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Volumn 275, Issue 5306, 1997, Pages 1652-1654

A member of the frizzled protein family mediating axis induction by Wnt- 5A

Author keywords

[No Author keywords available]

Indexed keywords

ARTICLE; EMBRYO; EMBRYO AXIS; EMBRYO DEVELOPMENT; NONHUMAN; PRIORITY JOURNAL; RECEPTOR BINDING; SIGNAL TRANSDUCTION; XENOPUS LAEVIS;

EID: 0030941498     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.275.5306.1652     Document Type: Article
Times cited : (423)

References (36)
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    • A. P. McMahon and R. T. Moon, Cell 58, 1075 (1989); W. C. Smith and R. M. Harland, ibid. 67, 753 (1991); S. Sokol, J. L. Christian, R. T. Moon, D. A. Melton, ibid., p. 741; A. Chakrabarti, G. Matthews, A. Colman, L. Dale, Development 115, 355 (1992); S. L. Wolda, C. J. Moody, R. T. Moon, Dev. Biol. 155, 46 (1993).
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    • note
    • All fz cDNAs were subcloned in the pRK5 vector [C. Gorman, D. R. Gies, G. McCray, DNA Prot. Eng. Techn. 2, 3 (1990)] with an optimal Kozak consensus sequence for translation at the initiator ATG (CCAC-CATG, preceded by different restriction sites for subcloning) and with different lengths of 3′ untranslated regions. The details of each construct are available upon request. Xwnt-5A was in pSP64 vector as described (8). All RNAs for injection were synthesized as capped transcripts in vitro with SP6 RNA polymerase (Ambion Megascript). Unless otherwise specified, RNAs were injected into the two cells near the equatorial midline region at the four-cell stage. Embryo preparation and staging were performed as described (24).
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    • Percentages of axis duplication were obtained by dividing the total number of embryos with a duplicated axis by the total number of embryos, scored from two to six independent experiments. Extensive axis duplication is defined by the presence of the cement gland and at least one eye in the duplicated axis.
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    • note
    • Murine wnt-5 cDNA (14) was cloned in pCS2+ vector [D. L. Turner and H. Weintraub, Genes Dev. 8, 1434 (1994)] as the Eco RI and Xba I fragment, which contains about 60 bp 5′ and 360 bp 3′ untranslated regions.
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    • note
    • Some variability exists in the completeness of duplicated axes or axial rescue in UV-treated embryos among different batches of embryos. One possibility is that the coinjection of Xwnt-5A and hfz5 RNAs inevitably causes more variable results than does injection of a single RNA species, because the two proteins derived from the injected RNAs must be translated, processed, and localized properly to function together. In injection of a single RNA, such as one for Xwnt-8, the variables are only associated with the Xwnt-8 protein, because one or more receptors are provided endogenously. Another possibility is that Xwnt-5A inhibits gastrulation movements in gastrula stages (8); this later function of Xwnt-5A might affect the completeness of the axis to a variable degree.
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    • X. He et al., data not shown
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    • Embryo preparation, staging, UV irradiation, fixation and sectioning, and whole-mount in situ hybridization were performed as described (24).
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    • note
    • hfz5N was generated by introduction of a stop codon just before the first putative transmembrane helix (changing the preceding amino acids 237 and 238 from phenylalanine and tryptophan to threonine and arginine, respectively). The corresponding DNA fragment was subcloned in pCS2+.
  • 36
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    • note
    • The authors are grateful to R. Moon for Xwnt-5A plasmid. We thank B. Huang for technical help and B. Williams, J. Debnath, P. Schwartzberg, and M. Rebagliati for discussion and comments on the manuscript. X.H. is supported by the U.S. Army Breast Cancer Research Program. J.N. is an investigator of the Howard Hughes Medical Institute.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.