{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T20:19:15Z","timestamp":1791317955223,"version":"4.3.1"},"reference-count":38,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5369","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[1998,6,5]]},"abstract":"<jats:p>Veratrum alkaloids and distal inhibitors of cholesterol biosynthesis have been studied for more than 30 years as potent teratogens capable of inducing cyclopia and other birth defects. Here, it is shown that these compounds specifically block the Sonic hedgehog (Shh) signaling pathway. These teratogens did not prevent the sterol modification of Shh during autoprocessing but rather inhibited the response of target tissues to Shh, possibly acting through the sterol sensing domain within the Patched protein regulator of Shh response.<\/jats:p>","DOI":"10.1126\/science.280.5369.1603","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:43:20Z","timestamp":1027763000000},"page":"1603-1607","source":"Crossref","is-referenced-by-count":754,"title":["Teratogen-Mediated Inhibition of Target Tissue Response to\n            <i>Shh<\/i>\n            Signaling"],"prefix":"10.1126","volume":"280","author":[{"given":"Michael K.","family":"Cooper","sequence":"first","affiliation":[{"name":"M. K. Cooper, Department of Neurology and Howard Hughes Medical Institute, Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205."},{"name":"J. A. Porter, K. E. Young, P. A. Beachy, Howard Hughes Medical Institute, Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeffery A.","family":"Porter","sequence":"additional","affiliation":[{"name":"M. K. Cooper, Department of Neurology and Howard Hughes Medical Institute, Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205."},{"name":"J. A. Porter, K. E. Young, P. A. Beachy, Howard Hughes Medical Institute, Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keith E.","family":"Young","sequence":"additional","affiliation":[{"name":"M. K. Cooper, Department of Neurology and Howard Hughes Medical Institute, Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205."},{"name":"J. A. Porter, K. E. Young, P. A. Beachy, Howard Hughes Medical Institute, Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philip A.","family":"Beachy","sequence":"additional","affiliation":[{"name":"M. K. Cooper, Department of Neurology and Howard Hughes Medical Institute, Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205."},{"name":"J. A. Porter, K. E. Young, P. A. Beachy, Howard Hughes Medical Institute, Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/383407a0"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0959-4388(98)80004-4"},{"key":"e_1_3_2_4_2","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1038\/ng1196-353","volume":"14","author":"Belloni E.","year":"1996","unstructured":"Belloni E., et al., Nature Genet.14, 353 (1996); E. Roessler et al.,ibid., p. 357.","journal-title":"Nature Genet."},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.7985023"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/374363a0"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.274.5285.255"},{"key":"e_1_3_2_8_2","first-page":"451","volume":"21","author":"Roux C.","year":"1964","unstructured":"Roux C., Arch. Fr. Pediatr.21, 451 (1964); C. R. Soc. Biol. 160, 1353 (1966).","journal-title":"Arch. Fr. Pediatr."},{"key":"e_1_3_2_9_2","doi-asserted-by":"crossref","first-page":"911","DOI":"10.1016\/0006-2952(86)90076-6","volume":"35","author":"Aufenanger J.","year":"1986","unstructured":"D. B. Dehart, L. Lanoue, G. S. Tint, K. K. Sulik, Am. J. Med. Genet. 68, 328 (1997); Aufenanger J., Pill J., Schmidt F. H., Stegmeier K., Biochem. Pharmacol. 35, 911 (1986).","journal-title":"Biochem. Pharmacol."},{"key":"e_1_3_2_10_2","doi-asserted-by":"crossref","first-page":"6230","DOI":"10.1016\/S0021-9258(18)83338-5","volume":"264","author":"Popj\u00e1ck G.","year":"1989","unstructured":"M. Kraml, J. F. Bagli, D. Dvornick, Biochem. Biophys. Res. Commun. 15, 455 (1964); Popj\u00e1ck G., Meenan A., Parish E. J., Nes W. D., J. Biol. Chem. 264, 6230 (1989).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_2_11_2","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1056\/NEJM199401133300205","volume":"330","author":"Tint G. S.","year":"1994","unstructured":"R. I. Kelley, et al., Am. J. Med. Genet. 66, 478 (1996); Tint G. S., et al., N. Eng. J. Med. 330, 107 (1994).","journal-title":"N. Eng. J. Med."},{"key":"e_1_3_2_12_2","doi-asserted-by":"crossref","first-page":"19417","DOI":"10.1074\/jbc.270.33.19417","volume":"270","author":"Stefansson S.","year":"1995","unstructured":"T. E. Willnow, et al., Proc. Natl. Acad. Sci. U.S.A. 93, 8460 (1996); Stefansson S., Chappell D. A., Argraves K. M., Strickland D. K., Argraves W. S., J. Biol. Chem. 270, 19417 (1995).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1002\/tera.1420010103"},{"key":"e_1_3_2_14_2","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1101\/SQB.1997.062.01.025","volume":"62","author":"Beachy P. A.","year":"1997","unstructured":"Beachy P. A., et al., Cold Spring Harbor Symp. Quant. Biol.62, 191 (1997).","journal-title":"Cold Spring Harbor Symp. Quant. Biol."},{"key":"e_1_3_2_15_2","unstructured":"Jervine [250 \u03bcl of a sonicated solution (1 mg\/ml)] was applied to windowed fertile chick eggs (White Leghorn Chestertown MD) at 14 hours of incubation. After 4 days of further incubation the embryos were processed for scanning electron microscopy."},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(93)90248-O"},{"key":"e_1_3_2_17_2","doi-asserted-by":"crossref","first-page":"2537","DOI":"10.1242\/dev.121.8.2537","volume":"121","author":"Marti E.","year":"1995","unstructured":"Marti E., Takada R., Bumcrot D. A., Sasaki H., McMahon A. P., Development121, 2537 (1995).","journal-title":"Development"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1350865"},{"key":"e_1_3_2_19_2","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1016\/0092-8674(95)90397-6","volume":"81","author":"Roelink H.","year":"1995","unstructured":"Roelink H., et al., Cell81, 445 (1995).","journal-title":"Cell"},{"key":"e_1_3_2_20_2","unstructured":"Induction of HNF3\u03b2 and Isl1 in midline explants was fully inhibited with 4.0 \u03bcM AY9944 1 \u03bcM triparanol and 1 \u03bcM cyclopamine. Intermediate doses of AY9944 (0.5 \u03bcM) triparanol (0.25 \u03bcM) and cyclopamine (0.25 \u03bcM) blocked induction of HNF3\u03b2 while permitting induction of Isl1."},{"key":"e_1_3_2_21_2","unstructured":"W. Gaffield and R. F. Keeler J. Nat. Toxins 5 25 (1996)."},{"key":"e_1_3_2_22_2","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1016\/S0022-2275(20)43173-6","volume":"31","author":"Gibson K. M.","year":"1990","unstructured":"HK293 cells, stably transfected with Shh with the Ecdysone-Inducible Mammalian Expression System (Invitrogen), were cultured to 30 to 40% confluency and then changed to culture medium with 10% delipidated serum [Gibson K. M., et al., J. Lipid Res. 31, 515 (1990)] and insulin-transferrin-sodium selenite media supplement (Sigma). After 24 hours, the cells were induced to express Shh with 1 \u03bcM muristerone A (Invitrogen), and AY9944, triparanol, jervine, cyclopamine, or tomatidine was added [see (15)]. After an additional 24 hours, cell lysates were analyzed with 12% SDS-polyacrylamide gels.","journal-title":"J. Lipid Res."},{"key":"e_1_3_2_23_2","unstructured":"Chick embryos were treated with \u223c10 \u03bcM jervine at stage 20 for 8 16 24 or 32 hours before fixation and stained with an antibody that recognizes Shh-N. No difference in the normal localization of Shh-N protein to the apical domain of floor plate cells or on the surfaces of commissural axons (16) was seen in treated embryos."},{"key":"e_1_3_2_24_2","unstructured":"Cleavage of the His 6 Hh-C protein was stimulated by 12 \u03bcM cholesterol but this cleavage was neither stimulated nor inhibited by the addition of jervine cyclopamine or tomatidine even at 350 \u03bcM."},{"key":"e_1_3_2_25_2","unstructured":"Other sterols that participated in the reaction with similar efficiency to cholesterol are \u03b2-sitosterol 5- androsten-3\u03b2-ol and 7\u03b2-hydroxycholesterol. Sterols that participated with reduced efficiency are coprastan-3-ol ergosterol 4\u03b2-hydroxycholesterol 19-hydroxycholesterol 20\u03b1-hydroxycholesterol 22( S )-hydroxycholesterol 22( R )-hydroxycholesterol and 25-hydroxycholesterol. Epicholesterol cholesterol acetate \u03b1-ecdysone 20-OH ecdysone 3-keto-5-cholestene and thiocholesterol (3(\u03b2)-thiol) were unable to participate in the reaction. The in vitro reaction thus requires an unhindered hydroxyl at the 3\u03b2 position on a tetracyclic sterol nucleus although neither the isooctyl side chain nor the number or positions of the double bonds in the sterol nucleus appear to affect autoprocessing critically."},{"key":"e_1_3_2_26_2","doi-asserted-by":"crossref","first-page":"661","DOI":"10.1016\/S0092-8674(00)81386-0","volume":"87","author":"Ericson J.","year":"1996","unstructured":"Ericson J., Morton S., Kawakami A., Roelink H., Jessell T. M., Cell87, 661 (1996).","journal-title":"Cell"},{"key":"e_1_3_2_27_2","first-page":"969","volume":"82","author":"Liem K. F.","year":"1995","unstructured":"Liem K. F., Tremml G., Roelink H., Jessell T. M., ibid82, 969 (1995).","journal-title":"ibid"},{"key":"e_1_3_2_28_2","unstructured":"Jervine (10 \u03bcM) cyclopamine (10 \u03bcM) and tomatidine (50 \u03bcM) all failed to inhibit formation of migratory HNK-1\u2013positive cells either from ventral neural plate explants cultured with BMP7 (100 ng\/ml) or from explanted dorsal neural plate with contiguous epidermal ectoderm (an endogenous source of BMP7)."},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.277.5329.1109"},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.277.5323.228"},{"key":"e_1_3_2_31_2","doi-asserted-by":"crossref","first-page":"1745","DOI":"10.1016\/S0021-9258(17)33712-2","volume":"251","author":"Bell J. J.","year":"1976","unstructured":"None of the explant responses to Shh-N were affected by addition of 25-hydroxycholesterol at 25 \u03bcM. 25-hydroxycholesterol is a potent inhibitor of HMG CoA reductase and at the concentrations used blocks de novo cholesterol synthesis in chick embryos and in cultured cell systems [S. C. Miller, G. Melnykovych, J. Lipid Res. 25, 991 (1984); Bell J. J., Sargeant T. E., Watson J. A., J. Biol. Chem. 251, 1745 (1976); (37)]. The addition of 25-hydroxycholesterol to explant cultures did not reverse the inhibitory effects of any of the teratogenic compounds.","journal-title":"J. Biol. Chem."},{"key":"e_1_3_2_32_2","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/S0387-7604(12)80118-5","volume":"13","author":"Yoshikawa H.","year":"1991","unstructured":"Yoshikawa H., Brain Dev.13, 115 (1991).","journal-title":"Brain Dev."},{"key":"e_1_3_2_33_2","doi-asserted-by":"crossref","first-page":"2627","DOI":"10.1074\/jbc.271.5.2627","volume":"271","author":"Metherall J. E.","year":"1996","unstructured":"Y. Lange, T. L. Steck, J. Biol. Chem. 269, 29371 (1994); Trends Cell Biol. 6, 205 (1996); Metherall J. E., Waugh K., Li H., J. Biol. Chem. 271, 2627 (1996); J. D. Butler,ibid. 267, 23797 (1992).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_2_34_2","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1016\/S0022-2275(20)37596-9","volume":"37","author":"Lange Y.","year":"1996","unstructured":"Lange Y., Duan H., Mazzone T., J. Lipid Res.37, 534 (1996).","journal-title":"J. Lipid Res."},{"key":"e_1_3_2_35_2","unstructured":"M. K. Cooper et al. unpublished data."},{"key":"e_1_3_2_36_2","unstructured":"The following class 2 sterol transport inhibitors at the given concentrations blocked the response of intermediate neural plate explants to 25 nM Shh-N without affecting signaling by BMP7: 0.25 \u03bcM U 18666A 50 \u03bcM chloroquine 75 \u03bcM imipramine and 20 \u03bcM progesterone."},{"key":"e_1_3_2_37_2","doi-asserted-by":"crossref","first-page":"987","DOI":"10.1242\/dev.120.4.987","volume":"120","author":"Capdevila J.","year":"1994","unstructured":"Capdevila J., Pariente P., Sampedro J., Alonso J. L., Guerrero I., Development120, 987 (1994); B. Wang, N. Fuse, J. A. Porter, P. A. Beachy, unpublished data.","journal-title":"Development"},{"key":"e_1_3_2_38_2","unstructured":"M. K. Cooper J. A. Porter K. E. Young P. A. Beachy data not shown."},{"key":"e_1_3_2_39_2","unstructured":"We thank R. I. Kelley Y. Lange D. Leahy and R. K. Mann for comments on the manuscript; S. Morton and T. Jessell for antibodies; W. Gaffield for plant alkaloids; Wyeth-Ayerst for AY9944; and Hoechst Marion Roussel for triparanol. M.K.C. is a Physician Postdoctoral Fellow and P.A.B. is an investigator of the Howard Hughes Medical Institute."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.280.5369.1603","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,13]],"date-time":"2024-01-13T04:09:01Z","timestamp":1705118941000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.280.5369.1603"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1998,6,5]]},"references-count":38,"journal-issue":{"issue":"5369","published-print":{"date-parts":[[1998,6,5]]}},"alternative-id":["10.1126\/science.280.5369.1603"],"URL":"https:\/\/doi.org\/10.1126\/science.280.5369.1603","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[1998,6,5]]}}}