Journal: Science Advances

Article Title: Functional interrogation of neuronal connections by chemoptogenetic presynaptic ablation

doi: 10.1126/sciadv.aeb6755

Figure Lengend Snippet: ( A ) Schematic of the construct used to make the UAS:syp-dL5-mNeonGreen transgenic line. ( B ) Horizontal confocal section through the spinal cord of 3-dpf y417-Gal4, UAS:syp-dL5-mNeonGreen embryo (green), stained with anti-sv2 (magenta). Scale bar, 20 μm. Arrows show synaptic puncta from y417 neurons. “A” and “D” denote anterior-dorsal orientation. The boxed region is shown at a higher magnification in the right panel. Scale bar, 5 μm. ( C and D ) Maximum projection from the confocal z -stack of GFP and differential interference contrast channels in 4 dpf y252-Gal4, UAS:syp-dL5-mNeonGreen larvae: untreated control larva (C) and an MG-2I–treated larva exposed for 20 min to wide-field illumination at 3 dpf (D). Scale bar, 100 μm. A square ROI was cropped from each panel in (C) and (D) and is shown at a higher magnification in (Ca) and (Da). ( E ) SLC [ n = 25 and 25 for ctl (dL5 − /Mg2I + ) and abl (dL5 + /Mg2I + )] and LLC ( n = 25 and 20) escape responsiveness and PPI ( n = 14 and 16) in control and wide-field light–exposed y252-Gal4, UAS:syp-dL5-mNeonGreen larvae. MWU, significant P value indicated and * P < 0.001. ( F ) Maximum projection of 80-μm confocal stacks from 4-dpf tph2:Gal4, UAS:syp-dL5, UAS:Kaede Red untreated larvae and in larvae exposed to NIR illumination in the indicated region of the left hindbrain at 3 dpf. Scale bars, 100 μm. Ra, raphe neurons. ( G ) Number of neurons in dorsal raphe 24 hours after photoablation of neuropil in the region indicated in (F) in untreated [ctl (dL5 − /Mg2I + ), n = 9] and photoablated larvae [abl (dL5 + /Mg2I + ), n = 8]. Welch t test, P = 0.50.

Article Snippet: Primary antibody incubation with a mouse anti–SV2 antibody (1:200, Developmental Studies Hybridoma Bank, Iowa City, IA) in PBS-T (PBS and 1% Triton) for 2 days at 4°C.

Techniques: Construct, Transgenic Assay, Staining, Control

Journal: Cell Reports

Article Title: The insertion of an ATTTC repeat in an Alu element hyperactivates a neurodevelopmental enhancer in spinocerebellar ataxia type 37

doi: 10.1016/j.celrep.2026.117146

Figure Lengend Snippet: DAB1 overexpression induces axonal defects during development (A) Schematic representation of cerebellar and cerebellar-like structures in zebrafish. Granule cell (GC) axons, named parallel fibers (PFs), innervate Purkinje cells (PCs) in the molecular cell layer. Axons from GCs in eminentia granularis (EG) innervate a type of Purkinje-like neuron called crest cells (CrCs). (B) Representative confocal images of zebrafish larvae, from a GFP + cerebellar granule cell line, microinjected with control or human DAB1 mRNA at 5 dpf; axonal tracts of GCs from EG are indicated with orange (absent) or gray (present) arrows. (C) Percentage of present or absent GC axonal tracts that innervate PC-like neurons (graphic representation of data by mean ± SD of at least 20 embryos/condition/replicate; three independent experiments; ꭕ 2 test for the percentage of GC axonal tracts, present or absent; ∗∗∗ p < 0.001). (D) Schematics of muscle innervation by PMN axon tracts in zebrafish larvae at 24 hpf. (E) Representative confocal images (40× objective) of PMN axon tracts from the 6-somites-spanning region anterior to the cloaca from control or DAB1 mRNA-injected embryos; whole-mounted immunofluorescence with SV2 antibody, representative z-projection images (maximum intensity). (F) Length of zebrafish PMN axon tracts in control and DAB1 mRNA-microinjected embryos ( n = 23 embryos for control and n = 24 embryos in DAB1 mRNA condition; four independent experiments; independent t test, ∗ p < 0.05; data are represented by the mean ± SD). (G) Proposed mechanistic model showing disruption of DRL function by the (ATTTC) n , which hyperactivates the neurodevelopmental enhancer, leading to DAB1 upregulation and DAB1 protein accumulation in axons, likely contributing to SCA37. See also .

Article Snippet: Embryos were incubated with anti-synaptic vesicle 2 (SV2) antibody (1:200, #AB2315387, Developmental Studies Hybridoma Bank) diluted in blocking solution overnight, at 4°C.

Techniques: Over Expression, Control, Injection, Immunofluorescence, Disruption

Journal: Cell Reports

Article Title: The insertion of an ATTTC repeat in an Alu element hyperactivates a neurodevelopmental enhancer in spinocerebellar ataxia type 37

doi: 10.1016/j.celrep.2026.117146

Figure Lengend Snippet: DAB1 overexpression induces axonal defects during development (A) Schematic representation of cerebellar and cerebellar-like structures in zebrafish. Granule cell (GC) axons, named parallel fibers (PFs), innervate Purkinje cells (PCs) in the molecular cell layer. Axons from GCs in eminentia granularis (EG) innervate a type of Purkinje-like neuron called crest cells (CrCs). (B) Representative confocal images of zebrafish larvae, from a GFP + cerebellar granule cell line, microinjected with control or human DAB1 mRNA at 5 dpf; axonal tracts of GCs from EG are indicated with orange (absent) or gray (present) arrows. (C) Percentage of present or absent GC axonal tracts that innervate PC-like neurons (graphic representation of data by mean ± SD of at least 20 embryos/condition/replicate; three independent experiments; ꭕ 2 test for the percentage of GC axonal tracts, present or absent; ∗∗∗ p < 0.001). (D) Schematics of muscle innervation by PMN axon tracts in zebrafish larvae at 24 hpf. (E) Representative confocal images (40× objective) of PMN axon tracts from the 6-somites-spanning region anterior to the cloaca from control or DAB1 mRNA-injected embryos; whole-mounted immunofluorescence with SV2 antibody, representative z-projection images (maximum intensity). (F) Length of zebrafish PMN axon tracts in control and DAB1 mRNA-microinjected embryos ( n = 23 embryos for control and n = 24 embryos in DAB1 mRNA condition; four independent experiments; independent t test, ∗ p < 0.05; data are represented by the mean ± SD). (G) Proposed mechanistic model showing disruption of DRL function by the (ATTTC) n , which hyperactivates the neurodevelopmental enhancer, leading to DAB1 upregulation and DAB1 protein accumulation in axons, likely contributing to SCA37. See also .

Article Snippet: Anti-Synaptic Vesicle 2 (SV2) primary antibody , Developmental Studies Hybridoma Bank , Cat #SV2;RRID:AB_2315387.

Techniques: Over Expression, Control, Injection, Immunofluorescence, Disruption

Journal: PLOS One

Article Title: Dysregulated lactate metabolism synergizes with ALS genetic risk factors to accelerate motor decline

doi: 10.1371/journal.pone.0347135

Figure Lengend Snippet: Motor function (latency to fall from an inverted screen) measured in 3-month-old WT, TDP43 Q331K/+ , LDHB MNKO and TDP43 Q331K/+ ; LDHB MNKO mice (A), electrophysiological recording performed on 3-month-old animals to determine the difference in CMAP at the ankle and sciatic notch ( B & C ) and Sensory Nerve Action Potential (SNAP) at the tail (n = 10-12) (D). NMJs on lumbrical muscles stained with anti-NF, anti-SV2 and BTX to compare innervation of 3-month-old mice of different genotypes (n = 3; statistical comparison is between fully innervated endplates). Statistical significance was determined by pairwise Fisher’s exact tests on raw counts, with a Bonferroni correction applied for multiple comparisons. (E-I). Spinal cords stained to detect TDP43 and nuclei (DAPI) indicate no mislocalization in 6-month-old LDHB MNKO; TDP43 Q331K/+ or LDHB MNKO mice (J & K) . Representative images of 40x and 63x toluidine blue-stained sections of tibial nerves from 3-month-old animals (L-O) . Average g-ratio indicates no difference in degeneration between genotypes (n = 3). Unless otherwise mentioned, all statistical significance was determined by one-way ANOVA with Tukey’s multiple comparisons test (P) . ***p < 0.001, **p < 0.01.

Article Snippet: Following three rinses with Phosphate buffered saline (PBS), lumbrical muscles were dissected for staining with anti-SV2 (Developmental Studies Hybridoma Bank AB2315387, 1:200), anti-2H3 (Developmental Studies Hybridoma Bank AB2314897, 1:100) and α-bungarotoxin (BTX) (Biotium 00006, 1:500) as described previously [ ].

Techniques: Muscles, Staining, Comparison