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Volumn 288, Issue 5468, 2000, Pages 1047-1051

N-cadherin, a cell adhesion molecule involved in establishment of embryonic left-right asymmetry

Author keywords

[No Author keywords available]

Indexed keywords

NERVE CELL ADHESION MOLECULE;

EID: 0040784424     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.288.5468.1047     Document Type: Article
Times cited : (132)

References (42)
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    • In situ hybridization of antisense RNA probes was performed as described [A. K. Knecht, P. J. Good, I. B. Dawid, R. M. Harland, Development 121, 1927 (1995); D. Henrique et al., Nature 375, 787 (1995)]. The full-length chicken N-cadherin (gift of M. Takeichi) was digested with Pst I, and a 461-base-pair fragment corresponding to the extracellular domain was cloned into pBS II SK+ and used to detect N-cadherin expression. Other probes were used as recommended and provided as follows: Nodal (M. R. Kuehn), Snail (A. Nieto), Lefty and Pitx2 (J. C. Izpizua-Belmonte), Shh and FGF8 (C. Tabin), and ActRIIa (C. Stern).
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    • White Leghorn chicken embryos were staged according to Hamburger and Hamilton (30). Antibodies were used as follows: 1 μl of saturated hybridoma supernatant of monoclonal antibody NCD-2, anti-N-cadherin, or control antibody IIH6C4 (chosen as a control because it recognizes an extracellular epitope present in both the primitive streak and the node of stage 4 to 6 chicken embryos) was applied from above to the ventral side of the node in embryos prepared for New culture according to [D. A. T. New, J. Embryol. Exp. Morphol. 3, 320 (1955)]. Affi-Gel beads (Bio-Rad) were rinsed three times in large volumes of phosphate-buffered saline (PBS) and incubated for 1 hour at room temperature in 0.1% BSA in PBS solution (control) or with activin (R&D Systems; 10 units/μl) or Shh (from E. Turner; 4 μg/μl) and rinsed briefly before implantation between the epiblast and endoderm. These dosages of activin and Shh were sufficient to randomize the direction of heart looping in our hands.
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    • note
    • Supported by U.S. Public Health Service grant HD 15527 (M.B.-F.). American Heart Association 1140-FI1 (M.I.G-C.), and the Summer Undergraduate Research Fellowship, Caltech (E.V.). We thank C. Baker, A. Groves, C. LaBonne, and B. E. Murray for helpful discussions and comments and R. Velasco for help with sectioning.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.