The CMK-1 CaMKI and the TAX-4 Cyclic Nucleotide-Gated Channel Regulate Thermosensory Neuron Gene Expression and Function in C. elegans  John S. Satterlee, William S. Ryu, Piali Sengupta  Current Biology  Volume 14, Issue 1, Pages 62-68 (January 2004) DOI: 10.1016/j.cub.2003.12.030

Figure 1 Mutations in cmk-1 and tax-2/4 Affect Gene Expression in the AFD Neurons (A) Expression of stably integrated gcy-8::gfp in wild-type (left) and cmk-1(oy21) (right) adult animals. Arrow points to the cell body of an AFD neuron. Lateral view; scale, 10 μm. (B) Percentage of animals of the indicated genotypes expressing gcy-8::gfp or nhr-38::gfp grown at 25°C (left) or 15°C (right). Strong expression is defined as levels of expression that allow the visualization of the cell bodies and dendrites of the AFD neurons at 100× magnification; weak expression is defined as levels of expression that allow the visualization of the cell bodies but not the dendrites at 100× magnification; no expression is defined as no visible GFP expression at 100× magnification. Adult animals were examined; n > 100 for each. Current Biology 2004 14, 62-68DOI: (10.1016/j.cub.2003.12.030)

Figure 2 Genomic Structure and Expression Pattern of cmk-1 (A) Phylogram showing a neighbor-joining tree of CMK-1 and related protein kinases. The sequences were aligned and analyzed using Bonsai (http://calliope.gs.washington.edu/software/bonsaiWebDocs/BonsaiDocIndex.html). One thousand bootstrap replicates were performed, and bootstrap values are indicated by hatchmarks as follows: (/), <50%; (//), 50%–79%; (///), 80%–100%. HsCDC2 and HsMLCK represent outliers in this analysis. Hs, Homo sapiens; Mm, Mus musculus; Xl, Xenopus laevis; Dm, Drosophila melanogaster; Ce, Caenorhabditis elegans. (B) Exon/intron structure of cmk-1. Filled, hatched, and stippled boxes indicate the extent of the kinase catalytic domain, the ATP binding domain, and the regulatory domains, respectively. The regulatory domain includes the autoinhibitory and the CaM binding domains. The positions and molecular identities of the oy20 and oy21 mutations are indicated. (C) Expression of a cmk-1::gfp fusion gene in an L1 hermaphrodite larva. Strong expression can be seen in the nerve ring (arrow) and along the ventral nerve cord (arrowheads). Anterior is at left; scale, 10 μm. (D) Expression of a gfp-tagged cmk-1 fusion gene under the ttx-1 promoter in adult animals. The ttx-1 promoter drives expression exclusively in the AFD neurons [13]. Arrow points to the cell body of an AFD neuron showing nuclear exclusion of ttx-1::cmk-1::gfp. Anterior is at left; scale, 10 μm. Current Biology 2004 14, 62-68DOI: (10.1016/j.cub.2003.12.030)

Figure 3 TAX-4 Function Is Required during Larval Stages to Regulate gcy-8::gfp Expression Animals of the indicated genotypes carrying stably integrated copies of gcy-8::gfp (oyIs17) were either raised continuously at 15°C or 25°C or shifted to either 25°C or 15°C, respectively, at different developmental stages. The gcy-8::gfp expression pattern was examined in adult animals 3 days after the L4/adult transition. Wild-type levels were defined as levels of expression that enabled the visualization of the cell bodies and dendrites of the AFD neurons at 100× magnification. Data shown are from at least two independent experiments. >100 animals were examined at each time point. Current Biology 2004 14, 62-68DOI: (10.1016/j.cub.2003.12.030)

Figure 4 Thermosensory Behavioral Responses of cmk-1 and tax-4 Mutants Animals of the indicated genotypes were raised at either 17°C (left) or 25°C (right) and subjected to rising temperatures of 19°C → 21°C or falling temperatures of 21°C → 19°C. The turning ratio is defined as the number of turns exhibited by animals in rising temperatures divided by the number of turns exhibited in falling temperatures. Four to six individual animals of each genotype were subjected to up to five alternating upramps and downramps each. Asterisks represent responses that are different from those of wild-type animals at p < 0.05. Current Biology 2004 14, 62-68DOI: (10.1016/j.cub.2003.12.030)