Journal: Development (Cambridge, England)

Article Title: Neanderthal-derived variants increase SOX9 enhancer activity in craniofacial progenitors that shape jaw development

doi: 10.1242/dev.204779

Figure Lengend Snippet: EC1.45 Peak1-2 is active during embryonic facial development in the frontonasal region and adjacent to the developing jaw. (A) Schematic of the cross between the human EC1.45 Peak1-2 enhancer reporter Tg(HuEC1.45-P1P2:eGFP) and the Tg(sox10:mRFP) reporter. The Peak1-2 region is upstream of a minimal gata2 promoter driving eGFP expression and flanked by insulators and Tol2 sites. (B) Confocal microscopy images (maximum intensity projections) of the cranial region of embryos at 2, 3 and 4 dpf for Tg(HuEC1.45-P1P2:eGFP) crossed with Tg(sox10:mRFP) . Scale bars: 100 µm. (C) (i) Schematics of the zebrafish embryonic cranial region (ventral view) indicating the location of sox10:mRFP reporter activity in PCCs, which mature to cartilage templates from 2 to 4 dpf, and additional neural crest-derived mesenchymal populations (fn and o). See also . (ii) Human (Hs) embryonic lower jaw development at 6 weeks (upper; see also ) compared to adult jaw (lower, brown). (D) HCR RNA-FISH for eGFP and sox9a in zebrafish embryos at 2 dpf from the Tg(HuEC1.45-P1P2:eGFP) transgenic line. Top: 20× max projection images for the cranial region. Bottom: single z -plane images used for quantification, demonstrating overlap of EC1.45 activity with sox9a expression ( n =3 embryos). Scale bars: 100 µm for 20× images and 50 µm for 40× zoom. Arrow and outline highlight the region of eGFP and sox9a double-positive cells. (E) Quantification of the proportion of eGFP-positive cells from D that express only eGFP (4%) or are also positive for sox9a (96%), from 40× images ( n =3 embryos, data are mean±s.d.). b, bone; m, Meckel's cartilage; Md, mandibular prominence; p, palatoquadrate; e, ethmoid plate; c, ceratohyal; fn, frontonasal; o, oral; PA1, pharyngeal arch 1; BF, brightfield.

Article Snippet: Plasmid pCS2FA CO Tol2 TPase is available at Addgene ( plasmid #133032 ).

Techniques: Expressing, Confocal Microscopy, Activity Assay, Derivative Assay, Transgenic Assay

Journal: Current Research in Toxicology

Article Title: Application to developmental toxicity testing of a novel method for whole-brain imaging of microglia in zebrafish ☆

doi: 10.1016/j.crtox.2025.100276

Figure Lengend Snippet: Whole-brain imaging of microglia in live transgenic zebrafish at 5–6 dpf by two-photon microscopy. (A) Left panel: Structure of the transposon vector used to generate Tg (mpeg1:mVenus) zebrafish. Center and right panels: Representative 2D image of a Z-plane (center) and 3D image of microglia (right) in whole Tg (mpeg1:mVenus) brain. (B) Representative Z-plane image of Tg (eno2:Cerulean) brain. (C) Representative Z-plane (left) and 3D image (right) of SHG of Tg (mpeg1:mVenus) brain used for voxelation. (D) Schematic of the voxelation approach to regional mapping of microglia in the telencephalon (yellow), mesencephalon (cyan), diencephalon (magenta), and metencephalon (orange) in Tg (mpeg1:mVenus) (left panel) and the microglial morphology with the same color-code (right panel). (E) Representative images of microglial morphology (color-coded as in C) in the whole brain of zebrafish exposed to EtOH (291 mM for 8 h) or VPA (2 mM for 24 h). Ro = rostral; Ca = caudal; Lt = left; Rt = righ.t. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The Tol2 and mpeg1 genes were amplified from the Tol2-mpeg1-dendra2 plasmid ( ) (Addgene, Watertown, MA, USA) by inverse PCR using PrimeSTAR Max DNA Polymerase (Takara Bio, Shiga, Japan) and the primers mpeg1_infu_F1: 5′-GCG GCC GCG ACT CTA GAT-3′ and mpeg1_infu_R1: 5′-GGT GGC GGA TCC TTT TGC TG-3′.

Techniques: Imaging, Transgenic Assay, Microscopy, Plasmid Preparation