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First published online 21 June 2006
doi: 10.1242/dev.02476

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An antagonistic role for the C. elegans Schnurri homolog SMA-9 in modulating TGFß signaling during mesodermal patterning

¹ Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.
² Department of Embryology, Carnegie Institution of Washington, 3520 San Martin Drive, Baltimore, MD 21218, USA.
³ Departments of Pathology and Genetics, Stanford University School of Medicine, 300 Pasteur Drive-L235, Stanford, CA 94305, USA.

View larger version (80K): [in a new window]   Fig. 1. sma-9 mutations cause a loss of dorsal, and duplication of ventral, M lineage descendants. (A-D) Wild-type (A,C) and sma-9(cc604) (B,D) animals at the L2 (A-B) and adult stage (C-D). Blue arrows: SMs labeled with hlh-8::gfp. Red arrowheads: M-derived CCs labeled with intrinsic CC::gfp. C. elegans also contains four embryonically derived CCs located in the anterior half of the animal (labeled by CC::gfp). Blue arrowheads indicate VM1s (descendants of SMs) labeled with egl-15::gfp. Asterisks indicate the position of the vulva. All images were taken at the same magnification and are lateral views of the animal with anterior towards the left and dorsal upwards. Two SMs (one out of focus) and two M-derived CCs are present in wild type (A,C); four SMs (B) and extra SM descendants (D) are present in sma-9(cc604) but there is a lack of M-derived CCs (B,D). sma-9(cc604) animals (B,D) are smaller than wild-type animals (A,C) at the corresponding stages. (E,F) Schematic representation of the wild-type (E) and sma-9 (F) M lineages. The corresponding stages referred to in the paper are indicated on the right of E. Open circles indicate uterine muscles; filled circles indicate vulval muscles. Lines with no designation indicate bodywall muscles.

View larger version (29K): [in a new window]   Fig. 2. Rescue of sma-9 mutant phenotypes by sma-9 genomic-cDNA hybrid constructs. (A) Schematic diagrams of the predicted full-length sma-9 genomic region and the three genomic-cDNA sma-9 hybrid constructs generated, with exons 1, 4, 6, 9 and 20 labeled. Diagrams are drawn roughly to scale. Yellow ovals represent zinc fingers. The gray box indicates the unique 70 amino acid domain of C2. Broken red outline highlights regions containing genomic sequences. Broken blue outline highlights regions containing cDNA sequences. The broken green line indicates the region targeted by RNAi using pJKL619 (RNAi-N). The broken purple line indicates the region targeted by RNAi using yk127d10 (RNAi-C). Positions of cc606 and cc604 mutations are shown, as well as the region deleted in the tm572 allele. (B) Rescue by sma-9 genomic-cDNA hybrid constructs. All constructs were injected into sma-9(cc604) animals to test for rescue. Rescue was scored using CC::GFP in at least 50 F1 transgenic animals and verified in multiple transgenic lines. +++, at least 50% rescuing efficiency; ++, 25%-50% rescuing efficiency; -, no rescue.

View larger version (86K): [in a new window]   Fig. 3. SMA-9 is localized to nuclei of a wide variety of cell types including the M lineage descendants. (A) A wild-type animal carrying jjEx[3.7kb sma-9p::sma-9C1::gfp::unc-54 3' UTR] immunostained with anti-GFP antibodies. (B-D) A sma-9(cc604) animal at the 2-M stage carrying jjEx[3.7kb sma-9p::sma-9C1::gfp::unc-54 3' UTR] was immunostained with the M lineage-specific anti-MLS-2 antibodies (B) and anti-GFP antibodies (C). (D) Merge of B (red) and C (green). Arrows indicate the two M daughter cells.

View larger version (83K): [in a new window]   Fig. 4. Genetic interactions between sma-9 and TGFß pathway mutants. Phenotypes (A-C) and the corresponding schematic diagrams (A'-C') of the M lineage in sma-3(jj3) (A,A'), sma-3(jj3); sma-9(cc604) (B,B') and sma-3(jj3); sma-9(cc604); jjEx[hlh-8p::sma-3::unc-54 3' UTR+pRF4] (C,C') animals. Red arrowheads indicate M-derived CCs labeled with intrinsic CC::gfp (A-C). Asterisk indicates position of the vulva. All animal views are lateral with anterior towards the left and dorsal upwards.