H9 ES AAVS1-NGN2 FAM134C/A/B-/-
By onES cells modified using CRISPR/Cas9 to lack ER-phagy receptor FAM134C, A & B. Introduced NEUROG2 gene in AAVS1 locus. Derived from human embryonic stem cells at blastocyst stage.
In vivo reduction of age-dependent neuromelanin accumulation mitigates features of Parkinson’s disease
By onHumanized rodents with neuromelanin can develop Parkinson's symptoms. Reducing neuromelanin accumulation mitigates PD features, showing a link between NM levels and disease pathology.
Nucleofection of hPSCs
By onThis protocol describes the standard procedure for the delivery of plasmids, mRNA, or ribonucleoprotein (RNP) into human pluripotent stem cells (hPSCs) using nucleofection.
Thawing of mouse embryonic fibroblasts (MEFs) for hPSC cultures
By onThis protocol describes the thawing of mouse embryonic fibroblasts (MEFs) as feeder cells for human pluripotent stem cell (hPSC) culture.
AAV Purification Protocol with Iodixanol gradient
By onProtocol used in the Kaplitt and Marongiu labs to purify AAVs.
Fluorescent Immunolabelling for Alpha-Synuclein in neuronal primary culture (Testing PFF toxicity)
By onThis protocol is designed to perform fluorescent immunolabelling on neuronal primary culture after PFF incubation. The labelling of phosphorylated alpha-synuclein is considered as a marker of the PFF toxicity and should be done to validate the PFFs.
The calcium sensor synaptotagmin-1 is critical for phasic axonal dopamine release in the striatum and mesencephalon, but is dispensable for basic motor behaviors in mice
By onIn this work, we conditionally deleted the calcium sensor synaptotagmin-1 (Syt1) in DA neurons (cKODA mice) to abrogate most activity-dependent axonal DA release in the striatum and mesencephalon. Syt1 cKODA mice showed intact performance in multiple unconditioned DA-dependent motor tasks, suggesting that activity-dependent DA release is dispensable for such basic motor functions. Basal extracellular levels of DA in the striatum were unchanged, suggesting that a basal tone of extracellular DA is sufficient to sustain basic movement. We also found multiple adaptations in the DA system of cKODA mice, similar to those happening at early stages of PD. Taken together, our findings reveal the striking resilience of DA-dependent motor functions in the context of a near-abolition of phasic DA release, shedding new light on why extensive loss of DA innervation is required to reveal motor dysfunctions in PD.
Non-ablative disease-modifying effects of magnetic resonance-guided focused ultrasound in neuromelanin-producing parkinsonian rodents
By onThe authors' findings indicate that tFUS treatment applied at prodromal/early disease stages provides extended structural and functional preservation of the nigrostriatal pathway in neuromelanin-producing parkinsonian rats.
Statistical analysis
By onThis protocol describes the statistical analysis applied to the quantifications in Chang et al. 2021 SA paper.
Inter-organellar Communication in Parkinson’s and Alzheimer’s Disease: Looking Beyond Endoplasmic Reticulum-Mitochondria Contact Sites
By onNeurodegenerative diseases (NDs) are generally considered proteinopathies but whereas this may initiate disease in familial cases, onset in sporadic diseases may originate from a gradually disrupted organellar homeostasis. Herein, endolysosomal abnormalities, mitochondrial dysfunction, endoplasmic reticulum (ER) stress, and altered lipid metabolism are commonly observed in early preclinical stages of major NDs, including Parkinson's disease (PD) and Alzheimer's disease (AD). Among the multitude of underlying defective molecular mechanisms that have been suggested in the past decades, dysregulation of inter-organellar communication through the so-called membrane contact sites (MCSs) is becoming increasingly apparent. Although MCSs exist between almost every other type of subcellular organelle, to date, most focus has been put on defective communication between the ER and mitochondria in NDs, given these compartments are critical in neuronal survival. Contributions of other MCSs, notably those with endolysosomes and lipid droplets are emerging, supported as well by genetic studies, identifying genes functionally involved in lysosomal homeostasis. In this review, we summarize the molecular identity of the organelle interactome in yeast and mammalian cells, and critically evaluate the evidence supporting the contribution of disturbed MCSs to the general disrupted inter-organellar homeostasis in NDs, taking PD and AD as major examples.
Vibrational Stabilization of Cluster Synchronization in Oscillator Networks
By onRestoring normal cluster synchronization is crucial for system functioning. Vibrational control can stabilize synchronization without state measurements, offering a solution for challenging scenarios.
HeLa ::TMEM192-HA YIPF4-/-
By onHeLa ::TMEM192-HA YIPF4-/- PubMed=37757899; Characteristics: Using CRISPR/Cas9 TMEM192 was C-terminally tagged on both alleles with a 3xHA epitope (from parent cell line). Knockout cell: Method=CRISPR/Cas9; HGNC; 28145; YIPF4. Transformant: NCBI_TaxID; 28285; Adenovirus 5. Derived from site: In situ; Fetal kidney; UBERON=UBERON_0002113. NCBI_TaxID=9606; ! Homo sapiens (Human) RRID:CVCL_C0I5 ! HEK293::TMEM192-3xHA Female Fetus Transformed cell line
Population fraction of Parkinson’s disease attributable to preventable risk factors
By onParkinson's disease is a fast-growing neurologic disease with no known prevention. Environmental factors like head trauma in sports/combat and pesticide exposure contribute significantly to the disease, suggesting preventable causes for some cases.
ER-lysosome lipid transfer protein VPS13C/PARK23 prevents aberrant mtDNA-dependent STING signaling
By onMutations in VPS13C cause early onset, autosomal recessive Parkinson’s Disease (PD). We have established that VPS13C encodes a lipid transfer protein localized to contact sites between the endoplasmic reticulum (ER) and late endosomes/lysosomes. In the current study, we demonstrate that depleting VPS13C in HeLa cells causes an accumulation of lysosomes with an altered lipid profile, including an accumulation of di-22:6-BMP, a biomarker of the PD-associated leucine-rich repeat kinase 2 (LRRK2) G2019S mutation. In addition, the DNA-sensing cGAS/STING pathway, which was recently implicated in PD pathogenesis, is activated in these cells. This activation results from a combination of elevated mitochondrial DNA in the cytosol and a defect in the degradation of activated STING, a lysosome-dependent process. These results suggest a link between ER-lysosome lipid transfer and innate immune activation and place VPS13C in pathways relevant to PD pathogenesis.