Worm Breeder's Gazette 11(2): 92

These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.

Isolation fo Thermotaxis-defective Mutants

Ikue Mori, Hiroshi Kagoshima and Yasumi Ohshima

Hedgecock and Russell (1975; Proc.  Natl.  Acad.  Sci.  USA 72: 4061)
reported thermotaxis of C.  elegans that can be observed in two ways: 
1) when placed on a linear thermal gradient, the animals migrate 
toward a temperature at which they were cultivated; 2) when placed on 
a radial thermal gradient, the animals move along their adaptive 
growth temperature, thereby making isothermal circular tracks.  By 
screening F2 progeny of EMS-treated N2 for animals which migrated 
abnormally in a linear thermal gradient, Hedgecock and Russell (1975) 
isolated 6 ttx (thermotaxis-defective) mutants.  We are interested in 
identifying and analyzing genes involved in thermotaxis of C.  elegans,
and have begun isolating more ttx mutants with the modification of 
the procedure developed by Hedgecock and Russell (1975).
We noticed earlier that it was difficult to assay reproducibly the 
thermotaxis of N2 strain.  We thought that normal chemotactic 
behaviors of the N2 animals may cause poor or vague thermotaxis, and 
mutagenized CB1377(daf-6(e1377)) animals, which are che-, 
tive due to structural defect in 
amphidial sheath cells (Albert et al., 1981; J.  Comp.  Neurol.  198: 
435), but show normal thermotaxis reproducibly.  F2 progenies of 170 
EMS-treated CB1377 animals were grown at either 15 C or 20  C, and 
were screened for animals which, after placed on a linear thermal 
gradient for 1hr, migrated from their growth temperature toward higher 
(from 15 C to 20 C, or from 20 C to 25 C) or lower (from 20 C to 15 C) 
temperature.  Animals which migrated abnormally were picked among F2 
progenies of 46 mutagenized parents, and thermotaxis of the isolated 
clones was assayed for several generations.  We so far isolated 15 
independent ttx mutants.  Those include thermophilic (2 lines), 
cryophilic (7 lines), and atactic (6 lines) phenotypes, based on 
thermotactic behavior in the linear gradient.  They can be further 
divided into several classes, based on whether or not the mutants are 
capable of making isothermal tracks on a radial thermal gradient.  We 
are in process of mapping these ttx mutations.  At present, we only 
know that ks5, showing cryophilic phenotype on two different 
thermotaxis assays, is probably linked to LGX.
One of our ttx mutations, ks14, exhibits an interesting phenotype: 
it is atactic on the linear gradient, but has a leaky cryophilic 
phenotype on the radial gradient.  In our new assay procedure using 
thin NGM square plate (14 x 10cm) with the linear gradient, individual 
ks14 animals grown at 20 C are capable of making isothermal tracks in 
a range of 17-22 C, whereas parental CB1377 animals grown at 20 C make 
sharp isothermal tracks around 20 C.