Fig 4.
lim-4 is expressed in and acts in the SMB neurons and its expression is autoregulated. (A) GFP expression of oyIs35 animals carrying an integrated
lim-4p::gfp reporter is overlapped with expression of the
flp-12p::mCherry reporter (the SMB marker) but not with expression of the
ceh-17p::dsRed reporter (the SIA marker). Images are derived from z-stacks of confocal microscopy images while images in the upper-left boxed regions are single focal plane confocal microscopy images. Anterior is to the left. Scale bar: 50 um. (B) In
lim-4 mutants, expression of the
lim-4p::gfp reporter is abolished in the cell bodies and processes of the SMB neurons in adult but not L1 larval stage animals. GFP expression of the
lim-4p::gfp reporter is shown in wild-type(left column) or
lim-4(
lsk5) mutant (right column) animals at L1 larval (top) or adult stage (bottom). Note that the
lim-4p::gfp reporter is not expressed in the AWB neurons of
lim-4 mutants [14]. Images are derived from z-stacks of confocal microscopy images. Quantitative analysis of these phenotypes is shown in Table 1. Anterior is to the left. Scale bar: 20 um. (C) Percentage of animals of the indicated genotypes expressing stably integrated
flp-12p:gfp reporter (ynIs25) is shown. Strong, weak or off expression is defined as GFP expression observed at 400x magnification in both cell bodies and processes, in only cell bodies, or not observed either in cell bodies or processes, respectively. Heat shocks were treated to L4 larval stage animals at 33C twice for 30 minutes and after 14 hours, phenotypes were analyzed. Over two independent transgenic lines were tested. n=50 for each. (D) The average of wave width or wavelength of the indicated genotypes. n=30 for each. Error bars are the SEM. *** indicates significantly different between indicated animals at p<0.001 (one-way ANOVA test followed by the Tukey post-hoc test). Heat shocks were applied to L4 larval stage animals at 33C twice for 30 minutes and phenotypes were analyzed after 14 hours. (E-F) Shown are representative pictures of
lim-4 mutants containing the hsp::lim-4cDNA transgene with no heat shock or after heat shock treatment. Images are derived from z-stacks of confocal microscopy images (E: Scale bar: 50 um) and derived from a light microscopy image (F: Scale bar: 0.5 mm). (G)
lim-4 is required to maintain function of the SMB neurons. The average wave width was analyzed in
lim-4(
ky403);Ex[hsp::lim-4cDNA] at 1 day after, 3 day after or 6 day after heat shock treatment. n=30 for each. Error bars are the SEM. *** Significantly different between no heat shock and heat shock treatment conditions at p<0.001(one-way ANOVA test followed by the Tukey post-hoc test).