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

Journal: Frontiers in Physiology

Article Title: Transcriptomic profiling of the sex-linked biological pathways of severe pulmonary arterial hypertension associated with endothelial cell caveolin-1 depletion and chronic hypoxia

doi: 10.3389/fphys.2026.1794886

Figure Lengend Snippet: Effect of Caveolin-1 (Cav1) knockout and reconstitution on mouse survival in hypoxia. Representative Western Blots against CAV1 in (A) male (n = 3 – 9) and (B) female (n=4-7) mice after 6 weeks of hypoxia. All western blot images used for quantification are available as Source Data. Data is quantified relative to whole protein using ponceau staining. Post-hoc 2-way analysis of variance (ANOVA) with Bonferonni’s multiple comparisons test where *P-value < 0.05, ****P-value < 0.0001. Data presented as mean ± standard error of the mean (SEM). (C) Survival curves of wildtype (n=30), Caveolin-1-knockout (n=37), and reconstituted mice (n=12) in hypoxia (10% oxygen) over 60 days. (D) Survival curve comparing male (n=16) and female Caveolin-1 (n=21) knockout over 60 days of hypoxia. Post-hoc survival curve analysis was performed with Mantel-Cox comparisons.

Article Snippet: Cav1 -/- (Cav1 tm1Mls /J) and B6/129SJ2 wild-type control mice were originally purchased from Jackson Labs. Cav1-RC mice ( ) were obtained from William Sessa, Yale University.

Techniques: Knock-Out, Western Blot, Staining

Journal: Frontiers in Physiology

Article Title: Transcriptomic profiling of the sex-linked biological pathways of severe pulmonary arterial hypertension associated with endothelial cell caveolin-1 depletion and chronic hypoxia

doi: 10.3389/fphys.2026.1794886

Figure Lengend Snippet: Right ventricular (RV) function and structure of Caveolin-1 (Cav1) global knockout or Caveolin-1 endothelial-specific reconstituted male and female mice exposed to 6 weeks of hypoxia. Control (Con), Cav1 global knockout (KO), and Caveolin-1 endothelial-specific reconstituted (RC) are plotted by normoxia (Normoxia), white bars, or hypoxia (Hypo), dotted bars. (A) Right ventricular systolic pressure (RVSP) (n = 4 – 16) (B) Right ventricular free wall thickness (RVFWT) (n = 4 – 11) (C) Right Ventricular Hypertrophy (RVH) as determined by the weight ratio of the right ventricle divided by the sum of left ventricle and septum (RV/(LV+S)) (n = 4 – 25) (D) Pulmonary acceleration time (PAT) (n = 4 – 15) (E) Tricuspid annular plane systolic excursion (TAPSE) (n = 4 – 15). Post-hoc 3-way ANOVA analysis with Bonferonni’s multiple comparisons tests where *P-value < 0.05, **P-value < 0.01, ***P-value <0.001, ****P-value < 0.0001. Data presented as mean ± standard error of the mean (SEM).

Article Snippet: Cav1 -/- (Cav1 tm1Mls /J) and B6/129SJ2 wild-type control mice were originally purchased from Jackson Labs. Cav1-RC mice ( ) were obtained from William Sessa, Yale University.

Techniques: Knock-Out, Control

Journal: Frontiers in Physiology

Article Title: Transcriptomic profiling of the sex-linked biological pathways of severe pulmonary arterial hypertension associated with endothelial cell caveolin-1 depletion and chronic hypoxia

doi: 10.3389/fphys.2026.1794886

Figure Lengend Snippet: Cardiac echocardiography of Caveolin-1 (Cav1) global knockout or Caveolin-1 endothelial-specific reconstituted male and female mice exposed to 6 weeks of hypoxia. Control (Con), Cav1 global knockout (KO), and Caveolin-1 endothelial-specific reconstituted (RC) are plotted by normoxia (Normoxia), white bars, or hypoxia (Hypo), dotted bars. (A) Cardiac Output (CO) (n = 4 – 15) (B) Ejection Fraction (EF) (n = 4 – 15) C) Fractional Shortening (FS) (n = 4 – 15) (D) Stroke Volume (SV) (n = 4 – 15) (E) Isovolumetric relaxation time (IVRT) (n = 4 – 25). Post-hoc 3-way ANOVA analysis with Bonferonni’s multiple comparisons test where *P-value < 0.05, **P-value < 0.01, ***P-value < 0.001, ****P-value < 0.0001. Data presented as mean ± standard error of the mean (SEM).

Article Snippet: Cav1 -/- (Cav1 tm1Mls /J) and B6/129SJ2 wild-type control mice were originally purchased from Jackson Labs. Cav1-RC mice ( ) were obtained from William Sessa, Yale University.

Techniques: Knock-Out, Control

Journal: Frontiers in Physiology

Article Title: Transcriptomic profiling of the sex-linked biological pathways of severe pulmonary arterial hypertension associated with endothelial cell caveolin-1 depletion and chronic hypoxia

doi: 10.3389/fphys.2026.1794886

Figure Lengend Snippet: Blood gas measurements of Caveolin-1 (Cav1) global knockout or Caveolin-1 endothelial-specific reconstituted male and female mice exposed to 6 weeks of hypoxia. Control (Con), Cav1 global knockout (KO), and Caveolin-1 endothelial-specific reconstituted (RC) are plotted by normoxia (Normoxia, white bars), or hypoxia (Hypo, dotted bars). (A) Oxygen saturation (sO2) (n = 3 – 8) (B) Partial pressure of oxygen (pO2) (n = 4 – 7) (C) Partial pressure of carbon dioxide (pCO2) (n = 3 – 8) (D) pH levels (pH) (n = 4 – 8) (E) Extracellular base excess (BeCef) (n = 3 – 8) (F) Bicarbonate (HCO3) (n = 3 – 8) (G) Lactate (n = 3 – 7) (H) Hemoglobin (Hgb) (n = 5 – 9) (I) Hemocrit (Hct) (n = 5 – 9). Post-hoc 3-way ANOVA analysis with Bonferonni’s multiple comparisons test where *P-value < 0.05, **P-value < 0.01, ****P-value < 0.0001. Data presented as mean ± standard error of the mean SEM).

Article Snippet: Cav1 -/- (Cav1 tm1Mls /J) and B6/129SJ2 wild-type control mice were originally purchased from Jackson Labs. Cav1-RC mice ( ) were obtained from William Sessa, Yale University.

Techniques: Knock-Out, Control

Journal: Frontiers in Physiology

Article Title: Transcriptomic profiling of the sex-linked biological pathways of severe pulmonary arterial hypertension associated with endothelial cell caveolin-1 depletion and chronic hypoxia

doi: 10.3389/fphys.2026.1794886

Figure Lengend Snippet: Pairwise comparison of differentially expressed genes between Caveolin-1 knockout (Cav1-KO) and wildtype control (Control) mouse lungs in normoxic conditions. Male (control n=3, Cav1-KO n=3) and female (control n=3, Cav1-KO n=2) comparisons were performed separately. The top 50 differentially expressed genes (DEG) upregulated and downregulated are shown in descending order of significance based on adjusted P-value (P adj ) for (A) female and (B) male lungs. DEG thresholds were set at Log2Foldchange > 1, P adj <0.05. Z-score coloring represents relative gene expression across all groups. Samples are sorted by complete hierarchical clustering. Gene Ontology (GO) analysis for (C) Female and (D) Male samples are shown as the top 20 gene ontologies in descending order of P adj . The X-axis represents the number of DEGs in this comparison that underlies the significant GO.

Article Snippet: Cav1 -/- (Cav1 tm1Mls /J) and B6/129SJ2 wild-type control mice were originally purchased from Jackson Labs. Cav1-RC mice ( ) were obtained from William Sessa, Yale University.

Techniques: Comparison, Knock-Out, Control, Gene Expression

Journal: Frontiers in Physiology

Article Title: Transcriptomic profiling of the sex-linked biological pathways of severe pulmonary arterial hypertension associated with endothelial cell caveolin-1 depletion and chronic hypoxia

doi: 10.3389/fphys.2026.1794886

Figure Lengend Snippet: Pairwise comparison of differentially expressed genes between Caveolin-1 knockout (Cav1-KO) and wildtype control (Control) mouse lungs in hypoxic conditions. Male and female comparisons were performed separately (n=3 in each group). The top 50 differentially expressed genes (DEG) upregulated and downregulated are shown in descending order of significance based on adjusted P-value (P adj ) for (A) female and (B) male lungs. DEG thresholds were set at Log2Foldchange > 1, P adj <0.05. Z-score coloring represents relative gene expression across all groups. Samples are sorted by complete hierarchical clustering. Gene Ontology (GO) analysis for (C) Female and (D) Male samples are shown as the top 20 gene ontologies in descending order of P adj . The X-axis represents the number of DEGs in this comparison that underlies the significant GO.

Article Snippet: Cav1 -/- (Cav1 tm1Mls /J) and B6/129SJ2 wild-type control mice were originally purchased from Jackson Labs. Cav1-RC mice ( ) were obtained from William Sessa, Yale University.

Techniques: Comparison, Knock-Out, Control, Gene Expression

Journal: Frontiers in Physiology

Article Title: Transcriptomic profiling of the sex-linked biological pathways of severe pulmonary arterial hypertension associated with endothelial cell caveolin-1 depletion and chronic hypoxia

doi: 10.3389/fphys.2026.1794886

Figure Lengend Snippet: Pairwise comparison of differentially expressed genes between endothelial Caveolin-1 reconsituted (Cav1-RC) and Caveolin-1 knockout (Cav1-KO) mouse lungs in normoxic conditions. Male (Cav1-KO n=3, Cav1-RC n=3) and female (Cav1-KO n=2, Cav1-RC n=3) comparisons were performed separately. The top 50 differentially expressed genes (DEG) upregulated and downregulated are shown in descending order of significance based on adjusted P-value (P adj ) for (A) female and (B) male lungs. DEG thresholds were set at Log2Foldchange > 1, P adj <0.05. Z-score coloring represents relative gene expression across all groups. Samples are sorted by complete hierarchical clustering. Gene Ontology (GO) analysis for (C) Female and (D) Male samples are shown as the top 20 gene ontologies in descending order of P adj . The X-axis represents the number of DEGs in this comparison that underlies the significant GO.

Article Snippet: Cav1 -/- (Cav1 tm1Mls /J) and B6/129SJ2 wild-type control mice were originally purchased from Jackson Labs. Cav1-RC mice ( ) were obtained from William Sessa, Yale University.

Techniques: Comparison, Knock-Out, Gene Expression

Journal: Frontiers in Physiology

Article Title: Transcriptomic profiling of the sex-linked biological pathways of severe pulmonary arterial hypertension associated with endothelial cell caveolin-1 depletion and chronic hypoxia

doi: 10.3389/fphys.2026.1794886

Figure Lengend Snippet: Pairwise comparison of differentially expressed genes endothelial Caveolin-1 reconsituted (Cav1-RC) and Caveolin-1 knockout (Cav1-KO) mouse lungs in hypoxic conditions. Male and female comparisons were performed separately (n=3 for each group). The top 50 differentially expressed genes (DEG) upregulated and downregulated are shown in descending order of significance based on adjusted P-value (P adj ) for (A) female and (B) male lungs. DEG thresholds were set at Log2Foldchange > 1, P adj <0.05. Z-score coloring represents relative gene expression across all groups. Samples are sorted by complete hierarchical clustering. Gene Ontology (GO) analysis for (C) Female and (D) Male samples are shown as the top 20 gene ontologies in descending order of P adj . The X-axis represents the number of DEGs in this comparison that underlies the significant GO.

Article Snippet: Cav1 -/- (Cav1 tm1Mls /J) and B6/129SJ2 wild-type control mice were originally purchased from Jackson Labs. Cav1-RC mice ( ) were obtained from William Sessa, Yale University.

Techniques: Comparison, Knock-Out, Gene Expression

Journal: Frontiers in Physiology

Article Title: Transcriptomic profiling of the sex-linked biological pathways of severe pulmonary arterial hypertension associated with endothelial cell caveolin-1 depletion and chronic hypoxia

doi: 10.3389/fphys.2026.1794886

Figure Lengend Snippet: Summary of the effect of Caveolin-1 (Cav1) genetic status, hypoxia, and sex on the expression of known pulmonary arterial hypertension (PAH) risk factors and summary figure of transcriptional changes found in Cav1 genetic modulation. (A) A heatmap of genes associated with heritable PAH risk. Each group represents the average expression of these genes across all samples in the group. Genes are organized by hierarchical clustering to demonstrate similar patterns of expression across the groups. (n = 2 – 3) (B) Summary figure showing which transcriptional pathways are modulated during global Cav1 knockout or endothelial Cav1 reconstitution. Gene ontologies are distinguished by which sex, or both, they show significant change.

Article Snippet: Cav1 -/- (Cav1 tm1Mls /J) and B6/129SJ2 wild-type control mice were originally purchased from Jackson Labs. Cav1-RC mice ( ) were obtained from William Sessa, Yale University.

Techniques: Expressing, Knock-Out