Figure 2.
Enhancer regions upstream of pezo-1a–h isoforms start codon drove GFP expression in gonad, digestive tract, and associated tissues. (A) A 5-kb promoter upstream of the pezo-1 start codon was used to drive expression of GFP targeting long isoform (a–h) expression. (B) GFP was expressed in pharyngeal gland cells (dorsal and ventral g1) and the AMso glia. (C) GFP was also present in ventral pharyngeal gland cells (vg2), the intestinal valve, and the AMsh glia (D). (E) Cells with anatomy and location consistent with the pharyngeal neuron I4 were labeled. (F) The pharyngeal neuron M1, body wall musculature, and intestine also expressed GFP. Pharyngeal muscles including the isthmus and terminal bulb (pm3 and pm5–8) were labeled in most preps. (G) In the midbody, we found GFP expression in the uterus and vulva, as well as in the HSN neuron. The AG467 translational reporter strain tagging the C-terminus of pezo-1 proteins (PEZO-1::mScarlet) shows PEZO-1 protein accumulation in the vulva and vulval muscles (i). (H) GFP expression was detected in several cells associated with defecation, including cells with anatomy and location consistent with K, F, B, U, Y, and rectD. (I) The translational reporter strain targeting the C domain (shared between all isoforms) showed expression consistent with our transcriptional reporters, with obvious protein accumulation in regions consistent with the glial terminal processes (a), pharyngeal glands (b), and pharyngeal grinder (c). Animals are presented in sagittal views (dorsal up, anterior left) with the exception of F (partial coronal projection). All scale bars are 10 μm unless otherwise indicated. Observations reported based on N = 21 extrachromosomal (transcriptional reporter) and N = 6 integrated (translational reporter) animals. All images obtained using a Leica SP8 white light laser confocal microscope.