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First published online 17 March 2004
doi: 10.1242/dev.01069

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Intercellular signaling of reproductive development by the C. elegans DAF-9 cytochrome P450

Department of Molecular Biology, Massachusetts General Hospital and Department of Genetics, Harvard Medical School, Boston, MA 02114, USA

View larger version (54K): [in a new window]   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).

View larger version (155K): [in a new window]   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.

View larger version (16K): [in a new window]   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.

View larger version (42K): [in a new window]   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.