Dauer larva formation is mainly promoted by a dauer-inducing pheromone and prevented by food supply, but is also influenced by a temperature. Mutations that cause constitutive dauer formation, even in the absence of pheromone, have been isolated and nearly all of these mutations including null mutations are temperature-sensitive: dauer-constitutive (daf-c) mutants grow to adult stage at 15-degree, whereas they arrest at dauer stage at 25-degree. Recently, Hobert et al. (1997) reported that thermotaxis-defective
ttx-3 mutation affects the temperature-sensitive daf-c phenotype of
daf-7 mutants, by enhancing dauer formation at 15-degree and recovery from dauer at 25-degree (1). The
ttx-3 gene encodes a LIM homeodomain protein that is expressed exclusively in AIY, an amphid interneuron essential for thermotaxis (1,2). Thus, sensory signal transduction pathway used for thermotaxis participates at least in part temperature-sensitive daf-c phenotype. Since the
daf-7 gene encodes a TGF-beta-like ligand (3,4), we have begun to investigate whether other daf-c mutations that disrupt TGF-beta-like signaling pathway for dauer/non-dauer developmental decision interact with
ttx-3 or
ttx-1 mutation. The
ttx-1 mutation causes animals to seek cold temperatures as does
ttx-3 (cryophilic phenotype), and is likely to disrupt the function of AFD, a thermosensory neuron required for thermotaxis (2; see Sasakura et al., this issue). We constructed the
daf-1(
m40);
ttx-1(
p767) double mutant and compared it with the
daf-1 single mutant in dauer formation and its recovery. At 25-degree, all
daf-1;
ttx-1 mutant animals initially entered into dauer stage, but 30% of the animals recovered from dauers under uncrowded condition (10-20 animals/plate). By contrast, 100% of
daf-1 animals entered into and stayed at dauer stage at 25-degree under the same uncrowded condition. At 15-degree, 35% of
daf-1;
ttx-1 animals entered into dauer stage, whereas 10% of
daf-1 animals became dauers. The similar results were obtained with
daf-7(
e1372);
ttx-3(
ks5) double mutants that we constructed in this study. About 30% of
daf-7;
ttx-3 animals recovered from dauers at 25-degree, although 100% of
daf-7 animals stayed as dauers. At 15-degree, about 10% of
daf-7;
ttx-3 animals and only 1% of
daf-7 animals became dauers. The
ttx-1 mutant is known to be hypersensitive to dauer-inducing pheromone (5). We thus also investigated dauer formation/recovery of
daf-1;
ttx-1 mutants under crowded condition (150-250 animals/plate). The results showed that dauer formation is more promoted under crowded condition and is significantly enhanced in
daf-1;
ttx-1 animals: 60%, 90% and 100% of double mutants entered into dauer stage at 15-degree, 20-degree and 25-degree, respectively, whereas 25%, 55% and 100% of
daf-1 animals entered into dauer stage at 15-degree, 20-degree and 25-degree, respectively. Taking advantage of the interaction with daf mutations, we are in the process of isolating new thermotaxis-defective mutations that interfere the functions of neurons found to be essential for thermotaxis. We thank Oliver Hobert and Kotaro Kimura for invaluable advice on dauer assays. (1) Hobert et al., 1997, Neuron 19, 345-357. (2) Mori and Ohshima, 1995, Nature 376, 344-348. (3) Ren et al., 1996, Science 274, 1389-1391. (4) Schackwitz et al., 1996, Neuron 17, 719-728. (5) Golden and Riddle, 1984, PNAS 81, 819-823.