SAS-1 loss leads to impaired centriole integrity during oogenesis.
<p><b>A.</b> Schematic of the gonad of a young adult hermaphrodite showing successive stages of gametogenesis, with early mitotic region, followed by early meiotic prophase, before loop region and cellularization. Mature oocytes traverse the spermatheca where fertilization takes pl...
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2025
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| Summary: | <p><b>A.</b> Schematic of the gonad of a young adult hermaphrodite showing successive stages of gametogenesis, with early mitotic region, followed by early meiotic prophase, before loop region and cellularization. Mature oocytes traverse the spermatheca where fertilization takes places. <b>B-D.</b> Representative immunofluorescence images of early mitotic region in fixed gonads of control (B), <i><i>sas-6(ok2554)</i></i> (C), and <i><i>sas-1(is13)</i></i> (D) animals expressing GFP::SAS-7, immunostained with antibodies against SAS-4 and GFP. Dotted lines highlight edges of gonad. Square insets on the right are ~ 1.5 times magnified, whereas dashed red circles exemplify regions measured for nuclear area reported in (E). Brightness and contrast adjusted individually for each image. <b>E.</b> Quantification of nuclear area in mitotic region of indicated genotypes. Number of nuclei: N<i> = </i>98 –control, from 5 gonads; 51 –<i><i>sas-6(ok2554)</i></i>, from 6 gonads; 101 –<i><i>sas-1(is13)</i></i>, from 5 gonads. Student’s two-tailed t-tests, whereby <i><i>P</i></i> < 0.001 (***). <b>F.</b> Number of nuclei in mitotic region of gonads of indicated genotypes. Number of gonads<i> = </i>9 –control and <i><i>sas-6(ok2554);</i></i> 10 –<i><i>sas-1(is13).</i></i> Student’s two-tailed t-tests, whereby <i><i>P</i></i> < 0.01 (**). The difference between control and <i><i>sas-1</i></i>(<i><i>is13</i></i>) is not significant (<i><i>P</i></i> = 0.68)<i>.</i> <b>G-I.</b> Representative U-Ex-STED images of centrioles in the mitotic region of control (G; N = 23) and <i><i>sas-1(is13)</i></i> mutants (H, I; N = 15), immunostained for SAS-4 and α/β-tubulin. Arrowheads: procentrioles; arrow: deformed and barely visible centriole. Note that U-Ex-STED might have caused fragile centrioles to further disintegrate as SAS-4::GFP levels in live imaging experiments are unchanged (see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1011912#pgen.1011912.s002" target="_blank">S2D</a>–<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1011912#pgen.1011912.s002" target="_blank">S2F Fig</a>). Scale bar is corrected for the expansion factor of 5.</p> |
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