Figure 2
- ID
- ZDB-FIG-230428-26
- Publication
- Yamaguchi et al., 2023 - Calaxin stabilizes the docking of outer arm dyneins onto ciliary doublet microtubule in vertebrates
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(A–C) Immunofluorescence microscopy of zebrafish spermatozoa. Scale bar: 20 μm. (A) Dnah8 was localized along the entire length of sperm flagella in WT. In calaxin-/-, Dnah8 was lost at the distal region of sperm flagella (white arrowheads). In arm4-/-, Dnah8 was lost (white asterisk). (B) Calaxin was localized along the entire length of sperm flagella in WT. In calaxin-/- and arm4-/-, Calaxin was lost (white asterisks). (C) Dnah2 was localized along the entire length of sperm flagella in WT, calaxin-/-, and arm4-/-. (D) Phase-contrast microscopy images of swimming spermatozoa (left column), traces of beating flagella (middle column), and shear angle plots of traced flagella (right column). Swimming spermatozoa were filmed using a high-speed camera at 1000 fps (frames per second). Shear angles were plotted against the distance from the flagellar base. In calaxin-/-, shear angle plots lost their slopes at the distal region of the flagella (black arrowhead). Scale bars: 10 μm. (E–G) Motilities of swimming spermatozoa were filmed using a high-speed camera at 200 fps and analyzed by CASA modified for zebrafish. For each zebrafish line, more than 600 spermatozoa were analyzed in total with 16 technical replicates. Spermatozoa with less than 20 μm/s velocities were considered immotile. (H) Ratio of motile spermatozoa. (I) Velocity of spermatozoa on averaged paths. The averaged paths were drawn by connecting the points of averaged sperm positions of contiguous 33 frames. (J) Frequencies at which sperm heads crossed their averaged paths. Boxes correspond to the first and third quartiles, lines inside the boxes indicate the medians, and whiskers extend to the full range of the data. p-Values were calculated with Tukey-Kramer test.
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