Figure 2.
egl-13 functions cell autonomously to drive O2- and CO2-sensing neuron cell fate. (A) Dorsal view of a young adult hermaphrodite expressing an egl-13prom1::mCherry transcriptional reporter transgene (top panels), a
gcy-33prom::
gcy-33::gfp translational reporter marking BAGL/R, URXL/R, AQR and PQR (center panels) and a merge of the two pictures showing co-localization in the O2- and CO2-sensing neurons (bottom panels). Left hand panels show the head region and right hand panels the tail region. We also observed expression of egl-13prom1::mCherry in muscle and vulval cells (not shown). A: anterior, L: left. The scale bar in lower panel is 20 mm. The egl-13prom1::mCherry transgene is rpEx272 and the
gcy-33prom::GCY-33::gfp transgene is rpIs7. (B) Fluorescence micrographs of
gcy-33prom::
gcy-33::gfp expression in a wild-type animal (top panels), an egl- 13
(ku194) mutant animal (center panels) and an
egl-13(
ku194) mutant animal rescued by transgenic expression of egl-13isoformAcDNA driven by the endogenous 3.5 kb egl-13prom1 (bottom panels). Left hand panels show the head region and right hand panels the tail region. (C) Transgenic expression of egl-13isoformAcDNA under the control of egl-13prom1 rescues
egl-13 mutant neuronal phenotypes, in the
gcy-33prom::
gcy-33::gfp strain. n = 52-55. ***P,0.001. Lines 1-3 = independent transgenic rescue lines. See materials and methods for neuronal scoring criteria used. (D) Transgenic expression of egl-13isoformAcDNA under the control of the
gcy-331kb (BAG-specific) or
unc-86(700bp) (URX-specific) promoters rescues
egl-13 mutant neuronal phenotypes in the
flp-19prom::gfp (BAG) and
gcy-33prom::
gcy-33::gfp (URX) strains. n = 39-55. ***P,0.001. Lines 1-2 = independent transgenic rescue lines. See materials and methods for neuronal scoring criteria used. (E) Transgenic expression of
sox-2 cDNA under the control of egl-13prom1 is unable to rescue
egl-13 mutant neuronal phenotypes, in the
gcy-33prom::
gcy-33::gfp strain. n = 56-60. (n.s.) indicates no significant difference from non transgenic
egl-13 mutant animals. Lines 1-2 = independent transgenic rescue lines. See materials and methods for neuronal scoring criteria used.