{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T22:59:13Z","timestamp":1791413953580,"version":"4.3.3"},"reference-count":29,"publisher":"The Company of Biologists","issue":"4","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2005,2,15]]},"abstract":"<jats:p>Mutations in a conserved non-coding region in intron 5 of the Lmbr1 locus, which is 1 Mb away from the sonic hedgehog(Shh) coding sequence, are responsible for mouse and human preaxial polydactyly with mirror-image digit duplications. In the mouse mutants,ectopic Shh expression is observed in the anterior mesenchyme of limb buds. Furthermore, a transgenic reporter gene flanked with this conserved non-coding region shows normal polarized expression in mouse limb buds. This conserved sequence has therefore been proposed to act as a long-range,cis-acting regulator of limb-specific Shh expression. Previous phylogenetic studies have also shown that this sequence is highly conserved among tetrapods, and even in teleost fishes. Paired fins of teleost fishes and tetrapod limbs have evolved from common ancestral appendages, and polarized Shh expression is commonly observed in fins. In this study, we first show that this conserved sequence motif is also physically linked to the Shh coding sequence in a teleost fish, the medaka, by homology search of a newly available genomic sequence database. Next, we show that deletion of this conserved intronic sequence by targeted mutation in the mouse results in a complete loss of Shh expression in the limb bud and degeneration of skeletal elements distal to the stylopod\/zygopod junction. This sequence contains a major limb-specific Shh enhancer that is necessary for distal limb development. These results suggest that the conserved intronic sequence evolved in a common ancestor of fishes and tetrapods to control fin and limb development.<\/jats:p>","DOI":"10.1242\/dev.01613","type":"journal-article","created":{"date-parts":[[2005,1,27]],"date-time":"2005-01-27T17:28:07Z","timestamp":1106846887000},"page":"797-803","source":"Crossref","is-referenced-by-count":361,"title":["Elimination of a long-range cis-regulatory module causes complete loss of limb-specific<i>Shh<\/i>expression and truncation of the mouse limb"],"prefix":"10.1242","volume":"132","author":[{"given":"Tomoko","family":"Sagai","sequence":"first","affiliation":[{"name":"Mammalian Genetics Laboratory, National Institute of Genetics, Yata-1111 Mishima, Shizuoka-ken 411-8540, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masaki","family":"Hosoya","sequence":"additional","affiliation":[{"name":"Mammalian Genetics Laboratory, National Institute of Genetics, Yata-1111 Mishima, Shizuoka-ken 411-8540, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Youichi","family":"Mizushina","sequence":"additional","affiliation":[{"name":"Mammalian Genetics Laboratory, National Institute of Genetics, Yata-1111 Mishima, Shizuoka-ken 411-8540, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masaru","family":"Tamura","sequence":"additional","affiliation":[{"name":"Mammalian Genetics Laboratory, National Institute of Genetics, Yata-1111 Mishima, Shizuoka-ken 411-8540, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toshihiko","family":"Shiroishi","sequence":"additional","affiliation":[{"name":"Mammalian Genetics Laboratory, National Institute of Genetics, Yata-1111 Mishima, Shizuoka-ken 411-8540, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","reference":[{"key":"2021042605232386400_REF1","doi-asserted-by":"crossref","unstructured":"Boffelli, D., Nobrega, M. A. and Rubin, E. M.(2004). Comparative genomics at the vertebrate extremes. Nat. Rev. Genet.5,456-465.","DOI":"10.1038\/nrg1350"},{"key":"2021042605232386400_REF2","doi-asserted-by":"crossref","unstructured":"Buscher, D., Bosse, B., Heymer, J. and Ruther, U.(1997). Evidence for genetic control of Sonic hedgehog by Gli3 in mouse limb development. Mech. Dev.62,175-182.","DOI":"10.1016\/S0925-4773(97)00656-4"},{"key":"2021042605232386400_REF3","doi-asserted-by":"crossref","unstructured":"Charite, J., McFadden, D. G. and Olson, E. N.(2000). The bHLH transcription factor dHAND controls Sonic hedgehog expression and establishment of the zone of polarizing activity during limb development. Development127,2461-2470.","DOI":"10.1242\/dev.127.11.2461"},{"key":"2021042605232386400_REF4","doi-asserted-by":"crossref","unstructured":"Chiang, C., Litingtung, Y., Harris, M. P., Simandl, B. K., Li,Y., Beachy, P. A. and Fallon, J. F. (2001). Manifestation of the limb prepattern: limb development in the absence of sonic hedgehog function. Dev. Biol.236,421-435.","DOI":"10.1006\/dbio.2001.0346"},{"key":"2021042605232386400_REF5","doi-asserted-by":"crossref","unstructured":"Chiang, C., Litingtung, Y., Lee, E., Young, K. E., Corden, J. L., Westphal, H. and Beachy, P. A. (1996). Cyclopia and defective axial patterning in mice lacking Sonic hedgehog gene function. Nature383,407-413.","DOI":"10.1038\/383407a0"},{"key":"2021042605232386400_REF6","doi-asserted-by":"crossref","unstructured":"Grandel, H. and Schulte-Merker, S. (1998). The development of the paired fins in the zebrafish (Danio rerio). Mech. Dev.79,99-120.","DOI":"10.1016\/S0925-4773(98)00176-2"},{"key":"2021042605232386400_REF7","doi-asserted-by":"crossref","unstructured":"Harrow, F., Amuta, J. U., Hutchinson, S. R., Akwaa, F. and Ortiz, B. D. (2004). Factors binding a non-classical Cis-element prevent heterochromatin effects on locus control region activity. J. Biol. Chem.279,17842-17849.","DOI":"10.1074\/jbc.M401258200"},{"key":"2021042605232386400_REF8","doi-asserted-by":"crossref","unstructured":"Heutink, P., Zguricas, J., Oosterhout, L. v., Breedveld, G. J.,Testers, L., Sandkuijl, L. A., Snijders, P. J. L. M., Weissenbach, J.,Lindhout, D., Hovius, S. E. R. et al. (1994). The gene for triphalangeal thumb maps to the subtelomellic region of chromosome 7q. Nat. Genet.6,287-292.","DOI":"10.1038\/ng0394-287"},{"key":"2021042605232386400_REF9","doi-asserted-by":"crossref","unstructured":"Ianakiev, P., van Baren, M. J., Daly, M. J., Toledo, S. P.,Cavalcanti, M. G., Neto, J. C., Silveira, E. L., Freire-Maia, A., Heutink, P.,Kilpatrick, M. W. et al. (2001). Acheiropodia is caused by a genomic deletion in C7orf2, the human orthologue of the Lmbr1 gene. Am. J. Hum. Genet.68,38-45.","DOI":"10.1086\/316955"},{"key":"2021042605232386400_REF10","unstructured":"Jegalian, B. G. and De Robertis, E. M. (1992). Homeotic transformations in the mouse induced by overexpression of a human Hox3.3 transgene. Cell71,901-910."},{"key":"2021042605232386400_REF11","doi-asserted-by":"crossref","unstructured":"Johnson, R. L. and Tabin, C. J. (1997). Molecular models for vertebrate limb development. Cell90,979-990.","DOI":"10.1016\/S0092-8674(00)80364-5"},{"key":"2021042605232386400_REF12","doi-asserted-by":"crossref","unstructured":"Kraus, P., Fraidenraich, D. and Loomis, C. A.(2001). Some distal limb structures develop in mice lacking Sonic hedgehog signaling. Mech. Dev.100, 45-58.","DOI":"10.1016\/S0925-4773(00)00492-5"},{"key":"2021042605232386400_REF13","doi-asserted-by":"crossref","unstructured":"Lettice, L. A., Heaney, S. J., Purdie, L. A., Li, L., de Beer,P., Oostra, B. A., Goode, D., Elgar, G., Hill, R. E. and de Graaff, E.(2003). A long-range Shh enhancer regulates expression in the developing limb and fin and is associated with preaxial polydactyly. Hum. Mol. 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Transient establishment of anteroposterior polarity in the zebrafish pectoral fin bud in the absence of sonic hedgehog activity. Development126,4817-4826.","DOI":"10.1242\/dev.126.21.4817"},{"key":"2021042605232386400_REF20","doi-asserted-by":"crossref","unstructured":"Riddle, R. D., Johnson, R. L., Laufer, E. and Tabin, C.(1993). Sonic Hedgehog mediates the polarizing activity of ZPA. Cell75,1401-1416.","DOI":"10.1016\/0092-8674(93)90626-2"},{"key":"2021042605232386400_REF21","doi-asserted-by":"crossref","unstructured":"Ruvinsky, I. and Gibson-Brown, J. J. (2000). Genetic and developmental bases of serial homology in vertebrate limb evolution. Development127,5233-5244.","DOI":"10.1242\/dev.127.24.5233"},{"key":"2021042605232386400_REF22","doi-asserted-by":"crossref","unstructured":"Sagai, T., Masuya, H., Tamura, M., Shimizu, K., Yada, Y.,Wakana, S., Gondo, Y., Noda, T. and Shiroishi, T. (2004). Phylogenetic conservation of a limb-specific, cis-acting regulator of Sonic hedgehog (Shh). Mamm. Genome15, 23-34.","DOI":"10.1007\/s00335-033-2317-5"},{"key":"2021042605232386400_REF23","doi-asserted-by":"crossref","unstructured":"Sharpe, J., Lettice, L., Hecksher-Sorensen, J., Fox, M., Hill,R. and Krumlauf, R. (1999). Identification of sonic hedgehog as a candidate gene responsible for the polydactylous mouse mutant Sasquatch.Curr. Biol.9, 97-100.","DOI":"10.1016\/S0960-9822(99)80022-0"},{"key":"2021042605232386400_REF24","doi-asserted-by":"crossref","unstructured":"Sordino, P., van der Hoeven, F. and Duboule, D.(1995). Hox gene expression in teleost fins and the origin of vertebrate digits. Nature375,678-681.","DOI":"10.1038\/375678a0"},{"key":"2021042605232386400_REF25","doi-asserted-by":"crossref","unstructured":"Srivastava, D., Cserjesi, P. and Olson, E. N.(1995). A subclass of bHLH proteins required for cardiac morphogenesis. 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