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WBPicture0000012457 Description: Figure S3 Expression of ionotropic ACh receptor genes in the male tail. Muscle abbreviations: DSP: dorsal spicule protractor; VSP: ventral spicule protractor; DSR: dorsal spicule retractor; VSR: ventral spicule retractor; ADP: anal depressor; GER: gubernacular erector; GRT: gubernacular retractor; AOB: anterior oblique; POB: posterior oblique; SPH: sphincter. To obtain the expression pattern of unc-29 in the C. elegans male, we initially fused to the YFP gene a 2.2 kb sequence, which includes a 1.3 kb of sequence upstream of the unc-29 start codon and the genomic sequence of the gene up to the second intron (Punc- 29:YFP). This construct is expressed in the anal depressor muscle and the spicule protractor muscles in the male spicule circuit, and it is also expressed in the body wall muscles, some ventral cord neurons and some head neurons. This is consistent with the previous published expression pattern of unc-38, which encodes an a-subunit that forms the L-AChR with unc-29 (Liu et al. 2007). Nonetheless, the unc-29 genomic sequence also contains important information to facilitate transcription of the gene. Therefore, the expression construct that contained the unc-29 genomic sequence(Punc-29:unc-29::YFP) provided a broader expression pattern than Punc-29:YFP. (A-D) Expression of Punc-29:YFP (yellow) and Pgar- 3:CFP (cyan) imaged from the tail of a same L4 larval male. The two images are merged to show that unc-29 is not expressed in the SPC neurons where gar-3 is expressing (C). The Nomarski image of the tail is also shown (D). Scale bar, 10 um. (E, F) Expression of Punc-29:YFP in the male tail. Scale bar, 20 um. We fused the 2.4 kb sequence upstream of the unc-63 start codon, including the first three codons, to the YFP gene. We found that this construct was expressed in the body wall muscles, the sexually dimorphic anal depressor muscle and sphincter muscle, and every male-specific sex muscle, including the spicule protractor and retractor muscles, the diagonal muscles, the gubernacular muscles, and the oblique muscles. However, it was also expressed in the SPC neurons where unc-29 was not expressed (G-J). When we made an unc-38 promoter-genomic DNA fusion construct, we found that, similarly to unc-63, it was also expressed in every male-specific muscle and the SPC neurons (K-N). (G-J) Expression of Punc-63:YFP in the male tail. Scale bar, 10 um. (K-N) Expression of Punc-38:unc- 38::YFP in the male tail. Scale bar, 10 um. (O, P) Expression of Pacr-12:acr-12::YFP in the male tail. Scale bar, 10 um. (Q-S) Expression of Pacr-18:ChR2::YFP in the male tail. Scale bar, 10 um. (T-V) Expression of Pacr-18:ChR2::YFP (yellow) and Ppkd- 2:CFP (cyan) imaged from the tail of a same adult male. The two images are merged to show that acr-18 is expressed in the HOA neuron that is next to the HOB neuron where pkd-2 is expressing (V). Scale bar, 10 um.
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  Acknowledgment: Template: Reprinted from <Journal_URL>, <Article_URL>. <Publisher_URL> <Publication_year>.
    Publication_year: 2011
    Article_URL: DOI id 10.1371/journal.pgen.1001326
    Journal_URL: PLoSGenetics
    Publisher_URL: PLoS
  Reference: WBPaper00038246