full length human α syn  (Addgene inc)

Journal: Journal of Advanced Research

Article Title: Calcium influx: An essential process by which α-Synuclein regulates morphology of erythrocytes

doi: 10.1016/j.jare.2023.09.009

Figure Lengend Snippet: Morphological abnormalities of RBCs in PD patients accompanied by increased membrane-bound aggregated α-syn. ( A - E ) Representative RBC images of healthy controls (HC) ( A ), patients with Parkinson's disease (PD) ( B ), patients with Alzheimer’s disease (AD) ( C ), patients with mild cognitive impairment (MCI) ( D ), and patients with essential tremor patients (ET) ( E ). Scale bar: 3 μm. ( F ) The percentage of normally-shaped RBCs in the PD group was significantly different from that in the HC group. N = 1000 randomly selected RBCs counted from each individual. Statistical significance was calculated on the percentage of normal RBCs between patients and HC via one-way ANOVA followed by Tukey's post-hoc test. ( G ) The level of total α-syn in the cytosolic and membrane fractions of RBCs was examined in PD and HC. ( H ) The level of aggregated α-syn in the RBC cytosolic and membrane fractions was examined in PD and HC. Values are means ± S.E.M; ns, not significant; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: Briefly, 1 μg/μl full-length monomeric α-syn (12093-HNAE, Sino Biological) was incubated at 37 °C with shaking using Multiscan GO (Thermo Fisher) for 120 h.

Techniques: Membrane

Journal: Journal of Advanced Research

Article Title: Calcium influx: An essential process by which α-Synuclein regulates morphology of erythrocytes

doi: 10.1016/j.jare.2023.09.009

Figure Lengend Snippet: Morphological abnormalities of RBCs and the expression of α-syn in A53T mice and SNCA-KO mice. (A) Scanning electron micrographs of RBCs of WT mice and A53T mice. Scale bar: 1 μm. (B) Proportion of normal RBCs in WT mice and A53T mice. (C) Proportion of acanthocytes in WT mice and A53T mice. N = 5 mice, 1000 randomly selected RBCs counted from each mouse. (D) The level of aggregated α-syn was examined in WT mice and A53T mice (N = 5). (E) Representative western blot images of α-syn in SNCA-KO mice. (F) Scanning electron micrograph showing the morphology of SNCA-KO RBCs. Scale bar: 500 nm. (G) No significant difference was observed in the proportion of RBCs with normal morphology in WT and SNCA-KO mice. (H) The proportion of acanthocytes in WT mice was significantly higher than in SNCA-KO mice. N = 8 mice, 1000 randomly selected RBCs counted for each mouse. Values are presented as means ± S.E.M; ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: Briefly, 1 μg/μl full-length monomeric α-syn (12093-HNAE, Sino Biological) was incubated at 37 °C with shaking using Multiscan GO (Thermo Fisher) for 120 h.

Techniques: Expressing, Western Blot

Journal: Journal of Advanced Research

Article Title: Calcium influx: An essential process by which α-Synuclein regulates morphology of erythrocytes

doi: 10.1016/j.jare.2023.09.009

Figure Lengend Snippet: The effect of α-syn on the morphology of RBCs. (A–C) Scanning electron micrographs show RBCs of WT mice ( A ), SNCA-KO mice ( B ), and A53T mice ( C ) treated with different concentrations of monomeric α-syn (α-syn) or aggregated α-syn (α-syn aggregates) for 48 h. Scale bar: 1 μm. (D–F) Comparison of the percentage of RBCs in WT mice ( D ), SNCA-KO mice ( E ), and A53T mice ( F ) treated with α-syn. N = 5 mice, 1000 randomly selected RBCs counted for each mouse. Statistical significance was calculated on the percentage of normal RBCs between certain groups via Student’s t -test. Values are presented as means ± S.E.M; ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: Briefly, 1 μg/μl full-length monomeric α-syn (12093-HNAE, Sino Biological) was incubated at 37 °C with shaking using Multiscan GO (Thermo Fisher) for 120 h.

Techniques: Comparison

Journal: Journal of Advanced Research

Article Title: Calcium influx: An essential process by which α-Synuclein regulates morphology of erythrocytes

doi: 10.1016/j.jare.2023.09.009

Figure Lengend Snippet: The level of aggregated α-syn on the membrane of mouse RBCs might affect the RBC morphology. (A) RT-QuIC responses observed with reactions seeded with RBC samples of mice treated with PHZ. (B) The RBC morphology in blood samples of control group over time. Scale bar: 1 μm. ( C ) The RBC morphology in PHZ treatment group over time. (D) Comparison of the proportion of RBCs with normal morphology in control and PHZ treated mice (N = 6 mice, 1000 randomly selected RBCs counted for each mouse). (E) Comparison of the proportion of acanthocytes in control and PHZ treated mice (N = 6 mice, 1000 randomly selected RBCs counted for each). Values are presented as means ± S.E.M; ns, not significant; *, P < 0.05.

Article Snippet: Briefly, 1 μg/μl full-length monomeric α-syn (12093-HNAE, Sino Biological) was incubated at 37 °C with shaking using Multiscan GO (Thermo Fisher) for 120 h.

Techniques: Membrane, Control, Comparison

Journal: Journal of Advanced Research

Article Title: Calcium influx: An essential process by which α-Synuclein regulates morphology of erythrocytes

doi: 10.1016/j.jare.2023.09.009

Figure Lengend Snippet: Aggregated α-syn induced intracellular calcium increase via influx of extracellular calcium. (A) Representative confocal images showing RBCs of WT mice in response to different concentrations of monomeric α-syn at the times indicated during the trace. Scale: 1 μm. (B) Traces show calcium dependent fluorescence of RBCs of WT mice in response to different concentrations of monomeric α-syn. N = 20 RBCs from 3 biologically independent experiments. (C) Representative confocal images showing RBCs of WT mice in response to different concentrations of aggregated α-syn at the times indicated during the trace. Scale bar: 1 μm. (D) Traces show calcium-dependent fluorescence of RBCs of WT mice in response to different concentrations of aggregated α-syn. N = 20 RBCs from 3 biologically independent experiments. (E) Representative confocal images showing RBCs of WT mice in response to 0.75 μg/ml aggregated α-syn with PBS as control, with or without Amlodipine, at the time points indicated during the trace. Scale bar: 1 μm. (F) Traces show calcium dependent fluorescence of RBCs of WT mice in response to 0.75 μg/ml aggregated α-syn, with or without Amlodipine. N = 20 RBCs from 3 biologically independent experiments. (G) Scanning electron micrographs showing RBCs of WT mice treated with 0.75 μg/ml monomeric α-syn (α-syn) and aggregated α-syn (α-syn aggregates), with or without Amlodipine. Scale bar: 2 μm. (H) Comparison of the percentage of RBCs of WT mice treated with 0.75 μg/ml monomeric α-syn and aggregated α-syn, with or without Amlodipine (N = 1000). Statistical significance was determined for the percentage of normal RBCs between certain groups via Student’s t -test. Values are means ± S.E.M; ns, not significant; *, P < 0.05.

Article Snippet: Briefly, 1 μg/μl full-length monomeric α-syn (12093-HNAE, Sino Biological) was incubated at 37 °C with shaking using Multiscan GO (Thermo Fisher) for 120 h.

Techniques: Fluorescence, Control, Comparison