Altered neural cell fates and medulloblastoma in mouse patched mutants

Science

Volumn 277, Issue 5329, 1997, Pages 1109-1113

  Goodrich, Lisa V ^a   Milenković, Ljiljana ^a   Higgins, Kay M ^a   Scott, Matthew P ^a  

^a STANFORD UNIVERSITY SCHOOL OF MEDICINE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ANIMAL CELL; ARTICLE; BINDING AFFINITY; ENDODERM; GENE ACTIVATION; GENE TARGETING; MEDULLOBLASTOMA; MOUSE; NERVE CELL; NONHUMAN; PRIORITY JOURNAL; REGULATOR GENE; TARGET CELL; TUMOR SUPPRESSOR GENE;

ABNORMALITIES, MULTIPLE; ANIMALS; BODY PATTERNING; CELL LINEAGE; CENTRAL NERVOUS SYSTEM; CEREBELLAR NEOPLASMS; ECTODERM; ENDODERM; GENE EXPRESSION REGULATION, DEVELOPMENTAL; GENES, TUMOR SUPPRESSOR; HETEROZYGOTE; HOMOZYGOTE; INTRACELLULAR SIGNALING PEPTIDES AND PROTEINS; MEDULLOBLASTOMA; MEMBRANE PROTEINS; MESODERM; MICE; MICE, INBRED C57BL; MUTATION; ONCOGENE PROTEINS; RECEPTORS, CELL SURFACE; TRANS-ACTIVATORS; TRANSCRIPTION FACTORS;

ANIMALIA; ERINACEIDAE;

EID: 0030866154     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.277.5329.1109     Document Type: Article

References (47)

1. H. Hahn et al., Cell 85, 841 (1996); R. L. Johnson et al., Science 272, 1668 (1996).
2. H. Hahn et al., Cell 85, 841 (1996); R. L. Johnson et al., Science 272, 1668 (1996).
3. R. J. Gorlin, Medicine 66, 98 (1987).
4. M. Hammerschmidt, A. Brook, A. P. McMahon, Trends Genet. 13, 14 (1997).
5. P. W. Ingham, A. M. Taylor, Y. Nakano, Nature 353, 184 (1991).
6. P. W. Ingham, Curr. Biol. 4, 347 (1994).
7. L. V. Goodrich, R. L. Johnson, L. Milenkovic, J. McMahon, M. P. Scott, Genes Dev. 10, 301 (1996); V. Marigo, M. P. Scott, R. L. Johnson, L. Goodrich, C. J. Tabin, Development 122, 1225 (1996); J.-P. Concordet et al., ibid., p. 2835.
8. L. V. Goodrich, R. L. Johnson, L. Milenkovic, J. McMahon, M. P. Scott, Genes Dev. 10, 301 (1996); V. Marigo, M. P. Scott, R. L. Johnson, L. Goodrich, C. J. Tabin, Development 122, 1225 (1996); J.-P. Concordet et al., ibid., p. 2835.
9. L. V. Goodrich, R. L. Johnson, L. Milenkovic, J. McMahon, M. P. Scott, Genes Dev. 10, 301 (1996); V. Marigo, M. P. Scott, R. L. Johnson, L. Goodrich, C. J. Tabin, Development 122, 1225 (1996); J.-P. Concordet et al., ibid., p. 2835.
10. V. Marigo et al., Nature 384, 176 (1996); D. M. Stone et al., ibid., p. 129.
11. V. Marigo et al., Nature 384, 176 (1996); D. M. Stone et al., ibid., p. 129.
12. A. E. Oro et al., Science 276, 817 (1997).
13. D. Lacombe, J. F. Chateil, D. Fontan, J. Battin, Genet. Couns. 1, 273 (1990).
14. B. Novakovic, Med. Pediatr. Oncol. 23, 480 (1994).
15. C. Raffel et al., Cancer Res. 57, 842 (1997); J. Xie et al., ibid., p. 2369.
16. C. Raffel et al., Cancer Res. 57, 842 (1997); J. Xie et al., ibid., p. 2369.
17. K. M. Fung and J. Q. Trojanowski, J. Neuropathol. Exp. Neurol. 54, 285 (1995); L. B. Rorke, Am. J. Pathol. 144, 444 (1994).
18. K. M. Fung and J. Q. Trojanowski, J. Neuropathol. Exp. Neurol. 54, 285 (1995); L. B. Rorke, Am. J. Pathol. 144, 444 (1994).
19. DNA from the ptc genomic locus was isolated from a 129SV genomic phage library (Stratagene). Exons 1 through 15 of human PTC (1) were mapped by polymerase chain reaction (PCR) and sequencing. The 3′ arm of homology was a 3.5-kb Eco RI-Bam HI fragment from the second intron that gained a Bam HI site from pBSII (Stratagene) and was cloned into the Bam HI site of pPNT [V. L. Tybulewicz, C. E. Crawford, P. K. Jackson, R. T. Bronson, R. C. Mulligan, Cell 65, 1153 (1991)]. A cassette containing the gene for nuclear localized β-galactosidase (lacZ) followed by the mP1 intron and poladenylate tail was excised from pNLacF [E. H. Mercer, G. W. Hoyle, R. P. Kapur, R. L. Brinster, R. D. Palmiter, Neuron 7, 703 (1991)] and cloned into the Xho I site of pPNT by using Xho I and Sal I linkers. The 5′ arm of homology was a 6.5-kb Xho I-Nru I fragment that was cloned into the Xho I site upstream of lacZ with a Sal I linker. The Nru I site is in the first ptc exon. The resulting plasmid, KO1, was linearized with Xho I and electroporated into RI embryonic stem cells that were subjected to double selection and analyzed by Southern (DNA) blot [A. L. Joyner, Gene Targeting: A Practical Approach (Oxford Univ. Press, New York, 1993), pp. 33-61]. Targeted clones were expanded and used for injection into C57BI/6 blastocysts [B. Hogan, R. Beddington, F. Costantini, E. Lacy, Manipulating the Mouse Embryo: A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1994), pp. 196-204].
20. DNA from the ptc genomic locus was isolated from a 129SV genomic phage library (Stratagene). Exons 1 through 15 of human PTC (1) were mapped by polymerase chain reaction (PCR) and sequencing. The 3′ arm of homology was a 3.5-kb Eco RI-Bam HI fragment from the second intron that gained a Bam HI site from pBSII (Stratagene) and was cloned into the Bam HI site of pPNT [V. L. Tybulewicz, C. E. Crawford, P. K. Jackson, R. T. Bronson, R. C. Mulligan, Cell 65, 1153 (1991)]. A cassette containing the gene for nuclear localized β-galactosidase (lacZ) followed by the mP1 intron and poladenylate tail was excised from pNLacF [E. H. Mercer, G. W. Hoyle, R. P. Kapur, R. L. Brinster, R. D. Palmiter, Neuron 7, 703 (1991)] and cloned into the Xho I site of pPNT by using Xho I and Sal I linkers. The 5′ arm of homology was a 6.5-kb Xho I-Nru I fragment that was cloned into the Xho I site upstream of lacZ with a Sal I linker. The Nru I site is in the first ptc exon. The resulting plasmid, KO1, was linearized with Xho I and electroporated into RI embryonic stem cells that were subjected to double selection and analyzed by Southern (DNA) blot [A. L. Joyner, Gene Targeting: A Practical Approach (Oxford Univ. Press, New York, 1993), pp. 33-61]. Targeted clones were expanded and used for injection into C57BI/6 blastocysts [B. Hogan, R. Beddington, F. Costantini, E. Lacy, Manipulating the Mouse Embryo: A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1994), pp. 196-204].
21. DNA from the ptc genomic locus was isolated from a 129SV genomic phage library (Stratagene). Exons 1 through 15 of human PTC (1) were mapped by polymerase chain reaction (PCR) and sequencing. The 3′ arm of homology was a 3.5-kb Eco RI-Bam HI fragment from the second intron that gained a Bam HI site from pBSII (Stratagene) and was cloned into the Bam HI site of pPNT [V. L. Tybulewicz, C. E. Crawford, P. K. Jackson, R. T. Bronson, R. C. Mulligan, Cell 65, 1153 (1991)]. A cassette containing the gene for nuclear localized β-galactosidase (lacZ) followed by the mP1 intron and poladenylate tail was excised from pNLacF [E. H. Mercer, G. W. Hoyle, R. P. Kapur, R. L. Brinster, R. D. Palmiter, Neuron 7, 703 (1991)] and cloned into the Xho I site of pPNT by using Xho I and Sal I linkers. The 5′ arm of homology was a 6.5-kb Xho I-Nru I fragment that was cloned into the Xho I site upstream of lacZ with a Sal I linker. The Nru I site is in the first ptc exon. The resulting plasmid, KO1, was linearized with Xho I and electroporated into RI embryonic stem cells that were subjected to double selection and analyzed by Southern (DNA) blot [A. L. Joyner, Gene Targeting: A Practical Approach (Oxford Univ. Press, New York, 1993), pp. 33-61]. Targeted clones were expanded and used for injection into C57BI/6 blastocysts [B. Hogan, R. Beddington, F. Costantini, E. Lacy, Manipulating the Mouse Embryo: A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1994), pp. 196-204].
22. DNA from the ptc genomic locus was isolated from a 129SV genomic phage library (Stratagene). Exons 1 through 15 of human PTC (1) were mapped by polymerase chain reaction (PCR) and sequencing. The 3′ arm of homology was a 3.5-kb Eco RI-Bam HI fragment from the second intron that gained a Bam HI site from pBSII (Stratagene) and was cloned into the Bam HI site of pPNT [V. L. Tybulewicz, C. E. Crawford, P. K. Jackson, R. T. Bronson, R. C. Mulligan, Cell 65, 1153 (1991)]. A cassette containing the gene for nuclear localized β-galactosidase (lacZ) followed by the mP1 intron and poladenylate tail was excised from pNLacF [E. H. Mercer, G. W. Hoyle, R. P. Kapur, R. L. Brinster, R. D. Palmiter, Neuron 7, 703 (1991)] and cloned into the Xho I site of pPNT by using Xho I and Sal I linkers. The 5′ arm of homology was a 6.5-kb Xho I-Nru I fragment that was cloned into the Xho I site upstream of lacZ with a Sal I linker. The Nru I site is in the first ptc exon. The resulting plasmid, KO1, was linearized with Xho I and electroporated into RI embryonic stem cells that were subjected to double selection and analyzed by Southern (DNA) blot [A. L. Joyner, Gene Targeting: A Practical Approach (Oxford Univ. Press, New York, 1993), pp. 33-61]. Targeted clones were expanded and used for injection into C57BI/6 blastocysts [B. Hogan, R. Beddington, F. Costantini, E. Lacy, Manipulating the Mouse Embryo: A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1994), pp. 196-204].
23. Y. Chen and G. Struhl, Cell 87, 553 (1996).
24. C. K. Motzny and R. Holmgren, Mech. Dev. 52, 137 (1995); M. Domínguez, M. Brunner, E. Hafen, K. Basler, Science 272, 1621 (1996); J. Hepker, Q. T. Wang, C. K. Motzny, R. Holmgren, T. V. Orenic, Development 124, 549 (1997); P. Aza-Blanc, F.-A. Ramirez-Weber, M.-P. Laget, C. Schwartz, T. B. Kornberg, Cell 89, 1043 (1997).
25. C. K. Motzny and R. Holmgren, Mech. Dev. 52, 137 (1995); M. Domínguez, M. Brunner, E. Hafen, K. Basler, Science 272, 1621 (1996); J. Hepker, Q. T. Wang, C. K. Motzny, R. Holmgren, T. V. Orenic, Development 124, 549 (1997); P. Aza-Blanc, F.-A. Ramirez-Weber, M.-P. Laget, C. Schwartz, T. B. Kornberg, Cell 89, 1043 (1997).
26. C. K. Motzny and R. Holmgren, Mech. Dev. 52, 137 (1995); M. Domínguez, M. Brunner, E. Hafen, K. Basler, Science 272, 1621 (1996); J. Hepker, Q. T. Wang, C. K. Motzny, R. Holmgren, T. V. Orenic, Development 124, 549 (1997); P. Aza-Blanc, F.-A. Ramirez-Weber, M.-P. Laget, C. Schwartz, T. B. Kornberg, Cell 89, 1043 (1997).
27. C. K. Motzny and R. Holmgren, Mech. Dev. 52, 137 (1995); M. Domínguez, M. Brunner, E. Hafen, K. Basler, Science 272, 1621 (1996); J. Hepker, Q. T. Wang, C. K. Motzny, R. Holmgren, T. V. Orenic, Development 124, 549 (1997); P. Aza-Blanc, F.-A. Ramirez-Weber, M.-P. Laget, C. Schwartz, T. B. Kornberg, Cell 89, 1043 (1997).
28. V. Marigo et al., Dev. Biol. 180, 273 (1996).
29. D. J. Roberts et al., Development 121, 3163 (1995).
30. Y. Echelard et al., Cell 75, 1417 (1993); H. Roelink et al., ibid. 76, 761 (1994); H. Roelink et al., ibid. 81, 445 (1995).
31. Y. Echelard et al., Cell 75, 1417 (1993); H. Roelink et al., ibid. 76, 761 (1994); H. Roelink et al., ibid. 81, 445 (1995).
32. Y. Echelard et al., Cell 75, 1417 (1993); H. Roelink et al., ibid. 76, 761 (1994); H. Roelink et al., ibid. 81, 445 (1995).
33. C. Chiang et al., Nature 383, 407 (1996).
34. C. Walther and P. Gruss, Development 113, 1435 (1991).
35. M. E. Dickinson et al., ibid. 121, 2099 (1995); K. Liem Jr., G. Tremml, H. Roelink, T. M. Jessell, Cell 82, 969 (1995).
36. M. E. Dickinson et al., ibid. 121, 2099 (1995); K. Liem Jr., G. Tremml, H. Roelink, T. M. Jessell, Cell 82, 969 (1995).
37. H. U. Wang and D. J. Anderson, Neuron 18, 383 (1997).
38. K. Shimamura et al., Development 121, 3923 (1995).
39. D. G. Wilkinson et al., Nature 341, 405 (1989).
40. J. P. Provias and L. E. Becker, J. Neurooncol. 29, 35 (1996).
41. For immunostaining, two tumors were fixed and embedded in paraffin. Tissue sections (4 μm thick) were cleared and dehydrated, treated with 3% hydrogen peroxide, and then treated with a dilution of 1:10 normal rabbit serum (Vector Laboratories). Antibody to synaptophysin (Boehringer-Mannheim) was used at a dilution of 1:5 and antibody to neurofilament protein (Dako) at 1:50. Antibody binding was visualized with a peroxidase Vectastain Elite ABC kit (Vector Laboratories). Nuclei were counterstained with hematoxylin. As with anti-synaptophysin staining, anti-neurofilament staining appeared in processes of the tumor cells.
42. L. Stevenson and F. Echlin, Arch. Neurol. Psychiatry 31, 93 (1934); M. E. Kadin, L. J. Rubinstein, J. S. Nelson, J. Neuropathol. Exp. Neurol. 29, 583 (1970).
43. L. Stevenson and F. Echlin, Arch. Neurol. Psychiatry 31, 93 (1934); M. E. Kadin, L. J. Rubinstein, J. S. Nelson, J. Neuropathol. Exp. Neurol. 29, 583 (1970).
44. Embryos were stained with X-Gal (5-bromo-4-chloro-3-indoxyl-β-D-galactopyranoside) [C. Bonnerot and J. Nicolas, in Guide to Techniques in Mouse Development, P. M. Wassarman and M. L. DePamphilis, Eds. (Academic Press, New York, 1993), p. 463] and then sectioned. For X-Gal staining of tumors in heterozygotes, cerebellums were dissected, cut into four pieces along the sagittal plane, fixed briefly in 4% paraformaldehyde, and then stained.
45. D. G. Wilkinson, in In situ Hybridization: A Practical Approach, D. G. Wilkinson, Ed. (Oxford Univ. Press, Oxford, 1992), pp. 75-83.
46. M. C. Green, Ohio J. Sci. 52, 31 (1952).
47. We thank D. Kingsley, S. McConnell, and G. Barsh for help with this project; D. Horoupian for advice on medulloblastoma histology; D. Jung and N. Jiang for technical assistance; A. Oro for helpful discussions; K. Wharton and L. Havton for advice and tissue samples; D. Anderson, P. Gruss, R. Krumlauf, A. McMahon, A. Nagy, J. Rossant, J. Rubinstein, A. Simeone, and D. Wilkinson for research materials; and the Protein and Nucleic Acid facility at Stanford for DNA help. L.V.G. is supported by an Howard Hughes Medical Institute predoctoral fellowship. This research was supported by the Howard Hughes Medical Institute. Mouse work was done according to Animal Use Guidelines at Stanford University.