Journal: bioRxiv

Article Title: Wnt/β-catenin signaling promotes zebrafish osteoblast dedifferentiation by wnt10a -mediated inhibition of NF-κB

doi: 10.64898/2025.12.29.696582

Figure Lengend Snippet: A) RNASeq (left) and RT-qPCR (right) reveal upregulation of the NF-κB signaling target genes nfkbiaa and nfkbiab in osteoblasts sorted from bglap:GFP fish heterozygous for the wnt10a mutation relative to their wild-type siblings at 1 dpa. nE (qPCR biological replicates) = 3, nA = 10 per replicate. ΔΔCt values are normalized to the mean of the wildtype at 1 dpa. Error bars, Mean ± SEM. Two tailed Student’s t-test. B) Osteoblast dedifferentiation, as measured by bglap downregulation in segment -1 revealed by HCR in situ hybridization, is inhibited in fish treated with the Wnt inhibitor IWR-1, while IP injection of the NF-κB inhibitor Bay-11 slightly but significantly enhances dedifferentiation. Treatment with both inhibitors yields results similar to those of Bay-11 alone. nE = 3 (except for 2 for IWR-1), nA = 18 total per group (12 for IWR-1), nR = 33 (DMSO), 24 (IWR-1), 37 (Bay-11), 33 (both). C) Overexpression of wnt10a using hs:wnt10a fish is sufficient to cause downregulation of bglapl detected by HCR in situ hybridization in non-injured fins, relative to heat-shocked wild-type fish. HCR signals were quantified in a bony segment (“B”) that is located at the same proximal-distal position as “segment -1” in amputated fins. nE = 2, nA = 12 total per group, nR = 22 total per group. Dashed line, joints. Scale bar, 100 µm. D) Immunofluorescence on cryosections of hs:wnt10a transgenic hearts reveals increased embryonic myosin heavy chain (embMHC) expression in Myl7+ cardiomyocytes at the wound border at 7 days post injury (dpi). Plots show the ventricular area covered by anti-embMHC staining relative to the 150 µm wound border zone area occupied by Myl7+ myocardium. nE = 2, nA = 13 wild-type, 11 hs:wnt10a . Scale bar, 100 µm. (B, C, D) Error bars, mean ± 95% CI. Two tailed Student’s t-test.

Article Snippet: Primary antibodies against embMHC Mouse monoclonal (MYH7 DSHB, N2.261, RRID:AB_531790), and Myl7 Rabbit polyclonal (GeneTex, GTX128346, RRID:AB_2885759) were diluted in PEMTx/normal goad serum and applied overnight at 4 °C.

Techniques: Quantitative RT-PCR, Mutagenesis, Two Tailed Test, In Situ Hybridization, Injection, Over Expression, Immunofluorescence, Transgenic Assay, Expressing, Staining

Journal: bioRxiv

Article Title: Head stabilization behavior and underlying circuit mechanisms in larval zebrafish

doi: 10.1101/2025.11.12.688155

Figure Lengend Snippet: (A) Immunohistochemistry images (confocal stacked images) of Tg( smyhc2 :loxP-RFP-loxP-DTA) fish stained with S58 antibody. Images show fish without Cre (control, top row), with tbx2b :Cre (PHM-ablated, middle row), and with zic1 :Cre (SCA-ablated, bottom row). Left three columns show lateral-view images, with rostral to the left and dorsal to the top. Right three columns show dorsal-view images, with rostral to the left. Scale bars, 200 μm. (B) Body bend angle traces of control (top), PHM-ablated (middle), and SCA-ablated fish (bottom) in response to pitch tilts under head-embedded conditions. For each group, trials from the same fish are shown. Four to five trials are shown for each condition (six conditions total). (C) Pairwise comparison of maximum amplitudes of ventral and dorsal flexion angles between control and muscle-ablated animals. Mean values of three to five trials are shown for each condition. Non-ablated (Cre-negative) siblings were used as controls. Orange: PHM-ablated fish (9 fish), green: SCA-ablated fish (11 fish), black: control groups (9 fish for each group). For ventral flexion: p = 0.001 (PHM ablation), p = 0.37 (SCA ablation); for dorsal flexion: p = 0.60 (PHM ablation), p = 0.0003 (SCA ablation) (Wilcoxon exact rank-sum test). (D) Summary of muscles involved in the body flexions. The ventral flexion during head-up tilts is produced by PHMs, whereas the dorsal flexion during head-down tilts is produced by SCAs.

Article Snippet: The S58 monoclonal antibody supernatant (primary antibody; DSHB, RRID: AB_528377), which labels slow muscle fibers , was used at a 1:10 dilution.

Techniques: Immunohistochemistry, Staining, Control, Comparison, Muscles, Produced