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First published online 22 November 2005
doi: 10.1242/jcs.02678

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The Caenorhabditis elegans GATA factor elt-1 is essential for differentiation and maintenance of hypodermal seam cells and for normal locomotion

Institute of Comparative Medicine, Faculty of Veterinary Medicine, University of Glasgow, Bearsden Road, Glasgow, G61 1QH, UK

View larger version (75K): [in a new window]   Fig. 1. Expression of elt-1 reporter gene in JG31 vpIs7 transgenic worms. (A) Embryo at approximately 200 minutes of development (pre-lima stage). (B) Comma stage embryo showing GFP expression in the lateral seam cells. (C) L1 showing GFP expression in retrovesicular ganglion. (D) L4 showing lacZ expression in lateral seam cells. (E) L3 and L4 larvae showing lacZ expression in ventral cord neurons. (F) Expression of lacZ in ventral cord neurons of an adult worm. (G) Adult worm with lacZ expression in retrovesicular ganglion and axon encircling the pharynx. (H) Expression of lacZ in vulval muscles. (I) GFP expression in SET cells of an adult male worm. A C. elegans embryo is approximately 50 µm in length. An adult hermaphrodite is approximately 1 mm in length.

View larger version (87K): [in a new window]   Fig. 2. Effects of elt-1 RNAi applied during embryogenesis. (A) Homozygous arrested elt-1(zu180) embryo expressing the ijIS12 dpy-7::GFP transgene. (B) RNAi arrested elt-1 embryo expressing the dpy-7::GFP transgene. (C) RNAi of elt-1 in L1 larvae with Lpy-Dpy phenotype. (D) JR1000 L1 larva showing the wild-type expression pattern of the SCM::GFP and ajm-1::GFP reporter genes. (E) RNAi of elt-1 in Lpy-Dpy JR1000 larva showing loss of SCM::GFP and ajm-1::GFP expression and disorganisation of the remaining seam cells. (F) Normal dpy-7::GFP L1 expression pattern from the ijIS12 transgene. (G) RNAi of elt-1 Lpy-Dpy L1 larvae expressing dpy-7::GFP from the ijIS12 transgene showing loss of expression in positions that should be occupied by seam cells (arrows). (H) Wild-type JG5 L1 larvae showing normal elt-3::GFP expression in the dorsal/ventral hypodermis (but not the lateral seam cells) and cells immediately anterior and posterior to the gut (virL, virR, rect D, rect VL and rect VR) (Gilleard et al., 1999). (I) RNAi of elt-1 Lpy-Dpy L1 larva showing loss of elt-3::GFP expression from the vpIS1 transgene in all the hypodermal cells of the head and dorsal/ventral hypodermis. A C. elegans embryo is approximately 50 µm in length. A newly hatched L1 larva is approximately 240 µm in length.

View larger version (64K): [in a new window]   Fig. 3. Postembryonic elt-1 RNAi phenotype. (A) Adult elt-1 RNAi worm showing herneation of the gonad through the vulva and associated flaccid phenotype. (B) SEM of an adult hermaphrodite vulva. (C) SEM of an adult hermaphrodite vulva following elt-1 RNAi. (D) Timing of onset of the adult phenotype; L1 larvae containing the ksIS12 col-19::GFP transgene were placed on elt-1 RNAi feeding plates. The line graph shows the number of worms expressing col-19::GFP (n=100) and the bar chart shows the number of worms showing the burst-vulva phenotype. An adult C. elegans hermaphrodite is approximately 1 mm in length.

View larger version (33K): [in a new window]   Fig. 4. Seam-cell loss induced by postembryonic elt-1 RNAi. (A) Mean number of SCM::GFP expressing cells in different stages following elt-1 RNAi (unilateral counts, n=10; error bars indicate s.d.). N2 indicates the JR667 strain, wIs51 SCM::GFP in N2 background, and NL indicates the JG147 strain, wIs51 SCM::GFP marker in rrf-3(pk1426) RNAi-hypersensitive mutant background. (B,C) Negative-control JR667 and JG147 L4 larvae, respectively. (D,E) RNAi of elt-1 in JR667 and JG147 L4 larvae respectively. (F) RNAi of elt-1 in JR1000 L4 larva showing loss of expression of both the SCM::GFP and ajm-1::GFP markers. (G) RNAi of elt-1 in JR667 L4 larva with dispersed SCM::GFP expression in a degenerate seam cell (arrow). (H) Mean number (±s.d.) of SCM::GFP expressing cells (unilateral), number of SET cells and number of bursal rays of the male tail after elt-1 RNAi. (I) SEM of a normal male tail. (J) SEM of male tail following elt-1 RNAi. An adult C. elegans hermaphrodite is approximately 1 mm in length.

View larger version (103K): [in a new window]   Fig. 5. Effect of postembryonic elt-1 RNAi on cuticle structure. (A) SEM of a normal adult worm showing lateral alae. (B) SEM of adult worm following postembryonic elt-1 RNAi showing loss of alae. (C) Adult worm expressing col-19::GFP from the kaIs12 transgene showing normal cuticle structure. (D) Adult hermaphrodite showing loss of alae and discontinuity of COL-19::GFP expression following elt-1 RNAi. (E) SEM of an adult hermaphrodite following elt-1 RNAi showing abnormal aggregation of material on the cuticle surface. (F) COL-19::GFP is present in material on the surface of an elt-1 RNAi adult hermaphrodite. An adult C. elegans hermaphrodite is approximately 1 mm in length. Individual cuticular annulae of an adult hermaphrodite are approximately 1 µm in width.

View larger version (61K): [in a new window]   Fig. 6. Effect of postembryonic elt-1 RNAi on dauer larvae formation. (A) Normal daf-2(m577) dauer larvae. (B) RNAi of elt-1 in daf-2(m577) dauer larvae. (C) RNAi of elt-1 in daf-2(m577) dauer larva at 40 hours of development. (D) SEM of normal daf-2(m577) dauer larva. (E) SEM of elt-1 RNAi daf-2(m577) dauer larva taken at 40 hours of development. (F) SEM of an elt-1 RNAi daf-2(m577) dauer at 60 hours of development. (G) SCM::GFP expression in daf-2(m577) dauer larva. (H) SCM::GFP expression in elt-1 RNAi daf-2(m577) dauer larva. (I) Mean number (±s.d.) of SCM::GFP-expressing cells (unilateral, n=10 larvae) in elt-1 RNAi L2d and dauer larvae and negative controls. A C. elegans dauer larvae is approximately 400 µm in length.

View larger version (59K): [in a new window]   Fig. 7. Effect of postembryonic RNAi on locomotion. (A) N2 L4 larvae on negative-control plates. (B) N2 L4 larvae on elt-1 RNAi plates. The increased amplitude of the sinusoidal waveform can be seen by the exaggeratedly `looped' tracks on the bacterial lawn (arrows). (C) Mean values (±s.d.) of the number of head deflections per minute for L4 larvae on elt-1 RNAi and negative-control plates (n=10). Data is shown for the wild type (N2) and RNAi-hypersensitive rrf-3(pk1426) mutants (NL). The differences between the RNAi and control groups were statistically significant by a nonpaired t-test; P=0.0005 for N2 and P=0.0001 for rrf-3(pk1426) mutants. (D) Percentage of worms moving before and after heat-shock-induced ectopic expression of elt-1 or elt-3. JM53 and JM55 carry independent chromosomally integrated transgenes for ectopic expression of elt-1. JM59 and JM60 carry independent chromosomally integrated transgenes for ectopic expression of elt-3. The results show the mean values (±s.d.) for four independent experiments in which 100 worms were counted per group. An adult C. elegans hermaphrodite is approximately 1 mm in length.

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