Journal: bioRxiv

Article Title: Development of the zebrafish anterior lateral line system is influenced by underlying cranial neural crest

doi: 10.1101/2025.02.11.637483

Figure Lengend Snippet: ( A-E ) Confocal microscopy time-lapse images of the cranial region of a Tg(cldnB:GFP;dRA:GFP;cldnB:H2A-mCherry) triple transgenic specimen, in lateral view with anterior to the left. Maximum projections are shown at 25 hpf (A), 28.7 hpf (B), 33 hpf (C), 36.7 hpf (D), and 42 hpf (E). A’-E’ are single channel images of the lateral line ( cldnB:GFP and dRA:GFP ) alone. Neuromasts are indicated with white arrowheads and labeled (SO: supraorbital neuromast; IO: infraorbital neuromast; O: otic neuromast). The supraorbital and infraorbital primordia are indicated (SOp, IOp). ( F-I ) Single Plane Illumination Microscopy (SPIM) time-lapse images of the cranial region of a Tg(sox10:NTRmCherry;cldnB:GFP) double transgenic specimen, in lateral view with anterior to the left, allowing comparison of the development of the lateral line system ( cldnB:GFP ; yellow) and neural crest ( sox10:NTRmCh ; magenta) over developmental time. Maximum projections (imaged using the 20x objective) are shown at 33 hpf (F), 37 hpf (G), 40.7 hpf (H), and 46.3 hpf (I). F’-I’ are single channel images of the lateral line ( cldnB:GFP ) alone. Neuromasts are indicated with white arrowheads and labeled. Small white arrowheads in F’ and G’ indicate the elongating ridge of the infraorbital line, which can be seen condensing to form the IO2 neuromast in G’-I’. Note budding of the SO2 neuromast in G’ and H’, extending a primordium (SO1p) that then forms the SO1 neuromast, I’. Abbreviations are as follows:-e: lens of the eye; ov: otic vesicle; ob: olfactory bulb. Scale bars are 50 µm.

Article Snippet: Images were captured with either a Zeiss Lightsheet Z.1 (with 20x objective) or a Zeiss Lightsheet 7 (with 10x objective) single-plane illumination microscope, each equipped with tandem PCO.edge sCMOS cameras (PCO.Imaging, Kelheim, Germany).

Techniques: Confocal Microscopy, Transgenic Assay, Labeling, Microscopy, Comparison

Journal: bioRxiv

Article Title: Development of the zebrafish anterior lateral line system is influenced by underlying cranial neural crest

doi: 10.1101/2025.02.11.637483

Figure Lengend Snippet: ( A ) Confocal maximum projection (40x objective), in dorsal view, of the left hand side of the anterior cranial region of a 6 mm SL Tg(sox10Cre;dsRed/EGFP ) specimen labeled with the vital dye TMRE, a mitochondrial marker labeling neuromasts. Canal neuromasts SO1 and SO2 are shown (TMRE, yellow) alongside neural crest-derived tissue ( sox10Cre;EGFP , magenta). Because TMRE is highly fluorescent in the rhodamine channel (546 nm), the sox10Cre:dsRed signal present throughout the specimen is not visible with the imaging parameters selected. Ob = olfactory bulb. ( B ) Large format lightsheet (4x objective), maximum projection of the right hand side of the cranium (boxed area in C) of a 22 mm SL adult Tg(sox10Cre;dsRed/EGFP ) specimen (of n=5). Neural crest cell-derived tissue is labeled with sox10Cre;EGFP (magenta), neuromasts are labeled with anti-Otoferlin antibody (yellow), in dorsal view with anterior to the left. ( B’ ) EGFP channel showing neural crest cell-derived tissue. ( B’’ ) Otoferlin channel showing neuromasts (in white dashed-line boxes). ( C ) Schematic of the supraorbital canals and canal neuromasts (yellow) in an adult zebrafish. The large frontal and small anterior nasal bones (next to the nostrils, nos) are indicated. Cell-type contributions to the bones are as indicated, neural crest in magenta, mesoderm in blue. ( D-G’ ) Confocal maximum projection (40x objective), in dorsal view with anterior to the left, of each of the canal neuromasts shown in B’’ (boxed regions). The white dashed lines in D-G indicate the supraorbital canal; D’-G’ show the sox10Cre;EGF P channel alone. Orthogonal (yz) sections, taken adjacent to each neuromast (at level of yellow arrows), are provided to the right of each panel. Scale bars are 50 µm for panels A, D-G’’ and 200 µm for panels B-B’’.

Article Snippet: Images were captured with either a Zeiss Lightsheet Z.1 (with 20x objective) or a Zeiss Lightsheet 7 (with 10x objective) single-plane illumination microscope, each equipped with tandem PCO.edge sCMOS cameras (PCO.Imaging, Kelheim, Germany).

Techniques: Labeling, Marker, Derivative Assay, Imaging