_notes	Phase 1 (Days 1-14): Generate iPSC-derived dopaminergic neurons from GBA-PD patients (n=6 lines) and controls (n=6 lines) using established protocols. Culture cells for 35 days to achieve mature neuro
replication_status	single_study
extraction_metadata	{'backfill_at': '2026-04-16T01:00:16.901195', 'needs_review': True, 'extraction_notes': 'Backfilled from wiki source (no PMID available)', 'extraction_confidence': 0.4}

📊 Evidence Profile Foundational

Evidence Balance

+0%

Certainty

100%

Debates

0

Incoming

718

Outgoing

667

0 supporting 0 contradicting 0 neutral

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Validate Mitochondria-Lysosome Contact Site Dysfunction in PD

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