Figure 5. Polymorphisms in the mechanotransduction TRP-4 channel contribute to the variation of spatial pattern selectivity.
(A) A cartoon illustrating the amino acid substitutions (M36I, G41S, and E67G) in HW TRP-4. (B) Chromosomes I and X were linked to altered spatial pattern selectivity between N2 and HW strains. Hybrid N2/HW strains carrying one HW chromosome (red) and five N2 chromosomes (blue) were quantified for spatial preference at 60 min. n = 12 for N2 and HW worms; n = 10 for worms with chromosome substitutions. (C) Recombinant inbred strains harboring N2 and HW trp-4 genes showed distinct behavior in the spatial pattern preference test at 60 min. n = 12. Estimated regions of recombination (around 13.7 Mb in BJH208 and 14.1 Mb in BJH212) are indicated in the upper panel. The BJH208 strain that carries the N2 trp-4 gene shows enrichment, but the BJH212 strain with HW trp-4 does not (lower panel). (D) Transgenic trp-4(ok1605) worms expressing single-copy N2 or HW TRP-4 in dopaminergic neurons (Pdat-1::trp-4 cDNA) were tested in the spatial pattern preference assay. n = 12 for N2 and trp-4(ok1605) worms, n = 10 for worms with the transgenes. ***p<0.001, when compared to N2; ###p<0.001 and #p<0.05 when compared to trp-4(ok1605) mutants. Error bars denote s.e.m. (E) Transgenic HW worms carrying extrachromosomal arrays with either N2 or HW trp-4 cDNA driven by Pdat-1 were examined in the spatial pattern preference assay. n = 12. Data were collected 60 min after worm loading (unless specified otherwise). One-way ANOVA and Dunnett's multiple comparison tests were used for statistic analyses in Figure 5. ***p<0.001 when compared to N2; ##p<0.01 when compared to HW worms. Error bars denote s.e.m.
Figure 5—figure supplement 1. Both N2 and HW trp-4 cDNAs rescue defects in the swim-to-crawl transition of trp-4(ok1605) mutant worms.
