Article Figures & Tables
Figures
- Fig. 1.
Rescue of daf-9 gonadal migration phenotype. (A) Hypodermal expression of daf-9, directed by a dpy-7p::daf-9::GFP transgene, was sufficient to rescue the gonadal migration phenotype of daf-9(e1406) dpy-7(sc27) mutant animals. Shown is a late L4 larva as indicated by the vulval morphology (marked with an asterisk). (B) Expression of daf-9 in a subset of sensory neurons, directed by a che-2p::daf-9::GFP transgene, failed to rescue the gonadal migration phenotype of daf-9(e1406) dpy-7(sc27) mutant animals. Shown is a late L4 larva, as indicated by the vulval morphology (marked with an asterisk).
- Fig. 2.
Partial suppression of daf-2 dauer arrest by daf-9 expression in the hypodermis. (A) A daf-2(e1370) partial dauer animal carrying a dpy-7p::daf-9::GFP transgene. Although the vulva displayed L4 morphology (inset), gonadal migration was arrested and abundant refractile bodies, indicative of lipid accumulation, were apparent in the intestinal cells. The arrow indicates the position of the distal tip cell (DTC). (B) A daf-2(e1370) dauer animal showing arrested vulval development, gonadal migration and refractile bodies in the intestinal cells. All animals were raised at 25°C.
- Fig. 3.
Effect of daf-9 overexpression on life span of daf-2 animals. Adult lifespan of N2 (n=55), daf-2(e1370) (n=54) and daf-2(e1370) Ex[col-12p::daf-9::GFP] (#1, n=104; #2, n=108) were determined at 25°C. Two independent trials were conducted and the result of one trial is shown.
- Fig. 4.
Transcriptional control of daf-9 by daf-12. (A,B) An N2 wild-type larva carrying a daf-9p::daf-9 genomic::GFP transgene at late L2 stage. (A) Normaski image. (B) Fluorescence image showing strong daf-9::GFP expression in the hypodermis. (C,D) A dpy-7(sc27) daf-12(m20) larva carrying a daf-9p::daf-9 genomic::GFP transgene at late L2 stage. (C) Normaski image. (D) Fluorescence image showing daf-9::GFP expression in the XXXL/R cell but not in the hypodermis. (E) Summary of daf-9::GFP expression pattern driven by five transgenes that encompass different promoter, exonic and intronic sequence from the endogenous daf-9 locus. In construct v, a nuclear receptor (NR) consensus half site (underlined) in intron 1 was mutated to a LexA-binding site with the altered bases in grey. Transgenes i, ii and iv rescued the dauer arrest phenotype of daf-9(e1406) dpy-7(sc27) mutant animals. n.d., not determined.
Tables
- Table 1.
Rescue of daf-9(e1406) with extrachromosomal arrays (20°C 68 hours post egg lay)
% transgenic non-dauers (n)* Transgene Line 1 Line 2 -† 0 (>200) - daf-9p::daf-9 genomic::GFP 100 (264) 100 (416) dpy-7p::daf-9 cDNA::GFP 100 (412) 100 (185) sdf-9p::daf-9 cDNA::GFP 97 (320) 98 (557) che-2p::daf-9 cDNA::GFP‡,§ 62 (145) 44 (323) -
↵* The genetic background was daf-9(e1406) dpy-7(sc27).
-
↵† In addition to no transgene control, two control Ex[mec-7::GFP] transgenes did not rescue daf-9(e1406). No Dpy non-dauer progeny from daf-9(e1406) dpy-7(sc27)/szT1 Ex[mec-7p::GFP] was observed (total number of brood examined=37).
-
↵‡ All other transgenic animals not scored as non-dauers were partial dauers.
-
↵§ % transgenic non-dauers at 92 hours post egg lay are: line 1, 79%; line 2, 84%.
-
- Table 2.
Gonadal migration phenotype (20°C 92 hours post egg lay)
% normal gonadal migration (n)*,† Transgene Line 1 Line 2 - 0 (>200) - daf-9p::daf-9 genomic::GFP 100 (>200) 100 (>200) dpy-7p::daf-9 cDNA::GFP 100 (412) 100 (185) sdf-9p::daf-9 cDNA::GFP 97 (311) 100 (544) che-2p::daf-9 cDNA::GFP 34 (110) 12 (292) - Table 3.
Effect of daf-9 overexpression on dauer formation (25°C 55 hours post egg lay)
% transgenic non-dauers (n) Line 1 Line 2 dpy-7p::daf-9 cDNA::GFP daf-7(e1372)† 100 (140) 74 (210)* daf-1(m40)† 100 (124) 100 (122) sdf-9p::daf-9 cDNA::GFP daf-7(e1372)† 0 (101) 0.4 (488) - Table 4.
Hypodermal expression of daf-9::GFP (L2 larvae)
% animals expressing hypodermal daf-9::GFP (n) daf-9p::daf-9 genomic::GFP 15°C 25°C Wild type 93 (136) 96 (80) dpy-7(sc27) daf-12(m20) 19 (247) 5 (127)
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