Fig 5. Phosphodiesterase PDE-6 is necessary for normal transit times and Ca²⁺ dynamics.

(A) population assay of wild type animals grown on control RNAi, plc-1(RNAi), pde-1(RNAi), pde-2(RNAi), pde-3(RNAi), pde-4(RNAi), pde-5(RNAi), pde-6(RNAi), pde-12(RNAi), C01B10.9 and T02G6.3. Spermathecae were scored for the presence or absence of an embryo (occupied or unoccupied), presence of a fragment of an embryo (small piece occupied), or the presence of endomitotic oocytes in the gonad arm (emo) phenotypes. The total number of unoccupied spermatheca was compared to the sum of all other phenotypes using the Fisher’s exact t-test. N is the total number of spermathecae counted. Control RNAi was compared to all other RNAi treatments. Entry time (B) and exit time (C) were compared using Fisher’s exact t-test (two dimensional x² analysis). (B,C) Values are color coded corresponding to transit phenotype (purple: successful exit, orange: traps. Representative normalized Ca²⁺ traces and kymograms of control RNAi (D) and pde-6(RNAi) (E) are shown with time of entry, distal neck closure, and time the sp-ut valve opens and closes indicated. Levels of Ca²⁺ signal were normalized to 30 frames before oocyte entry. Kymograms were generated by averaging over the columns of each movie frame (see methods). Refer to S3A Fig and S3G Fig for additional Ca²⁺ traces, and S4A Fig and S9A Fig for additional kymograms. (F-I) Values are color coded corresponding to transit phenotype (purple: successful exit, orange: traps). (F) The number of Ca²⁺ peaks, (G) the peaks per second, and the amount of time after oocyte entry required to reach either (H) half the maximum or (I) maximum Ca²⁺ signal were quantified for wild type animals treated with control and pde-6(RNAi). Values were compared using Fisher’s exact t-test. Stars designate statistical significance (**** p<0.0001, *** p<0.005, ** p<0.01, * p<0.05).