Figure 1.
Loss of the rhoGEF rhgf-1 rescues spermatheca functions in the RhoGAP spv-1 loss of function mutant. (A) The reproductive system of an adult hermaphrodite C. elegans outlined on stitched DIC microscopy images. Scale bar, 50 µm. (B) Representative stitched DIC images of embryos from wild-type and spv-1(ok1498) loss-of-function worms treated with control (empty vector) or rhgf-1(RNAi). Black arrows point to abnormally shaped embryos. Scale bar, 50 µm. (C) Embryonic lethality of the progeny of wild-type or rhgf-1(ok880) loss-of-function worms treated with control or spv-1(RNAi). Nrepeats ≥ 5, Nmothers ≥ 150, Nembryos ≥ 990. Error bars are ± SEM. Statistical comparisons were performed by Brown-Forsythe ANOVA-Dunnett’s T3 multiple comparisons test. (D) Representative time points from DIC movies of the ovulation process in wild type, spv-1(1498), rhgf-1(ok880), and the double mutant spv-1(1498);rhgf-1(ok880). Time zero (0 s) is the time when the distal valve closes. The spermatheca is pseudo colored pink and the ovulating oocyte/embryo is pseudo colored green. Special phenotypes are labeled. Distal and proximal parts of the spermatheca are labeled. Lines labeled W1 and W2 relate to the measurement of constriction magnitude. Scale bar, 10 µm. Full movies are in Video 1. (E) Dwell time in wild-type, spv-1(1498), rhgf-1(ok880), and spv-1(1498);rhgf-1(ok880) day 1 adults, measured as the duration the oocyte resides in the spermatheca, from distal valve closure until the opening of the sp-ut valve. N ≥ 13. Error bars are mean ± SEM. Statistical comparisons were performed by One-way ANOVA-Tukey’s multiple comparisons test. (F) Constriction magnitude in wild type, spv-1(1498), rhgf-1(ok880), and spv-1(1498);rhgf-1(ok880), measured as the ratio of the width of the distal quarter of the spermatheca (W1) to the width of the proximal quarter of the spermatheca (W2) before the embryo starts to transit out. N ≥ 13. Error bars are mean ± SEM. Statistical comparisons were performed by Brown-Forsythe ANOVA-Dunnett’s T3 multiple comparisons test. (G) Successful embryos transit through the spermatheca of wild-type, spv-1(1498), rhgf-1(ok880), and spv-1(1498);rhgf-1(ok880) worms. N ≥ 14. Statistical comparisons were performed by individuals’ chi-square tests. Stars designate statistical significance (**** P < 0.0001, *** P < 0.001, ** P < 0.01, ns P > 0.05). N, number of samples analyzed.

Loss of the rhoGEF rhgf-1 rescues spermatheca functions in the RhoGAP spv-1 loss of function mutant. (A) The reproductive system of an adult hermaphrodite C. elegans outlined on stitched DIC microscopy images. Scale bar, 50 µm. (B) Representative stitched DIC images of embryos from wild-type and spv-1(ok1498) loss-of-function worms treated with control (empty vector) or rhgf-1(RNAi). Black arrows point to abnormally shaped embryos. Scale bar, 50 µm. (C) Embryonic lethality of the progeny of wild-type or rhgf-1(ok880) loss-of-function worms treated with control or spv-1(RNAi). Nrepeats ≥ 5, Nmothers ≥ 150, Nembryos ≥ 990. Error bars are ± SEM. Statistical comparisons were performed by Brown-Forsythe ANOVA-Dunnett’s T3 multiple comparisons test. (D) Representative time points from DIC movies of the ovulation process in wild type, spv-1(1498), rhgf-1(ok880), and the double mutant spv-1(1498);rhgf-1(ok880). Time zero (0 s) is the time when the distal valve closes. The spermatheca is pseudo colored pink and the ovulating oocyte/embryo is pseudo colored green. Special phenotypes are labeled. Distal and proximal parts of the spermatheca are labeled. Lines labeled W1 and W2 relate to the measurement of constriction magnitude. Scale bar, 10 µm. Full movies are in Video 1. (E) Dwell time in wild-type, spv-1(1498), rhgf-1(ok880), and spv-1(1498);rhgf-1(ok880) day 1 adults, measured as the duration the oocyte resides in the spermatheca, from distal valve closure until the opening of the sp-ut valve. N ≥ 13. Error bars are mean ± SEM. Statistical comparisons were performed by One-way ANOVA-Tukey’s multiple comparisons test. (F) Constriction magnitude in wild type, spv-1(1498), rhgf-1(ok880), and spv-1(1498);rhgf-1(ok880), measured as the ratio of the width of the distal quarter of the spermatheca (W1) to the width of the proximal quarter of the spermatheca (W2) before the embryo starts to transit out. N ≥ 13. Error bars are mean ± SEM. Statistical comparisons were performed by Brown-Forsythe ANOVA-Dunnett’s T3 multiple comparisons test. (G) Successful embryos transit through the spermatheca of wild-type, spv-1(1498), rhgf-1(ok880), and spv-1(1498);rhgf-1(ok880) worms. N ≥ 14. Statistical comparisons were performed by individuals’ chi-square tests. Stars designate statistical significance (**** P < 0.0001, *** P < 0.001, ** P < 0.01, ns P > 0.05). N, number of samples analyzed.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal