메뉴 건너뛰기




Volumn 273, Issue 5276, 1996, Pages 784-788

Visualization of slow axonal transport in vivo

Author keywords

[No Author keywords available]

Indexed keywords

EPITOPE; NEUROFILAMENT PROTEIN; OLIGOMER; POLYMER;

EID: 0029778167     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.273.5276.784     Document Type: Article
Times cited : (78)

References (50)
  • 2
    • 0027358356 scopus 로고
    • B. Grafstein and D. S. Forman. Physiol. Rev. 60, 1167 (1980); N. Hirokawa, Curr. Opin. Neurobiol. 3, 724 (1993).
    • (1993) Curr. Opin. Neurobiol. , vol.3 , pp. 724
    • Hirokawa, N.1
  • 3
    • 0023024417 scopus 로고
    • Y. Komiya, M. Kurokawa, T. Tashiro, Biomed. Res. 7, 359 (1986); R. A. Nixon and K. B. Logvinenko, J. Cell Biol. 102, 647 (1986); T. Tashiro and Y. Komiya, J. Neurosci. 9, 760 (1989).
    • (1986) Biomed. Res. , vol.7 , pp. 359
    • Komiya, Y.1    Kurokawa, M.2    Tashiro, T.3
  • 4
    • 0022617701 scopus 로고
    • Y. Komiya, M. Kurokawa, T. Tashiro, Biomed. Res. 7, 359 (1986); R. A. Nixon and K. B. Logvinenko, J. Cell Biol. 102, 647 (1986); T. Tashiro and Y. Komiya, J. Neurosci. 9, 760 (1989).
    • (1986) J. Cell Biol. , vol.102 , pp. 647
    • Nixon, R.A.1    Logvinenko, K.B.2
  • 5
    • 0024504975 scopus 로고
    • Y. Komiya, M. Kurokawa, T. Tashiro, Biomed. Res. 7, 359 (1986); R. A. Nixon and K. B. Logvinenko, J. Cell Biol. 102, 647 (1986); T. Tashiro and Y. Komiya, J. Neurosci. 9, 760 (1989).
    • (1989) J. Neurosci. , vol.9 , pp. 760
    • Tashiro, T.1    Komiya, Y.2
  • 8
    • 0020474950 scopus 로고
    • R. J. Lasek, Philos. Trans. R. Soc. London B Biol. Sci. 299, 313 (1982); R. J. Lasek, J. A. Garner, S. T. Brady, J. Cell Biol. 99, 212s (1984); R. J. Lasek, J. Cell Sci. Suppl. 5, 161 (1986).
    • (1982) Philos. Trans. R. Soc. London B Biol. Sci. , vol.299 , pp. 313
    • Lasek, R.J.1
  • 9
    • 0021131071 scopus 로고
    • R. J. Lasek, Philos. Trans. R. Soc. London B Biol. Sci. 299, 313 (1982); R. J. Lasek, J. A. Garner, S. T. Brady, J. Cell Biol. 99, 212s (1984); R. J. Lasek, J. Cell Sci. Suppl. 5, 161 (1986).
    • (1984) J. Cell Biol. , vol.99
    • Lasek, R.J.1    Garner, J.A.2    Brady, S.T.3
  • 10
    • 0022849722 scopus 로고
    • R. J. Lasek, Philos. Trans. R. Soc. London B Biol. Sci. 299, 313 (1982); R. J. Lasek, J. A. Garner, S. T. Brady, J. Cell Biol. 99, 212s (1984); R. J. Lasek, J. Cell Sci. Suppl. 5, 161 (1986).
    • (1986) J. Cell Sci. Suppl. , vol.5 , pp. 161
    • Lasek, R.J.1
  • 11
    • 0025057456 scopus 로고
    • P. J. Hollenbeck, Nature 343, 408 (1990); D. W. Cleveland and P. N. Huffman, Cell 67, 453 (1991); H. C. Joshi and P.W. Baas, J. Cell Biol. 121, 1191 (1993).
    • (1990) Nature , vol.343 , pp. 408
    • Hollenbeck, P.J.1
  • 12
    • 0026041977 scopus 로고
    • P. J. Hollenbeck, Nature 343, 408 (1990); D. W. Cleveland and P. N. Huffman, Cell 67, 453 (1991); H. C. Joshi and P.W. Baas, J. Cell Biol. 121, 1191 (1993).
    • (1991) Cell , vol.67 , pp. 453
    • Cleveland, D.W.1    Huffman, P.N.2
  • 13
    • 0027221731 scopus 로고
    • P. J. Hollenbeck, Nature 343, 408 (1990); D. W. Cleveland and P. N. Huffman, Cell 67, 453 (1991); H. C. Joshi and P.W. Baas, J. Cell Biol. 121, 1191 (1993).
    • (1993) J. Cell Biol. , vol.121 , pp. 1191
    • Joshi, H.C.1    Baas, P.W.2
  • 14
    • 0027408530 scopus 로고    scopus 로고
    • S. Akli et al., Nature Genet. 3, 224 (1993); G. Bajocchi, S. H. Feldman, R. G. Crystal, A. Mastrangeli, ibid., p. 229; B. L. Davidson, E. D. Allen, K. F. Kozarsky, J. M. Wilson, B. J. Roessler, ibid., p. 219; G. Le Gal La Salle et al., Science 259, 988 (1993).
    • (1993) Nature Genet. , vol.3 , pp. 224
    • Akli, S.1
  • 15
    • 0027408530 scopus 로고    scopus 로고
    • S. Akli et al., Nature Genet. 3, 224 (1993); G. Bajocchi, S. H. Feldman, R. G. Crystal, A. Mastrangeli, ibid., p. 229; B. L. Davidson, E. D. Allen, K. F. Kozarsky, J. M. Wilson, B. J. Roessler, ibid., p. 219; G. Le Gal La Salle et al., Science 259, 988 (1993).
    • Nature Genet. , pp. 229
    • Bajocchi, G.1    Feldman, S.H.2    Crystal, R.G.3    Mastrangeli, A.4
  • 17
    • 0027457561 scopus 로고
    • S. Akli et al., Nature Genet. 3, 224 (1993); G. Bajocchi, S. H. Feldman, R. G. Crystal, A. Mastrangeli, ibid., p. 229; B. L. Davidson, E. D. Allen, K. F. Kozarsky, J. M. Wilson, B. J. Roessler, ibid., p. 219; G. Le Gal La Salle et al., Science 259, 988 (1993).
    • (1993) Science , vol.259 , pp. 988
    • Le Gal La Salle, G.1
  • 24
    • 0027220073 scopus 로고
    • G. Y. Ching and R. K. Liem, J. Cell Biol. 122, 1323 (1993); M. K. Lee, Z. Xu, P. C. Wong, D. W. Cleveland, ibid., p. 1337.
    • (1993) J. Cell Biol. , vol.122 , pp. 1323
    • Ching, G.Y.1    Liem, R.K.2
  • 27
    • 9444276433 scopus 로고    scopus 로고
    • note
    • The recombinant adenovirus vector AxCA-NT-NFM-myc was constructed by the COS-TPC method (26). Briefly, an expression cosmid cassette (pAxCA-NT-NFMmyc) was constructed by insertion of the expression unit (composed of the cytomegalovirus enhancer plus the chicken β-actin promoter, a complementary DNA coding sequence, and the rabbit β-globin polyadenylation signal sequence) into Swa I site of pAxw, which is a 42-kb cosmid containing a 31-kb adenovirus type 5 genome lacking E1A, E1B, and E3 genes. The expression cosmid cassette and the adenovirus DNA-terminal protein complex (Ad5 dlX DNA-TPC), predigested by Eco T221, were cotransfected into 293 cells [American Type Culture Collection (ATCC); CRL1573] by calcium phosphate precipitation. The recombinant viruses were subsequently propagated with 293 cells, and the viral solution was stored at -80°C. The titers of viral stocks were determined by plaque assay on 293 cells. For in vivo injection experiments, highly concentrated recombinant viral stocks were generated by the method of Kanegae et al. (15).
  • 28
    • 9444245705 scopus 로고    scopus 로고
    • note
    • 11 PFU/ml) recombinant adenovirus vector stock in phosphate buffered saline. Injected animals were killed between 3 and 7 days after surgery. Briefly, they were anesthetized with sodium pentobarbital and fixed by transcardiac perfusion with fresh 4% paraformaldehyde with (for immunoelectron microscopy) or without (for immunocytochemistry) 0.1% glutaraldehyde in 0.1 M phosphate buffer (pH 7.3). Immediately after perfusion, L4 dorsal roots, gangiia, and sciatic nerves were carefully excised. Consecutive 1-mm segments proceeding from the center of the DRG were removed and placed in the same fixative for an additional 2 hours at room temperature and for a further 12 hours at 4°C. The samples were then transferred to 10% sucrose in the same buffer and incubated for 2 hours at 4°C, after which they were cryoprotected with a graded series of sucrose concentrations, up to 60%, in the same phosphate buffer and then embedded In Tissue-Tek O.C.T. compound (Miles, Elkhart, IN) and frozen. For each segment, serial sections 8 to 10 μm thick were cut on a cryostat and collected. The sections were quenched (in 100 mM glycine) for 30 min at room temperature and then permeabilized and blocked (in 1% Triton X-100,0.1% saponin, and 5% skim milk) for 4 hours at room temperature; both solutions were prepared in 0.1 M phosphate buffer (pH 7.3). Subsequently, sections were incubated overnight at 4°C with a mAb recognizing the c-Myc epitope tag (Myc 1-9E10.2 hybridoma cells; ATCC; CRL1729), followed by a second overnight incubation at 4°C with either fluorescein isothiocyanate-labeled sheep anti-mouse immunoglobulin G (Amersham, Buckinghamshire, UK) for light microscopy, or with FluoroNanogold anti-mouse Fab' fluorescein (Nanoprobes, Stony Brook, NY) tor electron microscopy. All antibodies were diluted in the above-mentioned permeabilizing and blocking solution. Light microscopic analysis was performed by epifluorescent light microscopy (Axiophot; Zeiss, Oberkochen, Germany) or by confocal laser scanning microscopy (MRC-1000); Bio-Rad, Cambridge, MA). For analysis of confocal laser scanning microscope images, we used CoMOS software for image collection and qualitatively oriented image processing operations and MPL software for quantitatively oriented image processing operations. The sections labeled with FluoroNanogold anti-mouse Fab' fluorescein were detected by epifluoresence microscopy, fixed again in 1% glutaraldehyde with 0.2% tannic acid in phosphate buffer, rinsed with distilled water, and subsequently processed for silver enhancement in developer [30% gum arabic, 0.85% hydroquinone, and 0.11% silver nitrate in citrate buffer (pH 7.4)]; see Hayat (27) for detailed procedures. After enhancement, sections were stained with 1% uranyl acetate for 2 hours at room temperature. The samples were dehydrated in a graded series of ethanol concentrations and embedded in Epon 812. Ultrathin sections were cut on a conventional ultramicrotome, stained with uranyl acetate and lead citrate, and examined with a transmission electron microscope (1200EX; JEOL, Tokyo, Japan) at an accelerating voltage of 100 KV.
  • 32
    • 0024501230 scopus 로고
    • J. J. Blum and M. C. Reed, Cell Motil. Cytoskeleton 12, 53 (1989); R. A. Nixon, in The Neuronal Cytoskeleton, R. D. Burgoyne, Ed. (Wiley-Liss, New York, 1991).
    • (1989) Cell Motil. Cytoskeleton , vol.12 , pp. 53
    • Blum, J.J.1    Reed, M.C.2
  • 33
    • 0004079567 scopus 로고
    • R. D. Burgoyne, Ed. Wiley-Liss, New York
    • J. J. Blum and M. C. Reed, Cell Motil. Cytoskeleton 12, 53 (1989); R. A. Nixon, in The Neuronal Cytoskeleton, R. D. Burgoyne, Ed. (Wiley-Liss, New York, 1991).
    • (1991) The Neuronal Cytoskeleton
    • Nixon, R.A.1
  • 35
    • 9444298294 scopus 로고    scopus 로고
    • note
    • 125l-labeled protein A (Amersham) and analyzed with an image analyzer (BAS 2000; Fuji, Tokyo, Japan).
  • 38
    • 0027967986 scopus 로고
    • J. F. Engelhardt, X. Ye, B. Doranz, J. M. Wilson. Proc. Natl. Acad. Sci. U.S.A. 91, 6196 (1994); B. R. Grubb et al., Nature 371, 802 (1994); Y. Yang et al., Nature Genet. 7, 362 (1994); Y. Yang et al., Proc. Natl. Acad. Sci. U.S.A. 91, 4407 (1994).
    • (1994) Nature , vol.371 , pp. 802
    • Grubb, B.R.1
  • 39
    • 0028229147 scopus 로고
    • J. F. Engelhardt, X. Ye, B. Doranz, J. M. Wilson. Proc. Natl. Acad. Sci. U.S.A. 91, 6196 (1994); B. R. Grubb et al., Nature 371, 802 (1994); Y. Yang et al., Nature Genet. 7, 362 (1994); Y. Yang et al., Proc. Natl. Acad. Sci. U.S.A. 91, 4407 (1994).
    • (1994) Nature Genet. , vol.7 , pp. 362
    • Yang, Y.1
  • 40
    • 0028328261 scopus 로고
    • J. F. Engelhardt, X. Ye, B. Doranz, J. M. Wilson. Proc. Natl. Acad. Sci. U.S.A. 91, 6196 (1994); B. R. Grubb et al., Nature 371, 802 (1994); Y. Yang et al., Nature Genet. 7, 362 (1994); Y. Yang et al., Proc. Natl. Acad. Sci. U.S.A. 91, 4407 (1994).
    • (1994) Proc. Natl. Acad. Sci. U.S.A. , vol.91 , pp. 4407
    • Yang, Y.1
  • 41
    • 9444257935 scopus 로고    scopus 로고
    • unpublished data
    • J.-P. Julien et al., unpublished data.
    • Julien, J.-P.1
  • 46
    • 0021874883 scopus 로고
    • I. Saito, Y. Oya, K. Yamamoto, T. Yuasa, H. Shimojo, J. Virol. 54, 711 (1985); Y. Kanegae et al. Nucleic Acids Res. 23, 3816 (1995); S. Miyake et al., Proc. Natl. Acad. Sci, U.S.A. 93, 1320 (1996).
    • (1985) J. Virol. , vol.54 , pp. 711
    • Saito, I.1    Oya, Y.2    Yamamoto, K.3    Yuasa, T.4    Shimojo, H.5
  • 47
    • 0028799961 scopus 로고
    • I. Saito, Y. Oya, K. Yamamoto, T. Yuasa, H. Shimojo, J. Virol. 54, 711 (1985); Y. Kanegae et al. Nucleic Acids Res. 23, 3816 (1995); S. Miyake et al., Proc. Natl. Acad. Sci, U.S.A. 93, 1320 (1996).
    • (1995) Nucleic Acids Res. , vol.23 , pp. 3816
    • Kanegae, Y.1
  • 48
    • 0030070724 scopus 로고    scopus 로고
    • I. Saito, Y. Oya, K. Yamamoto, T. Yuasa, H. Shimojo, J. Virol. 54, 711 (1985); Y. Kanegae et al. Nucleic Acids Res. 23, 3816 (1995); S. Miyake et al., Proc. Natl. Acad. Sci, U.S.A. 93, 1320 (1996).
    • (1996) Proc. Natl. Acad. Sci, U.S.A. , vol.93 , pp. 1320
    • Miyake, S.1
  • 50
    • 9444220632 scopus 로고    scopus 로고
    • note
    • We thank I. Saito and Y. Kanegae (Institute of Medical Science, University of Tokyo) and J. Miyazaki (University of Tokyo School of Medicine) for supplying us with their recombinant adenovirus vector system and the CAG promoter, respectively. Their technical advice was essential for the successful construction of our vector. We also thank Y. Komiya and T. Tashiro (Gunma University School of Medicinel for their technical assistance and advice on the microinjection into dorsal root ganglia in vivo; S. Ijuin (Nippon Bio-Rad Laboratories) for his technical assistance in confocal laser scanning microscopy; H. Yorifuji (National Defense Medical College) for his guidance in electron microscopic procedures; J.-P. Julien (Montreal General Hospital Research Institute, Centre for Research in Neuroscience, McGill University) and R. Nixon (McLean Hospital, Harvard Medical School) for helpful comments on the manuscript and useful discussions; Y. Kobayashi (Kyorin University School of Medicine) and T. Funakoshi, S. Kondo, Y. Okada, and other members of the Hirokawa lab for stimulating discussions and invaluable advice throughout; and Y. Kawasaki, H. Sato, H. Fukuda, and Y. Sugaya for their technical and secretarial assistance. Supported by a Special Grant-in-Aid for Scientific Research from the Japan Ministry of Educatton, Science and Culture; a grant from the Institute of Physical and Chemical Research (RIKEN) to N.H.; a Grant-in-Aid for Scientific Research from the Japan Ministry of Education, Science and Culture to S.T.; and grants from the Medical Research Council of Canada and Muscular Dystrophy Association of Canada to A.C.P.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.