Figure 3
Gonadal growth and vulva development phenotypes of pan-1(RNAi) hermaphrodites. (A) Graph showing the gonadal growth scores of pan-1 and control RNAi animals. The number of animals scored for each experiment is indicated. Also shown is RT-PCR analysis of rrf-1(pk1417) cultures (harvested at mid-L4 stage) subjected to control or pan-1RNAi. DIC micrographs of control RNAi (B, C) and pan-1 RNAi (D–G) animals. In images (B-G), the thin arrow indicates vulva, thick arrow indicates distal tip of gonad, and asterisk developing gametes. (B) Control RNAi hermaphrodite at 54 hrs post-feeding. (C) Control RNAi mid-L4 hermaphrodite (38–40 hrs post-feeding). Note luminal stage of vulval development and degree of gonadal growth. (D) pan-1(RNAi) hermaphrodite at 54 hrs post-feeding exhibiting gonadal growth score of 1 and arrested vulva development at the luminal stage. (E) pan-1(RNAi) hermaphrodite at 54 hrs post-feeding with a gonadal growth score of 3 (distal end not shown) and producing gametes. Vulva remains at the luminal stage of development. (F) rrf-1(−); pan-1(RNAi) adult hermaphrodite exhibiting a normal vulval morphology and a morphologically abnormal but differentiated somatic gonad. (G) rrf-1(−); pan-1(RNAi) adult hermaphrodite exhibiting a protruding vulva, a gonadal growth score of 1 and absence of somatic gonad tissue. Both of the animals in (F) and (G) completed the L4 to adult molt and synthesized lateral alae. Epifluorescence (H-J) and corresponding DIC micrographs of AJM-1:GFP expression in control (H) and pan-1 RNAi hermaphrodites (I, J). The animal in (I) exhibits an absence of spermathecal tissue (thick arrow) while the animal in (J) exhibits an abnormally formed and small spermatheca. Epifluorescence micrographs (K, L) of nuclear localized NHR-6:GFP expression in control (K) and pan-1 RNAi hermaphrodites (L). Normal spermathecae in late larvae and adults have 24 cells (thick arrow). In the pan-1(RNAi) animal one gonad arm has two expressing nuclei while the other has a single expressing nucleus. Scale bars = 10 μm.