Journal: bioRxiv

Article Title: Nanoscale CaV channel reorganization links α-synuclein pathology to calcium-dependent transcriptional dysregulation

doi: 10.64898/2026.05.01.719272

Figure Lengend Snippet: A Schematic illustrating imaging at the plasma membrane (PM). B Left: representative single-plane Airyscan confocal images of the PM showing Ca V 1.2 immunolabeling in control (CTL, black) and PFF-treated (red) neurons. Inset: MAP2 (pink) neuronal marker. Right: quantification of Ca V 1.2 cluster size, cluster density, and mean gray value (MGV) in the soma (a.) and dendrites (b.) of CTL (black) and PFF-treated (red) neurons. Dendritic measurements are shown separately for excitatory (dark blue) and inhibitory (light blue) populations. n = 20 somata per condition; n = 20 dendrites per group (CTL excitatory, CTL inhibitory, PFF excitatory, PFF inhibitory); two independent isolations with each isolation containing 8-10 pups. C Left: representative super-resolution TIRF localization maps showing Ca V 1.2 immunolabeling in CTL (black) and PFF-treated (red) neurons. Right: quantification of PM Ca V 1.2 cluster size, cluster density, and nearest-neighbor distance in the somatic region. n = 16 neurons per condition; two independent isolations. D Same experimental design as in ( B ), with neurons immunolabeled for Ca V 2.1. n = 19 (CTL) and n = 20 (PFF) somata; n = 20 dendrites per group (CTL excitatory, CTL inhibitory, PFF excitatory, PFF inhibitory); two independent isolations. E Same experimental design as in ( C ), with neurons immunolabeled for Ca V 2.1. n = 16 neurons per condition; two independent isolations. Error bars represent SEM. Statistical significance was determined using two-tailed Mann-Whitney or unpaired two-tailed t-tests. ns, not significant; *P ≤ 0.05; ***P ≤ 0.001; ****P ≤ 0.0001. CTL, control; PFF, α-synuclein pre-formed fibril treatment.

Article Snippet: For negative controls, Ca V 1.2 antibody (Alomone, ACC-003) was applied alone.

Techniques: Imaging, Clinical Proteomics, Membrane, Immunolabeling, Control, Marker, Isolation, Two Tailed Test, MANN-WHITNEY

Journal: bioRxiv

Article Title: Nanoscale CaV channel reorganization links α-synuclein pathology to calcium-dependent transcriptional dysregulation

doi: 10.64898/2026.05.01.719272

Figure Lengend Snippet: A Schematic representation of the CCAD peptide mechanism of action. B Left: representative single-plane Airyscan confocal images of the PM in CTL and PFF-treated neurons co-incubated with SCRBL or CCAD peptides and co-immunolabeled for Ca V 1.2 and K V 2.1. Conditions are shown as CTL;SCRBL (black), PFF;SCRBL (red), CTL;CCAD (gray), and PFF;CCAD (yellow). Right: quantification of Ca V 1.2 cluster size, K V 2.1 cluster size, and Ca V 1.2-K V 2.1 overlap area in the somatic region. n = 19 (CTL;SCRBL), n = 19 (PFF;SCRBL), n = 20 (CTL;CCAD), and n = 20 (PFF;CCAD) neurons; two independent isolations. C Schematic representation of the proximity ligation assay (PLA). D Top: representative Airyscan confocal PLA images showing Ca V 1.2-K V 2.1 proximity. Images are maximum intensity projections from Z-stacks spanning whole cells. Color coding as in ( B ). Bottom: quantification of PLA puncta density. n = 18 neurons per condition; two independent isolations. E Same experimental design as in ( D ), but assessing Ca V 2.1-K V 2.1 proximity. n = 18 neurons per condition; two independent isolations. Error bars represent SEM. Statistical significance in panels ( B, D-E ) was determined using two-way ANOVA with appropriate post hoc tests. ns, not significant; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001. CTL, control; PFF, α-synuclein pre-formed fibril treatment; CCAD, calcium channel association domain peptide; SCRBL, scrambled control peptide.

Article Snippet: For negative controls, Ca V 1.2 antibody (Alomone, ACC-003) was applied alone.

Techniques: Incubation, Immunolabeling, Proximity Ligation Assay, Control

Journal: bioRxiv

Article Title: Nanoscale CaV channel reorganization links α-synuclein pathology to calcium-dependent transcriptional dysregulation

doi: 10.64898/2026.05.01.719272

Figure Lengend Snippet: A Left: representative FV4000 confocal images showing co-immunolabeling of CDK5 and K V 2.1 at the PM in control (CTL, black) and PFF-treated (red) neurons. Images are maximum intensity projections from three optical sections acquired at the PM. Right: quantification of CDK5-K V 2.1 overlap area and CDK5 puncta density in the somatic region. n = 18 (CTL) and n = 19 (PFF) neurons; two independent isolations. B Schematic representation of the roscovitine mechanism of action. C Top: representative FV4000 confocal images of the PM in CTL and PFF-treated neurons incubated with or without roscovitine and immunolabeled for pS603-K V 2.1. Conditions are shown as CTL (black), PFF (red), CTL;Rosco (blue), and PFF;Rosco (purple). Images are maximum intensity projections from Z-stacks spanning whole cells. Bottom: quantification of somatic pS603-Kv2.1 occupancy (% of soma area), cluster MGV, and cluster density. n = 19 (CTL), n = 20 (PFF), n = 20 (CTL;Rosco), and n = 20 (PFF;Rosco) neurons; two independent isolations. D Left: representative FV4000 confocal PLA images showing Ca V 1.2-K V 2.1 proximity. Images are maximum intensity projections from Z-stacks spanning whole cells. Color coding as in ( C ). Right: quantification of PLA puncta density. n = 20 neurons per condition; two independent isolations. E Same experimental design as in ( D ), but assessing Ca V 2.1-K V 2.1 proximity. n = 20 (CTL), n = 20 (PFF), n = 21 (CTL;Rosco), and n = 20 (PFF;Rosco) neurons; two independent isolations. Error bars represent SEM. Statistical significance in panel ( A ) was determined using two-tailed Mann-Whitney test; panels ( C – E ) were analyzed using two-way ANOVA with appropriate post hoc tests. ns, not significant; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001. CTL, control; PFF, α-synuclein pre-formed fibril treatment; Rosco, roscovitine; pS603-K V 2.1, K V 2.1 phosphorylated at serine 603.

Article Snippet: For negative controls, Ca V 1.2 antibody (Alomone, ACC-003) was applied alone.

Techniques: Immunolabeling, Control, Incubation, Two Tailed Test, MANN-WHITNEY