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| WBPicture0000015228 | Description | Figure 1. Comprehensive analysis of EGL-19 isoform expression:(A) The egl-19 locus is predicted to encode three isoforms of EGL-19 (a, b, and c). Exons are indicated as gray boxes linked by introns. The exon highlighted blue is unique to isoforms b and c. Black boxes represent the locations of the regions encoding the transmembrane domains that form the ion channel pore. Note that the truncated c isoform lacks these transmembrane domains. Fluorescent tags were inserted using CRISPR/Cas9 knock-in at the endogenous egl-19 locus. Colored boxes indicate the insertion location for each fluorescent tag relative to egl-19 gene structure (mSc = mScarlet and mNG = mNeonGreen). Orange box indicates the putative location of a neuron-specific promoter identified by the RegAtlas project (Serizay et al. 2020). Red font refers to RNAseq splice junction data that was obtained from WormBase (version WS288) and analyzed in this study. (B-G) Confocal imaging of EGL-19 tag expression. Yellow arrows indicate overlapping expression of mScarlet and GFP, revealing localization of the EGL-19(b) isoform. Red arrows indicate expression of GFP, revealing localization of the EGL-19(c) isoform. White arrows indicate punctate expression of mScarlet, which we hypothesize represents accumulation of mScarlet::EGL-19 in lysosomes. Pink arrows refer to a neuron which expresses both mNG and GFP, suggesting robust expression of EGL-19(c). Blue arrows refer to a bilateral pair of neurons which express mNG, but not GFP or mScarlet, suggesting more limited expression of EGL-19(c). (B) Representative images of co-expression of mScarlet::EGL-19(ab)/EGL-19(bc)::mNG in the pharyngeal bulb. Animals are F1 heterozygotes obtained from mating homozygous mScarlet::egl-19(ab) and egl-19(bc)::mNG parents. mNG expression in the pharyngeal muscle is so bright that it precludes analysis of mScarlet co-localization (compare mNG in B with GFP in E). (C-G) Representative images of co-expression of mScarlet::EGL-19(ab)/EGL-19(abc)::GFP in several different tissues. Animals are F1 heterozygotes obtained from mating homozygous mScarlet::egl-19(ab) and egl-19(abc)::GFP parents. mScarlet and GFP fluorescence levels are relatively balanced, allowing observation of overlapping expression in the "MERGE" column. | |||
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| Name | ec04311ababaaf71181859d83a2d3725.jpg | ||||
| Depict | Expr_pattern | Expr16371 | |||
| Expr16372 | |||||
| Expr16373 | |||||
| Anatomy | WBbt:0003679 | ||||
| WBbt:0005451 | |||||
| WBbt:0005813 | |||||
| WBbt:0005821 | |||||
| Cellular_component | GO:0005829 | ||||
| GO:0005886 | |||||
| Acknowledgment | Template | Reprinted from <Journal_URL> | |||
| Publication_year | 2023 | ||||
| Article_URL | DOI | id | 10.17912/micropub.biology.000858 | ||
| Journal_URL | microPublicationBiology | ||||
| Publisher_URL | Micropublication | ||||
| Reference | WBPaper00065588 |
