1,304 research outputs found
LIPA-α-syn inclusions, but not LIPA-α-syn<sup>ΔNAC</sup> overexpression, disrupt nigrostriatal neuronal transmission.
No full textRepresentative heat maps of Ca2+ signals in one animal overexpressing (A) LIPA-α-syn exposed to blue light, (B) LIPA-α-syn without light stimulation, and (C) LIPA-α-synΔNAC exposed to blue light. Analysis was performed before (Baseline), during (Stimulation day 5), and after (Poststimulation day 10) optogenetic stimulation. The underlying data for (A), (B), and (C) can be found in S1 Data. α-syn, α-synuclein; LIPA, light-inducible protein aggregation. (TIF)</p
LIPA-α-syn aggregation disrupts nigrostriatal neuronal transmission.
No full text(A) Schematic representation of the in vivo experimental paradigm of light-induced α-syn aggregation in the SNc and Ca2+ imaging in the striatum. (B) Confocal microscopy image reconstitution of a sagittal brain slice illustrating the expression of LIPA-α-syn in the SNc and the Ca2+ indicator GCaMP6s in the striatum (n = 5 mice)(scale bar = 500μm). (C) Representative time-lapse live imaging illustration of mini-endoscopic Ca2+ in the striatum. Arrowheads indicate striatal neurons showing Ca2+ activity (scale bar = 20μm). (D) Confocal images illustrating the presence of LIPA-α-syn expression in the majority of dopaminergic TH-positive neurons (n = 5 mice) (scale bar = 20 μm). (E) High magnification of confocal images showing that pathological pS129-positive LIPA-α-syn aggregates persisted in dopaminergic neurons 10 days post-optogenetic stimulation (n = 4–6 mice) (scale bar = 10 μm). (F) Temporal trajectories of synchronicity, frequency, and amplitude of Ca2+ transients before, during, and after light-induced α-syn aggregation (n = 4–5 mice). (G) Temporal trajectories of synchronicity, frequency, and amplitude of Ca2+ transients in the absence of light-induced LIPA-α-syn aggregation (n = 3 mice). (H) Temporal trajectories of synchronicity, frequency, and amplitude of Ca2+ transients before, during, and after light-induction in LIPA-α-synΔNAC (n = 4–5 mice). The data are presented as the means ± SEM. * p ≤ 0.05 and ** p ≤ 0.01. The underlying data for (F) to (H) can be found in S1 Data. α-syn, α-synuclein; LIPA, light-inducible protein aggregation; pS129, phosphorylated α-syn at S129; SNc, substantia nigra pars compacta.</p
LIPA-α-syn aggregation disrupts nigrostriatal transmission.
No full textEndoscopic live imaging of striatal neurons infected with AAV-CAG-GCaMP6s in freely behaving mice before, during, and after induction of LIPA-α-syn aggregates in the midbrain. (MP4)</p
LIPA-α-syn seeds the aggregation of α-syn-GFP.
No full textHEK-293T cells overexpressing LIPA-α-syn and α-syn-GFP constructs and exposed to blue light (456 nm, 0.8 mW/mm2) for 12 hours and fixed in 4% PFA. Z-stack confocal imaging and animation rendering with Imaris software showing the seeding effect of LIPA-α-syn and its coaggregation with α-syn-GFP. (MOV)</p
LIPA-α-syn inclusions recapitulate authentic LB features in cell culture.
No full text(A) Confocal images of HEK-293T cells overexpressing LIPA-α-syn exposed to blue light for 12 hours (0.8 mW/mm2) and stained with LB markers: α-syn (BDlab), phosphorylated α-syn at S129 (pS129), ThS, ubiquitin, HSP70, and p62 (n = 5) (scale bar = 10 μm). (B) Confocal images of hiPSC-derived human neurons overexpressing LIPA-α-syn exposed to blue light for 6 hours (0.4 mW/mm2) and stained with LB markers: α-syn (BDlab), phosphorylated α-syn at S129 (pS129), ThS, ubiquitin, HSP70, p62, and MAP2 (n = 3) (scale bar = 5 μm). (C) Electron micrograph of a cell overexpressing LIPA-α-syn and not exposed to the blue light (scale bar = 2 μm). (D) Electron micrograph of a cell overexpressing LIPA-α-syn and exposed to the blue light for 12 hours. The image illustrates the presence of circular structures (dashed line) corresponding to a LIPA-α-syn inclusion (scale bar = 2 μm). (E) Electron micrograph of a cell overexpressing LIPA-Empty and exposed to the blue light. The arrowhead points to the needle-like LIPA-Empty inclusion (scale bar = 2 μm). (F) At low magnification, LIPA-α-syn inclusion appear to be surrounded by mitochondria and endoplasmic reticulum (yellow arrowhead) and are mainly composed of abundant vesicular structures and dysmorphic organelles (scale bar = 1 μm). (G) At a high magnification, the images reveal the presence of dense core vesicles (yellow asterisk) and dysmorphic mitochondria (blue asterisk) (scale bar = 500 nm). (H) EM images of filamentous structure of α-syn from purified LIPA-α-syn aggregates from HEK-293T cells exposed to blue light for 0, 6, 12 and 24 hours (scale bar 0 hour = 500 nm; scale bar 6, 12, and 24 hours = 200 nm). α-syn, α-synuclein; hiPSC, human-induced pluripotent stem cell; LB, Lewy bodies; LIPA, light-inducible protein aggregation; Nu, nucleus; ThS, thioflavin S.</p
LIPA-α-syn inclusions recapitulate authentic LB features in vivo and precipitate dopaminergic neuronal loss and parkinsonian-like motor impairments.
No full text(A) Experimental design of the experiments using the overexpression and induction of LIPA-α-syn aggregation in the midbrains of WT mice. (B) Confocal microscopy images of representative midbrain neurons with LIPA-α-syn aggregates exhibiting authentic LB markers: α-syn (BDlab), pS129, ThS, ubiquitin, HSP70, and p62 (n = 4 mice) (scale bar = 5 μm). (C) Experimental design of the long-term impact of LIPA-α-syn aggregation on DA neuronal integrity. Treatment with light stimulation (blue lines) was started 2 weeks post-AAV injection, and blue light was applied for 1 hour every other day for 8 weeks. (D) Confocal microscopy images illustrating the LIPA-α-syn or LIPA-Empty inclusions within dopaminergic midbrain neurons. The presence of pathological (pS129-positive) α-syn aggregates was observed only in dopaminergic neurons overexpressing LIPA-α-syn stimulated with blue light (n = 4 mice) (scale bar = 5 μm). (E) Representative confocal microscopy images illustrating dopaminergic neuronal loss in the midbrain of mice overexpressing LIPA constructs and exposed to blue light stimulation (n = 4 mice) (scale bar = 0.5 mm). (F) Stereological quantification of TH-positive dopaminergic neurons and (G) total neuronal markers (Nissl) in the midbrains of mice overexpressing LIPA constructs and exposed (or not exposed) to blue light stimulation. The results are expressed as percentage of the contralateral noninjected side (n = 5–8 mice per experimental condition). The data are presented as the means ± SEM. * p ≤ 0.05, ** p ≤ 0.01, and **** p ≤ 0.0001. Assessment of the behavioral impairment induced by LIPA constructs overexpression with and without blue light stimulation using (H) a grip strength test, (I) cylinder test, (J) rotarod test, and (K) gait test (n = 5–9 mice per experimental condition). The data are presented as the means ± SEM. * p ≤ 0.05, ** p ≤ 0.01, and *** p ≤ 0.0001. The underlying data for (F) to (K) can be found in S1 Data. AAV, adeno-associated virus; α-syn, α-synuclein; LB, Lewy bodies; LIPA, light-inducible protein aggregation; pS129, phosphorylated α-syn at S129; ThS, thioflavin S; WT, wild-type.</p
LIPA-α-syn-induced inclusions recapitulate authentic LB features in the striatum.
No full text(A) Experimental design of the overexpression and induction of LIPA-α-syn aggregation in the striatum of WT mice. (B) Representative confocal microscopy images of striatal neurons with LIPA-α-syn aggregates exhibiting authentic LB markers: α-syn (BDlab and FL140 antibodies), α-syn pS129, thioflavin S, ubiquitin, and p62 (n = 5 mice) (scale bar = 5 μm). (C) Confocal microscopy images of representative striatal neurons overexpressing LIPA-α-syn not exposed to blue light stimulation and stained with authentic LB markers: α-syn (BDlab), pS129, thioflavin S, ubiquitin, HSP70, and p62 (n = 5 mice) (scale bar = 5 μm). AAV, adeno-associated virus; α-syn, α-synuclein; LB, Lewy bodies; LIPA, light-inducible protein aggregation; pS129, phosphorylated α-syn at S129; WT, wild-type. (TIF)</p
LIPA expression, CAD, and genotype.
No full textComparison of LIPA expression in CATHGEN for those with and without CAD based on rs142444 genotype. LIPA exhibits higher expression only in those without CAD in the homozygous minor group (p-value = 0.02).</p
LIPA expression, CAD, and genotype in GTEx.
No full textComparison of LIPA expression in GTEx for those with and without heart disease based on rs142444 genotype. LIPA exhibits higher expression in those without heart disease only in the homozygous minor group (p value = 0.22). (TIF)</p
Assessing multiple regulatory variants for LIPA.
No full textAssessing multiple regulatory variants for LIPA.</p
- …
