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JOURNAL ARTICLES
Defects in heart and lung development in compound heterozygotes for two different targeted mutations at the N-myc locus
C.B. Moens, B.R. Stanton, L.F. Parada, J. Rossant
Development 1993 119: 485-499;

Summary

Two types of mutant allele, one leaky and one null, have been generated by gene targeting at the N-myc locus in embryonic stem cells and the phenotypes of mice homozygous for these mutations have been described. These mutations have shown that N-myc has a number of functions during development, including a role in branching morphogenesis in the lung, which manifests itself at birth in mice homozygous for the leaky allele, and roles in the development of the mesonephric tubules, the neuroepithelium, the sensory ganglia, the gut and the heart, which become evident at midgestation in embryos homozygous for the null allele. In an attempt to define roles for N-myc at other stages of development, we have combined the two types of N-myc mutant allele in a compound heterozygote that as a result contains approximately 15% of normal levels of N-Myc protein. Compound heterozygotes died during gestation at a time intermediate to the times of death of embryos homozygous for either mutation individually, and their death appeared to result from cardiac failure stemming from hypoplasia of the compact subepicardial layer of the myocardium. Investigation of the expression pattern of N-myc and various markers of differentiation in wild-type and compound heterozygote mutant hearts has suggested that N-myc may function in maintaining the proliferation and/or preventing the differentiation of compact layer myocytes. This study illustrates the importance of generating different mutations at a given locus to elucidate fully the function of a particular gene during development.

REFERENCES

    1. Adams J. M.,
    2. Harris A. W.,
    3. Pinkert C. A.,
    4. Corcoran L. M.,
    5. Alexander W. S.,
    6. Cory S.,
    7. Palmiter R. D.,
    8. Brinster R. L.
    (1985). The c-myc oncogene driven by immunoglobulin enhancers induces lymphoid malignancy in transgenic mice. Nature 318, 533–538
    OpenUrlCrossRefPubMedWeb of Science
    1. Alex R.,
    2. Soezeri O.,
    3. Meyer S.,
    4. Dildrop R.
    (1992). Determination of the DNA sequence recognized by the bHLH-zip domain of the N-Myc protein. Nucleic Acids Res 20, 2257–2263
    OpenUrlAbstract/FREE Full Text
    1. Amati B.,
    2. Dalton S.,
    3. Brooks M. W.,
    4. Littlewood T. D.,
    5. Evan G. I.,
    6. Land H.
    (1992). Transcriptional activation by the human c-Myc oncoprotein in yeast requires interaction with Max. Nature 359, 423–426
    OpenUrlCrossRefPubMed
    1. Amati B.,
    2. Brooks M. W.,
    3. Levy N.,
    4. Littlewood T. D.,
    5. Evan G. I.,
    6. Land H.
    (1993). Oncogenic activity of the c-Myc protein requires dimerization with Max. Cell 72, 233–245
    OpenUrlCrossRefPubMedWeb of Science
    1. Ayer D. E.,
    2. Kretzner L.,
    3. Eisenman R. N.
    (1993). Mad: a heterodimeric partner for Max that antagonizes Myc transcriptional activity. Cell 72, 211–222
    OpenUrlCrossRefPubMedWeb of Science
    1. Barrett J.,
    2. Birrer M. J.,
    3. Kato G. J.,
    4. Dosaka-Akita H.,
    5. Dang C. V.
    (1992). Activation domains of L-Myc and c-Myc determine their transforming potencies in rat embryo cells. Mol. Cell Biol 12, 3130–3137
    OpenUrlAbstract/FREE Full Text
    1. Blackwell T. K.,
    2. Kretzner L.,
    3. Blackwood E. M.,
    4. Eisenman R. N.,
    5. Weintraub H.
    (1990). Sequence-specific DNA binding by the c-Myc protein. Science 250, 1149–1151
    OpenUrlAbstract/FREE Full Text
    1. Blackwood E. M.,
    2. Luescher B.,
    3. Eisenman R. N.
    (1992). Myc and Max associate in vivo. Genes Dev 6, 71–80
    OpenUrlAbstract/FREE Full Text
    1. Blackwood E. M.,
    2. Eisenman R. N.
    (1991). Max: a helix-loop-helix zipper protein that forms a sequence-specific DNA-binding complex with Myc. Science 251, 1211–1217
    OpenUrlAbstract/FREE Full Text
    1. Bossone S. A.,
    2. Asselin C.,
    3. Patel A. J.,
    4. Marcu K. B.
    (1992). Maz, a zinc finger protein, binds to c-MYC and C2 gene sequences regulating transcriptional initiation and termination. Proc. Natl. Acad. Sci. USA 89, 7452–7456
    OpenUrlAbstract/FREE Full Text
    1. Challice C. E.,
    2. Viragh S.
    (1973). The architectural development of the early mammalian heart. Tissue Cell 6, 447–462
    OpenUrlCrossRef
    1. Charron J.,
    2. Malynn B. A.,
    3. Robertson E. J.,
    4. Goff S. P.,
    5. Alt F. W.
    (1990). High-frequency disruption of the N-myc gene in embryonic stem and pre-B cell lines by homologous recombination. Mol. Cell Biol 10, 1799–1804
    OpenUrlAbstract/FREE Full Text
    1. Charron J.,
    2. Malynn B. A.,
    3. Fisher P.,
    4. Stewart V.,
    5. Jeannotte L.,
    6. Goff S. P.,
    7. Robertson E. J.,
    8. Alt F. W.
    (1992). Embryonic lethality in mice homozygous for a targeted disruption of the N-myc gene. Genes Dev 6, 2248–2257
    OpenUrlAbstract/FREE Full Text
    1. Davis A. C.,
    2. Wims M.,
    3. Spotts G. D.,
    4. Hann S. R.,
    5. Bradley A.
    (1993). A null c- myc mutation causes lethality before 10.5 days of gestation in homozygotes and reduced fertility in heterozygous female mice. Genes Dev 7, 671–682
    OpenUrlAbstract/FREE Full Text
    1. DePinho R. A.,
    2. Legouy E.,
    3. Feldman L. B.,
    4. Kohl N. E.,
    5. Yancopoulos G. D.,
    6. Alt F. W.
    (1986). Structure and expression of the murine N-myc gene. Proc. Natl. Acad. Sci. USA 83, 1827–1831
    OpenUrlAbstract/FREE Full Text
    1. DePinho R. A.,
    2. Schreiber-Agus N.,
    3. Alt F. W.
    (1991). myc family oncogenes in the development of normal and neoplastic cells. Adv. Cancer Res 57, 1–46
    OpenUrlCrossRefPubMedWeb of Science
    1. Dildrop R.,
    2. Ma A.,
    3. Zimmerman K.,
    4. Hsu E.,
    5. Tesfaye A.,
    6. DePinho R. A.,
    7. Alt F. W.
    (1989). IgH enhancer-mediated deregulation of N-myc gene expression in transgenic mice: generation of lymphoid neoplasias that lack c-myc expression. EMBO J 8, 1121–1128
    OpenUrlPubMedWeb of Science
    1. Frohman M. A.,
    2. Boyle M.,
    3. Martin G. R.
    (1990). Isolation of the mouse Hox-2.9 gene; Analysis of embryonic expression suggests that positional information along the anterior-posterior axis is specified by mesoderm. Development 110, 589–607
    OpenUrlAbstract/FREE Full Text
    1. Garrell J.,
    2. Campuzano S.
    (1991). The helix-loop-helix domain: A common motif for bristles, muscles and sex. BioEssays 13, 493–498
    OpenUrlCrossRefPubMed
    1. Hirning U.,
    2. Schmid P.,
    3. Schulz W. A.,
    4. Rettenberger G.,
    5. Hameister H.
    (1991). A comparative analysis of N-myc and c-myc expression and cellular proliferation in mouse organogenesis. Mech. Dev 33, 119–126
    OpenUrlCrossRefPubMedWeb of Science
    1. Ikegaki N.,
    2. Bukovsky J.,
    3. Kennett R. H.
    (1986). Identification and characterization of the NMYC gene product in human neuroblastoma cells by monoclonal antibodies with defined specificities. Proc. Natl. Acad. Sci. USA 83, 5929–5933
    OpenUrlAbstract/FREE Full Text
    1. Ikegaki N.,
    2. Kennett R. H.
    (1989). Glutaraldehyde fixation of the primary antibody-antigen complex on nitrocellulose paper increases the overall sensitivity of immunoblot assay. J. Immunol. Met 124, 205–210
    OpenUrlCrossRefPubMed
    1. Jackson T.,
    2. Allard M. F.,
    3. Sreenan C. M.,
    4. Doss L. K.,
    5. Bishop S. P.,
    6. Swain J. L.
    (1990). The c-myc proto-oncogene regulates cardiac development in transgenic mice. Mol. Cell Biol 10, 3709–3716
    OpenUrlAbstract/FREE Full Text
    1. Kato G. J.,
    2. Barrett J.,
    3. Villa-Garcia M.,
    4. Dang C. V.
    (1990). AnC. B. Moens and others499N-mycmutant miceamino-terminal c-Myc domain required for neoplastic transformation activates transcription. Mol. Cell Biol 10, 5914–5920
    OpenUrlAbstract/FREE Full Text
    1. Kato G. J.,
    2. Lee W. M. F.,
    3. Chen L.,
    4. Dang C. V.
    (1992). Max: Functional domains and interaction with c-Myc. Genes Dev 6, 81–92
    OpenUrlAbstract/FREE Full Text
    1. Katoh K.,
    2. Kanamori A.,
    3. Wakamatsu Y.,
    4. Sawai S.,
    5. Kondoh H.
    (1991). Tissue distribution of N-myc expression in the early organogenesis period of the mouse embryo. Dev. Growth Diff 33, 29–36
    OpenUrlCrossRef
    1. Kohl N. E.,
    2. Kanda N.,
    3. Schrenck R. R.,
    4. Bruns G.,
    5. Latt S. A.,
    6. Gilbert F.,
    7. Alt F. W.
    (1983). Transposition and amplification of oncogene-related sequences in human neuroblastomas. Cell 35, 359–367
    OpenUrlCrossRefPubMedWeb of Science
    1. Kretzner L.,
    2. Blackwood E. M.,
    3. Eisenman R. N.
    (1992). Myc and Max proteins possess distinct transcriptional activities. Nature 359, 426–429
    OpenUrlCrossRefPubMed
    1. Lee W. H.,
    2. Murphree A. L.,
    3. Benedict W. F.
    (1984). Expression and amplification of the N-myc gene in primary retinoblastoma. Nature 309, 458–460
    OpenUrlCrossRefPubMed
    1. Makela T. P.,
    2. Koskinen P. J.,
    3. Vastrik I.,
    4. Alitalo K.
    (1992). Alternative forms of Max as enhancers or suppressors of Myc-Ras cotransformation. Science 256, 373–377
    OpenUrlAbstract/FREE Full Text
    1. Markwald R. R.,
    2. Mjaatvedt C. H.,
    3. Krug E. L.,
    4. Sinning A. R.
    (1990). Inductive interactions in heart development: role of cardiac adherons in cushion tissue formation. Ann. NY Acad. Sci 588, 13–25
    OpenUrlPubMedWeb of Science
    1. Millauer B.,
    2. Wizigmann-Voos S.,
    3. Schnurch H.,
    4. Martinez R.,
    5. Moller N. P. H.,
    6. Risau W.,
    7. Ullrich A.
    (1993). High affinity VEGF binding and developmental expression suggest Flk-1 as a major regulator of vasculogenesis and angiogenesis. Cell 72, 835–846
    OpenUrlCrossRefPubMedWeb of Science
    1. Moens C.,
    2. Bernelot,
    3. Auerbach A. B.,
    4. Conlon R. A.,
    5. Joyner A. L.,
    6. Rossant J.
    (1992). A targeted mutation reveals a role for N-myc in branching morphogenesis in the embryonic mouse lung. Genes Dev 6, 691–704
    OpenUrlAbstract/FREE Full Text
    1. Mugrauer G.,
    2. Alt F. W.,
    3. Ekblom P.
    (1988). N-myc proto-oncogene expression during organogenesis in the developing mouse as revealed by in situ hybridization. J. Cell Biol 107, 1325–1335
    OpenUrlAbstract/FREE Full Text
    1. Mugrauer G.,
    2. Ekblom P.
    (1991). Contrasting expression patterns of three members of the myc family of protooncogenes in the developing and adult mouse kidney. J. Cell Biol 112, 13–25
    OpenUrlAbstract/FREE Full Text
    1. Mukherjee B.,
    2. Morgenbesser S. D.,
    3. DePinho R. A.
    (1992). Myc family oncoproteins function through a common pathway to transform normal cells in culture: cross-interference by Max and trans-acting dominant mutants. Genes Dev 6, 1480–1492
    OpenUrlAbstract/FREE Full Text
    1. Nau M. M.,
    2. Brooks B. J., Jr.,
    3. Carney D. N.,
    4. Gazdar A. F.,
    5. Battey J. F.,
    6. Sausville E. A.,
    7. Minna J. D.
    (1986). Human small-cell lung cancers show amplification and expression of the N-myc gene. Proc. Natl. Acad. Sci. USA 83, 1092–1096
    OpenUrlAbstract/FREE Full Text
    1. Nisen P. D.,
    2. Zimmerman K.,
    3. Cotter S. V.,
    4. Gilbert F.,
    5. Alt F. W.
    (1986). Enhanced expression of the N-myc gene in Wilms' tumours. Cancer Res 46, 6217–6222
    OpenUrlAbstract/FREE Full Text
    1. Pfeifer-Ohlsson S.,
    2. Rydnert J.,
    3. Goustin A. S.,
    4. Larsson E.,
    5. Betsholtz C.,
    6. Ohlsson R.
    (1985). Cell-type-specific pattern of myc protooncogene expression in developing human embryos. Proc. Natl. Acad. Sci. USA 82, 5050–5054
    OpenUrlAbstract/FREE Full Text
    1. Prendergast G. C.,
    2. Lawe D.,
    3. Ziff E. B.
    (1991). Association of Myn, the murine homolog of Max, with c-Myc stimulates methylation-sensitive DNA binding and Ras cotransformation. Cell 65, 395–407
    OpenUrlCrossRefPubMedWeb of Science
    1. Prendergast G. C.,
    2. Hopewell R.,
    3. Gorham B. J.,
    4. Ziff E. B.
    (1992). Biphasic effect of Max on Myc cotransformation activity and dependence on amino-and carboxy-terminal Max functions. Genes Dev 6, 2429–2439
    OpenUrlAbstract/FREE Full Text
    1. Prendergast G. C.,
    2. Ziff E. B.
    (1991). Methylation-sensitive sequence-specific DNA binding by the c-myc basic region. Science 251, 186–189
    OpenUrlAbstract/FREE Full Text
    1. Resar L. M. S.,
    2. Dolde C.,
    3. Barrett J. F.,
    4. Dang C. V.
    (1993). B-myc inhibits neoplastic transformation and transcriptional activation by c-myc. Mol. Cell Biol 13, 1130–1136
    OpenUrlAbstract/FREE Full Text
    1. Rosenbaum H.,
    2. Webb E.,
    3. Adams J. M.,
    4. Cory S.,
    5. Harris A. W.
    (1989). N-myc transgene promotes B lymphoid proliferation, elicits lymphomas and reveals cross-regulation with c-myc. EMBO J 8, 749–755
    OpenUrlPubMedWeb of Science
    1. Rumyantsev P. P.
    (1977). Interrelations of the proliferation and differentiation processes during cardiac myogenesis and regeneration. Int. Rev. Cytol 51, 187–273
    OpenUrlCrossRef
    1. Sassoon D. A.,
    2. Garner I.,
    3. Buckingham M.
    (1988). Transcripts of-cardiac and -skeletal actins are early markers for myogenesis in the mouse embryo. Development 104, 155–164
    OpenUrlAbstract
    1. Sawai S.,
    2. Shimono A.,
    3. Hanaoka K.,
    4. Kondoh H.
    (1991). Embryonic lethality resulting from disruption of both N-myc alleles in mouse zygotes. New Biologist 3, 861–869
    OpenUrlPubMedWeb of Science
    1. Sawai S.,
    2. Shimono A.,
    3. Wakamatsu Y.,
    4. Palmes C.,
    5. Hanaoka K.,
    6. Kondoh H.
    (1993). Defects of embryonic organogenesis resulting from targeted disruption of the N- myc gene in the mouse. Development 117, 1445–1455
    OpenUrlAbstract
    1. Schwab M.,
    2. Alitalo K.,
    3. Klempnauer K.,
    4. Varmus H. E.,
    5. Bishop J. M.,
    6. Gilbert F.,
    7. Brodeur G. M.,
    8. Boldstein M.,
    9. Trent J.
    (1983). Amplified DNA with limited homology to myc cellular oncogene is shared by human neuroblastoma cell lines and a neuroblastoma tumour. Nature 305, 245–248
    OpenUrlCrossRefPubMed
    1. Spooner B. S.,
    2. Wessells N. K.
    (1970). Mammalian lung development: interactions in primordium formation and bronchial morphogenesis. J. Exp. Zool 175, 445–454
    OpenUrlCrossRefPubMedWeb of Science
    1. Stanton B. R.,
    2. Reid S. W.,
    3. Parada L. F.
    (1990). Germ line transmission of an inactive N-myc allele generated by homologous recombination in mouse embryonic stem cells. Mol. Cell Biol 10, 6755–6758
    OpenUrlAbstract/FREE Full Text
    1. Stanton B. R.,
    2. Perkins A. S.,
    3. Tessarollo L.,
    4. Sassoon D. A.,
    5. Parada L. F.
    (1992). Loss of N-myc function results in embryonic lethality and failure of the epithelial component of the embryo to develop. Genes Dev 6, 2235–2247
    OpenUrlAbstract/FREE Full Text
    1. Viragh S.,
    2. Challice C. E.
    (1981). The origin of the epicardium and the embryonic myocardial circulation in the mouse. Anat. Rec 201, 157–168
    OpenUrlCrossRefPubMed
    1. Wenzel A.,
    2. Cziepluch C.,
    3. Hamann U.,
    4. Schuermann J.,
    5. Schwab M.
    (1991). The N-myc oncoprotein is associated in vivo with the phosphoprotein Max(p20/22) in human neuroblastoma cells. EMBO J 10, 3703–3712
    OpenUrlPubMedWeb of Science
    1. Wessells N. K.
    (1970). Mammalian lung development: interactions in the formation and morphogenesis of tracheal buds. J. Exp. Zool 175, 455–466
    OpenUrlCrossRefPubMedWeb of Science
    1. Wong A. J.,
    2. Ruppert J. M.,
    3. Eggleston J.,
    4. Hamilton S. R.,
    5. Baylin S. B.,
    6. Vogelstein B.
    (1986). Gene amplification of c-myc and N-myc in small cell carcinoma of the lung. Science 233, 461–464
    OpenUrlAbstract/FREE Full Text
    1. Zervos A. S.,
    2. Gyuris J.,
    3. Brent R.
    (1993). Mxi1, a protein that specifically interacts with Max to bind Myc-Max recognition sites. Cell 72, 223–232
    OpenUrlCrossRefPubMedWeb of Science
    1. Zimmerman K.,
    2. Yancopoulos G. D.,
    3. Collum R. G.,
    4. Smith R. K.,
    5. Kohl N. E.,
    6. Denis K. A.,
    7. Nau M. M.,
    8. Witte O. N.,
    9. Toran-Allerand D.,
    10. Gee C. E.,
    11. Minna J. D.,
    12. Alt F. W.
    (1986). Differential expression of myc family genes during murine development. Nature 319, 780–783
    OpenUrlCrossRefPubMed
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JOURNAL ARTICLES
Defects in heart and lung development in compound heterozygotes for two different targeted mutations at the N-myc locus
C.B. Moens, B.R. Stanton, L.F. Parada, J. Rossant
Development 1993 119: 485-499;
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