Journal: Cellular and Molecular Life Sciences

Article Title: Aβ1-16 controls synaptic vesicle pools at excitatory synapses via cholinergic modulation of synapsin phosphorylation

doi: 10.1007/s00018-021-03835-5

Figure Lengend Snippet: N-terminal portion of Aβ requires choline and nicotine to increase TRP and Ca 2+ influx via α7nAChRs. a Images show TRP labelled by Syt1 Ab loading (magenta) in rat cortical neurons treated either with vehicle or 200 pMAβ1-16 (1 h, 37 °C) in the neuronal growth media, in Tyrode´s buffer or in Tyrode´s buffer supplemented with 28 μM choline. Synapses were identified by co-staining for Syn1,2 (green). Scale bar 5 μm. b Quantification of the experiment described in a . c Ca 2+ imaging was done using Cal-520 AM indicator in HEK293T expressing α7nAChRs. Traces show the time course of the experiment. Arrows depict the sequential applications of human Aβ1-16, human/rat Aβ17-42, human Aβ1-42 (200 pM, 115 μl/s) or vehicle solution and nicotine (100 μM, 115 μl/s). d Quantification of nicotine-induced Cal-520 fluorescence in experiment shown in graph (C). Values in brackets refer to the number of analysed cells ( b) or recorded wells ( d ) obtained from two ( b ) or four (D) independent preparations. Boxes indicate the interquartile range with median, whiskers minimum and maximum values, and + shows the mean. Significance was assessed by one-way ANOVA followed by Dunnett´s post hoc test; marks (*** p < 0.001, ** p < 0.01) above boxes indicate comparisons to the Ctrl

Article Snippet: For immunocytochemistry and live staining, the following antibodies were used: rabbit primary antibodies against synaptotagmin 1 for uptake in mouse cells (Oyster550-labelled, 1:100, #105103C3, Synaptic Systems, Göttingen, Germany), synapsin 1,2 (1:1000, #106002, Synaptic Systems), phospho-synapsin 1 (pSyn1S553) (human pS553 corresponds to pS551 in rat; 1:1000, #ab32532, Abcam, Cambridge, UK), VGLUT1 (rat cultures, 1:1000, #135303, Synaptic Systems), VGAT (1:1000, #131003, Synaptic Systems).

Techniques: Staining, Imaging, Expressing, Fluorescence

Journal: Cellular and Molecular Life Sciences

Article Title: Aβ1-16 controls synaptic vesicle pools at excitatory synapses via cholinergic modulation of synapsin phosphorylation

doi: 10.1007/s00018-021-03835-5

Figure Lengend Snippet: Aβ-derived fragments differently affect phosphorylation of Syn1 at serine 551 and calcineurin activity. a Representative images of mature cortical neurons (17–22 DIV) treated with vehicle (Ctrl), rat Aβ1-40, rat Aβ1-42, human Aβ1-16, human/rat Aβ17-40, human/rat Aβ17-42 for 1 h at 37 °C and stained with antibodies recognizing Syn1 phosphorylation at serine 551 (pSyn1S551, green). Syn1,2 (magenta) was used as a synaptic marker. Scale bar 5 μm. b Quantification of the density and c IF intensity of synaptic puncta positive for pSyn1S551. (D) Quantification of calcineurin activity in cortical neurons (21DIV) treated with Aβ peptides, BgTx and PNU-120596. Data were normalized to the Ctrl. In plots, the boxes show the interquartile range and median, whiskers minimum and maximum values, and + indicates the mean. Values in brackets are the in b and c number of analysed cells obtained from three independent experiments and in d numbers of independent experiments. One-way ANOVA followed by Dunnett´s post hoc test was applied to assess statistical significance of comparison to Ctrl; * p < 0.05, ** p < 0.01, **** p < 0.0001

Article Snippet: For immunocytochemistry and live staining, the following antibodies were used: rabbit primary antibodies against synaptotagmin 1 for uptake in mouse cells (Oyster550-labelled, 1:100, #105103C3, Synaptic Systems, Göttingen, Germany), synapsin 1,2 (1:1000, #106002, Synaptic Systems), phospho-synapsin 1 (pSyn1S553) (human pS553 corresponds to pS551 in rat; 1:1000, #ab32532, Abcam, Cambridge, UK), VGLUT1 (rat cultures, 1:1000, #135303, Synaptic Systems), VGAT (1:1000, #131003, Synaptic Systems).

Techniques: Derivative Assay, Activity Assay, Staining, Marker

Journal: STAR Protocols

Article Title: Protocol for generating human cerebral organoids from two-dimensional cultures of pluripotent stem cells bypassing embryoid body aggregation

doi: 10.1016/j.xpro.2025.103678

Figure Lengend Snippet: Characterization of the main cell types in the hCOs at maturation (A) hCOs immunostained for cell cycle marker Ki67 (green) and neural precursor marker Sox2 (green), different neuronal markers (DCX, βIIITubulin and MAP2) (red) and synaptic marker (SYN1) (green). Cell nuclei were identified by Höechst 33258 staining (blue). VZs are marked with white dashed lines. Scale bar = 100 μm. (B) Whole-mount immunohistochemistry of hCOs stained for Sox2 (red) and MAP2 (green). Cell nuclei were identified by Höechst 33258 staining (blue). VZs are marked with white dashed lines. Scale bar = 100 μm. hCOs, human cerebral organoids.

Article Snippet: Rabbit polyclonal anti-Synapsin 1 (Syn1) (1:200) , Merck (Millipore) , Cat#AB1543; RRID: AB_2200400.

Techniques: Marker, Staining, Immunohistochemistry

Journal: STAR Protocols

Article Title: Protocol for generating human cerebral organoids from two-dimensional cultures of pluripotent stem cells bypassing embryoid body aggregation

doi: 10.1016/j.xpro.2025.103678

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-Synapsin 1 (Syn1) (1:200) , Merck (Millipore) , Cat#AB1543; RRID: AB_2200400.

Techniques: Recombinant, Knock-Out, Software

Journal: Cell

Article Title: Neuromodulator Signaling Bidirectionally Controls Vesicle Numbers in Human Synapses

doi: 10.1016/j.cell.2019.09.011

Figure Lengend Snippet: Highlights:

Article Snippet: Mouse monoclonal anti Synapsin-1 , Synaptic Systems , Clone 10.22.

Techniques: Transduction, Recombinant, Suspension, Plasmid Preparation, Expressing, Software

Journal: Cell Reports Medicine

Article Title: Human astrocytes and microglia show augmented ingestion of synapses in Alzheimer’s disease via MFG-E8

doi: 10.1016/j.xcrm.2023.101175

Figure Lengend Snippet: Astrocytes ingest more synapses in Alzheimer’s disease compared with midlife and aged controls (A) Confocal images of immunostaining with orthogonal views (left) and three-dimensional reconstructions of stacks (right) of midlife control, aged control, and Alzheimer’s disease showing ingestion of synapsin 1 (Syn1, green) by astrocytes (GFAP, magenta) in human brain sections. Aβ plaques and nuclei are counterstained with Thioflavin S and DAPI, respectively, shown in blue. Only ingested Syn1 is shown in 3D Imaris reconstructions. Representative images are from BA17. Scale bars represent 5 μm. (B) Quantification of synapsin 1 ingested by GFAP-positive astrocytes showed significantly increased levels in AD compared with both midlife controls (post hoc Tukey corrected tests after linear mixed effects model t = 8.138, p < 0.0001) and aged controls (t = 4.997, p < 0.0001), and also between midlife controls to aged controls (t = 3.68, p = 0.0015). (C) Statistically significant increase of synapsin 1 ingestion by astrocytes near Aβ plaques in AD cases (F[1,1202.02] = 13.6, p = 0.0002366). (D) The APOE4 genotype was associated with an increase in synapsin 1 colocalization inside GFAP-positive astrocytes (F[1,78.36] = 17.81, p = 6.5 × 10 −5 ). (E) BA17 (primary visual cortex) was associated with higher levels of synapsin 1 colocalization inside GFAP-positive astrocytes compared with BA20/21 (inferior temporal cortex) (F[1,2228.59] = 1382.4, p = 2.2 × 10 −16 ). Statistical comparisons were made using ANOVA after linear mixed effects model on Tukey transformed data with case as a random effect and disease, brain region, APOE4 status, gender, and age as fixed effects. Untransformed data are presented in graphs (B)–(E). All data are included in boxplots and case medians are shown in points. Data in this figure are pooled from both BA17 and BA20/21. Males are represented by circles and females by triangles. Biological replicates were human brain donors: n = 10 midlife controls, 19 aged controls, AD 31 cases.

Article Snippet: Rabbit anti Synapsin 1 (used 1:750) , Merck Millipore AB1543P , RRID: AB_90757.

Techniques: Immunostaining, Transformation Assay

Journal: Cell Reports Medicine

Article Title: Human astrocytes and microglia show augmented ingestion of synapses in Alzheimer’s disease via MFG-E8

doi: 10.1016/j.xcrm.2023.101175

Figure Lengend Snippet: Microglia ingest more synapses in Alzheimer’s disease compared with midlife and aged controls (A) Confocal images of immunostaining with orthogonal views (left) and three-dimensional reconstructions of stacks (right) of midlife control, aged control, and Alzheimer’s disease showing ingestion of synapsin 1 (Syn1, green) by microglia (CD68, magenta) in human brain sections. Aβ plaques and nuclei are counterstained with Thioflavin S and DAPI, respectively, shown in blue. Only ingested Syn1 is shown in 3D Imaris reconstructions. Representative images are from BA17. Scale bars represent 5 μm. (B) Quantification of synapsin 1 ingested by CD68-positive microglia showed significantly increased levels in AD compared with midlife (Tukey corrected post hoc test t = 3.56, p = 0.0024) and aged controls (t = 3.09, p = 0.0096). No statistical difference was seen between midlife controls and aged control (t = 0.93, p = 0.625). Data here are pooled from both BA17 and BA20/21, sample size corrected by mixed effected linear model. (C) Statistically significant increase of synapsin 1 ingestion by microglia near Aβ plaques in AD cases (ANOVA F[1,881.57] = 20.74, p = 6.01 × 10 −6 ). (D) The APOE4 genotype was associated with an increase in synapsin 1 colocalization inside CD68-positive microglia (F[1,42.86] = 5.84, p = 0.02). (E) BA17 (primary visual cortex) was associated with higher levels of synapsin 1 colocalization inside CD68-positive microglia compared with BA20/21 (inferior temporal cortex) (F[1,1973.76] = 67.82, p = 3.42 × 10 −16 ). Statistical comparisons were made using post hoc Tukey test or ANOVA after linear mixed effects model on Tukey transformed data with case as a random effect and disease, brain region, APOE4 status, gender, and age as fixed effects. Untransformed data are presented in graphs (B)–(E). Males are represented by circles and females by triangles. Biological replicates were human brain donors: n = 10 midlife controls, 17 aged controls, AD 22 cases.

Article Snippet: Rabbit anti Synapsin 1 (used 1:750) , Merck Millipore AB1543P , RRID: AB_90757.

Techniques: Immunostaining, Transformation Assay

Journal: Cell Reports Medicine

Article Title: Human astrocytes and microglia show augmented ingestion of synapses in Alzheimer’s disease via MFG-E8

doi: 10.1016/j.xcrm.2023.101175

Figure Lengend Snippet:

Article Snippet: Rabbit anti Synapsin 1 (used 1:750) , Merck Millipore AB1543P , RRID: AB_90757.

Techniques: Labeling, Blocking Assay, Cell Isolation, Recombinant, Plasmid Preparation, Protease Inhibitor, Western Blot, Molecular Weight, Marker, Software