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JOURNAL ARTICLES
Pathogenesis of cleft palate in TGF-beta3 knockout mice
Y. Taya, S. O'Kane, M.W. Ferguson
Development 1999 126: 3869-3879;

Summary

We previously reported that mutation of the transforming growth factor-beta3 (TGF-beta3) gene caused cleft palate in homozygous null (−/−) mice. TGF-beta3 is normally expressed in the medial edge epithelial (MEE) cells of the palatal shelf. In the present study, we investigated the mechanisms by which TGF-beta3 deletions caused cleft palate in 129 × CF-1 mice. For organ culture, palatal shelves were dissected from embryonic day 13.5 (E13.5) mouse embryos. Palatal shelves were placed singly or in pairs on Millipore filters and cultured in DMEM/F12 medium. Shelves were placed in homologous (+/+ vs +/+, −/− vs −/−, +/− vs +/−) or heterologous (+/+ vs −/−, +/− vs −/−, +/+ vs +/−) paired combinations and examined by macroscopy and histology. Pairs of −/− and −/− shelves failed to fuse over 72 hours of culture whereas pairs of +/+ (wild-type) and +/+ or +/− (heterozygote) and +/−, as well as +/+ and −/− shelves, fused within the first 48 hour period. Histological examination of the fused +/+ and +/+ shelves showed complete disappearance of the midline epithelial seam whereas −/− and +/+ shelves still had some seam remnants. In order to investigate the ability of TGF-beta family members to rescue the fusion between −/− and −/− palatal shelves in vitro, either recombinant human (rh) TGF-beta1, porcine (p) TGF-beta2, rh TGF-beta3, rh activin, or p inhibin was added to the medium in different concentrations at specific times and for various periods during the culture. In untreated organ culture −/− palate pairs completely failed to fuse, treatment with TGF-beta3 induced complete palatal fusion, TGF-beta1 or TGF-beta2 near normal fusion, but activin and inhibin had no effect. We investigated ultrastructural features of the surface of the MEE cells using SEM to compare TGF-beta3-null embryos (E 12. 5-E 16.5) with +/+ and +/− embryos in vivo and in vitro. Up to E13.5 and after E15.5, structures resembling short rods were observed in both +/+ and −/− embryos. Just before fusion, at E14.5, a lot of filopodia-like structures appeared on the surface of the MEE cells in +/+ embryos, however, none were observed in −/− embryos, either in vivo or in vitro. With TEM these filopodia are coated with material resembling proteoglycan. Interestingly, addition of TGF-beta3 to the culture medium which caused fusion between the −/− palatal shelves also induced the appearance of these filopodia on their MEE surfaces. TGF-beta1 and TGF-beta2 also induced filopodia on the −/− MEE but to a lesser extent than TGF-beta3 and additionally induced lamellipodia on their cell surfaces. These results suggest that TGF-beta3 may regulate palatal fusion by inducing filopodia on the outer cell membrane of the palatal medial edge epithelia prior to shelf contact. Exogenous recombinant TGF-beta3 can rescue fusion in −/− palatal shelves by inducing such filopodia, illustrating that the effects of TGF-beta3 are transduced by cell surface receptors which raises interesting potential therapeutic strategies to prevent and treat embryonic cleft palate.

Reference

    1. Brunet C. L.,
    2. Sharpe P. M.,
    3. Ferguson M. W. J.
    (1995) Inhibition of TGF-3 (but not TGF- 1 or TGF-2) activity prevents normal mouse embryonic palate fusion. Int. J. Dev. Biol 39, 345–355
    OpenUrlPubMedWeb of Science
    1. Carette M. J. M.,
    2. Ferguson M. W. J.
    (1992) The fate of medial edge epithelial cells during palatal fusion in vitro: an analysis by DiI labelling and confocal microscopy. Development 114, 379–388
    OpenUrlAbstract
    1. Curtis A. S.,
    2. Buultjens T. E.
    (1973) Cell adhesion and locomotion. CIBA Found. Symp 14, 171–186
    OpenUrlPubMed
    1. Ellis I. R.,
    2. Schor S. L.
    (1998) Differential motogenic and biosynthetic response of fetal and adult skin fibroblasts to TGF-isoforms. Cytokine 10, 281–289
    OpenUrlCrossRefPubMed
    1. Ferguson M. W. J.,
    2. Honig L. S.,
    3. Slavkin H. C.
    (1984) Differentiation of cultured palatal shelves from alligator, chick, and mouse embryos. Anat. Rec 209, 231–249
    OpenUrlCrossRefPubMed
    1. Ferguson M. W. J.
    (1988) Palate development. Development 103, 41–60
    1. Ferguson M. W. J.
    (1994) Craniofacial malformations: towards a molecular understanding. Nature Genet 6, 348–356
    OpenUrlCrossRefPubMedWeb of Science
    1. Fitchett J. E.,
    2. Hay E. D.
    (1989) Medial edge epithelium transforms to mesenchyme after embryonic palatal shelves fuse. Dev. Biol 131, 455–474
    OpenUrlCrossRefPubMedWeb of Science
    1. Fitzpatrick D. R.,
    2. Denhez F.,
    3. Kondaiah P.,
    4. Akhurst R. J.
    (1990) Differential expression of TGF-isoforms in murine palatogenesis. Development 109, 585–595
    OpenUrlAbstract
    1. Gedron-Maguire M.,
    2. Mallo M.,
    3. Zhang M.,
    4. Gridley T.
    (1993) Hoxa-2 mutant mice exhibit homeotic transformation of skeletal elements derived from cranial neural crest. Cell 75, 1317–1331
    OpenUrlCrossRefPubMedWeb of Science
    1. Greene M. R.,
    2. Pratt R. M.
    (1977) Inhibition by diazo-oxo-norleucine (DON) of rat palatal glycoprotein synthesis and epithelial cell adhesion in vitro. Exp. Cell Res 105, 27–37
    OpenUrlCrossRefPubMed
    1. Griffith C. M.,
    2. Hay E. D.
    (1992) Epithelial-mesenchymal transformation during palatal fusion: carboxyfluorescein traces cells at light and electron microscopic levels. Development 116, 1087–1099
    OpenUrlAbstract/FREE Full Text
    1. Gumbiner B. M.
    (1996) Cell adhesion: The molecular basis of tissue architecture and morphogenesis. Cell 84, 345–357
    OpenUrlCrossRefPubMedWeb of Science
    1. Imamura T.,
    2. Takase M.,
    3. Nishihara A.,
    4. Oede E.,
    5. Hanai. ,
    6. Kawabata M.,
    7. Miyazono K.
    (1998) Smad6 inhibits signalling by the TGF-superfamily. Nature 389, 622–626
    1. Iozzo R. V.,
    2. Pillarisetti J.,
    3. Sharma B.,
    4. Murdoch A. D.,
    5. Danielson K. G.,
    6. Uitto J.,
    7. Mauviel A.
    (1997) Structural and functional characterization of the human perlecan gene promoter. J. Biol. Chem 272, 5219–5228
    OpenUrlAbstract/FREE Full Text
    1. Kaartinen V.,
    2. Volcken J. W.,
    3. Shuler C.,
    4. Warburton D.,
    5. Bu D.,
    6. Heisterkamp N.,
    7. Groffen J.
    (1995) Abnormal lung development andcleft palate in mice lacking TGF-3 indicates defects of epithelial-mesenchymal interaction. Nature Genet 11, 415–421
    OpenUrlCrossRefPubMedWeb of Science
    1. Kingsley D. M.
    (1994) The TGF-superfamily: new members, new receptors and new genetic tests of function in different organisms. Genes. Dev 8, 133–146
    OpenUrlFREE Full Text
    1. Kulkarni A. B.,
    2. Huh C.,
    3. Becker D.,
    4. Geiser A.,
    5. Lyght M.,
    6. Flanders K. C.,
    7. Roberts A. B.,
    8. Sporn M. B.,
    9. Ward J. M.,
    10. Karlsson S.
    (1993) Transforming growth factor-1 null mutation in mice causes excessive inflammatory response and early death. Proc. Natl. Acad. Sci. USA 90, 770–774
    OpenUrlAbstract/FREE Full Text
    1. Kurihara Y.,
    2. Kurihara H.,
    3. Suzuki H.,
    4. Kodama T.,
    5. Maemura K.,
    6. Nagai R.,
    7. Oda H.,
    8. Kuwaki T.,
    9. Cao W. H.,
    10. Kamada N.,
    11. Jishage K.,
    12. Ouchi Y.,
    13. Azuma S.,
    14. Toyoda Y.,
    15. Ishikawa T.,
    16. Kumada M.,
    17. Yazaki Y.
    (1994) Elevated blood pressure and craniofacial abnormalities in mice deficient in endothelin-1. Nature 368, 703–710
    OpenUrlCrossRefPubMed
    1. Laird P. W.,
    2. Zijderveld A.,
    3. Linders K.,
    4. Rudnicki M. A.,
    5. Jaenisch R.,
    6. Berns A.
    (1991) Simplified mammalian DNA isolation procedure. Nucl. Acids Res 19, 4293–.
    OpenUrlFREE Full Text
    1. Letterio J. J.,
    2. Geiser A. G.,
    3. Kulkarni A. B.,
    4. Roche N. S.,
    5. Sporn M. B.,
    6. Roberts A. B.
    (1994) Maternal rescue of transforming growth factor-1 null mice. Science 264, 1936–1938
    OpenUrlAbstract/FREE Full Text
    1. Lohnes D.,
    2. Kastner P.,
    3. Dierich A.,
    4. Mark M.,
    5. LeMeur M.,
    6. Chambon P.
    (1993) Function of retinoic acid receptors gamma in the mouse. Cell 73, 643–658
    OpenUrlCrossRefPubMedWeb of Science
    1. Lohnes D.,
    2. Mark M.,
    3. Mendelsohn C.,
    4. Dolle P.,
    5. Dierich A.,
    6. Gorry P.,
    7. Gansmuller A.,
    8. Chambon P.
    (1994) Function of the retinoic acid receptors (RARs) during development, (I) Craniofacial and skeletal abnormalities in RAR double mutants. Development 120, 2723–2748
    OpenUrlAbstract
    1. Martin J. F.,
    2. Bradley A.,
    3. Olsen E. N.
    (1995) The paired-like homeo box gene MHox is required for early events of skeletogenesis in multiple lineages. Genes Dev 9, 1237–1249
    OpenUrlAbstract/FREE Full Text
    1. Matzuk M. M.,
    2. Kumar T. R.,
    3. Vassalli A.,
    4. Bickenbach J. R.,
    5. Roop D. R.,
    6. Jaenisch R.,
    7. Bradley A.
    (1995) Functional analysis of activins during mammalian development. Nature 374, 354–356
    OpenUrlCrossRefPubMed
    1. Matzuk M. M.,
    2. Kumar T. R.,
    3. Bradley A.
    (1995) Different phenotypes for mice deficient in either activins or activin receptor type II. Nature 374, 356–360
    OpenUrlCrossRefPubMedWeb of Science
    1. Matzuk M. M.,
    2. Lu N.,
    3. Vogel H.,
    4. Sellheyer K.,
    5. Roop D. R.,
    6. Bradley A.
    (1995) Multiple defects and perinatal death in mice deficient in follistatin. Nature 374, 360–363
    OpenUrlCrossRefPubMed
    1. Miettinen P. J.,
    2. Chin J. R.,
    3. Shum L.,
    4. Slavkin H. C.,
    5. Shuler C. F.,
    6. Derynck R.,
    7. Werb Z.
    (1999) Epidermal growth factor receptor function is necessary for normal craniofacial development and palate closure. Nature Genet 22, 69–73
    OpenUrlCrossRefPubMedWeb of Science
    1. Mo R.,
    2. Freer A. M.,
    3. Zinyk D. L.,
    4. Crackower M. A.,
    5. Michaud J.,
    6. Heng H. H.,
    7. Chik K. W.,
    8. Shi X. M.,
    9. Tsui L. C.,
    10. Cheng S. H.,
    11. Joyner A. L.,
    12. Hui C.
    (1997) Specific and redundant function of Gli2 and Gli3 zinc finger genes in skeletal patterning and development. Development 124, 113–123
    OpenUrlAbstract
    1. Nakao A.,
    2. Afrakhte M.,
    3. Moren A.,
    4. Nakajima T.,
    5. Christian J. L.,
    6. Heuchel R.,
    7. Itoh S.,
    8. Kawabata M.,
    9. Heldin N. E.,
    10. Heldin C. H.,
    11. ten Dijke P.
    (1997) Identification of smad7, a TGF-inducible antagonist of TGF- signalling. Nature 389, 631–635
    OpenUrlCrossRefPubMedWeb of Science
    1. Nobes C. D.,
    2. Hall A.
    (1995) Rho, rac, and cdc42 GTPases regulate the assembly of multimolecular focal complexs associated with actin stress fibers, lamellipodia, and filopodia. Cell 81, 53–62
    OpenUrlCrossRefPubMedWeb of Science
    1. Orioli D.,
    2. Henkemeyer M.,
    3. Lemke G.,
    4. Klein R.,
    5. Pawson T.
    (1996) Sek4 and nuk receptors cooperate in guidance of commissural axons and in palate formation. EMBO J 15, 6035–6049
    OpenUrlPubMedWeb of Science
    1. Pelton R. W.,
    2. Hogan B. L. M.,
    3. Miller D. A.,
    4. Moses H. L.
    (1990) Differential expression of genes encoding TGF-1, 2 and3 during murine palate formation. Dev. Biol 141, 456–460
    OpenUrlCrossRefPubMedWeb of Science
    1. Peters H.,
    2. Neubuser A.,
    3. Balling R.
    (1997) The role of Pax9 during mouse development. Dev. Biol 186, 333–.
    1. Proetzel G.,
    2. Pawlowski S. A.,
    3. Wiles M. V.,
    4. Yin M.,
    5. Boivin G. P.,
    6. Howles P. N.,
    7. Ding J.,
    8. Ferguson M. W. J.,
    9. Doetschman T.
    (1995) Transformating growth factor-3 is required for secondary palate fusion. Nature Genet 11, 409–414
    OpenUrlCrossRefPubMedWeb of Science
    1. Ramsdell A. F.,
    2. Markwald R. R.
    (1997) Induction of endocardial cushion tissue in the avian heart is regulated, in part, by TGF beta-3-mediated autocrine signalling. Dev. Biol 188, 64–74
    OpenUrlCrossRefPubMedWeb of Science
    1. Rijli F. M.,
    2. Mark M.,
    3. Lakkaraju S.,
    4. Dierich A.,
    5. Dolle P.,
    6. Chambon P.
    (1993) A homeotic transformation is generated in the rostral branchial region of the head by disruption of Hoxa-2, which acts as a selector gene. Cell 75, 1333–1349
    OpenUrlCrossRefPubMedWeb of Science
    1. Sanford L. P.,
    2. Ormsby I.,
    3. Groot A. C. G.,
    4. Sarinola H.,
    5. Friedman R.,
    6. Boivin G. P.,
    7. Cardell E. L.,
    8. Doetschman T.
    (1997) TGF-2 knockout mice have multiple developmental defects that are non-overlapping with other TGF- knockout phenotype. Development 124, 2659–2670
    OpenUrlAbstract
    1. Satokata I.,
    2. Maas R.
    (1994) Msx1 deficient mice exhibit cleft palate and abnormalities of craniofacial and tooth development. Nature Genet 6, 348–355
    1. Schupback P. M.,
    2. Schroeder H. E.
    (1986) Fate of unfused medial edge epithelia in rat fetuses with experimentally induced cleft palate: 1. from 16.3 to 17.7 days of gestation. J. Craniofacial. Genet 2, 293–318
    OpenUrl
    1. Shah M.,
    2. Foreman D. M.,
    3. Ferguson M. W. J.
    (1995) Neutralisation of TGF-1 and TGF- 2 or exogenous addition of TGF-3 to cutaneous rat wounds reduces scarring. J. Cell Sci 108, 985–1002
    OpenUrlAbstract/FREE Full Text
    1. Shuler C. F.,
    2. Guo Y.,
    3. Majumder A.,
    4. Luo R.
    (1991) Molecular and morphologic changes during the epithelial-mesenchymal transformation of palatal shelf medial edge epithelium in vitro. Int. J. Dev. Biol 35, 463–472
    OpenUrlPubMed
    1. Shuler C. F.,
    2. Halpern D. E.,
    3. Guo Y.,
    4. Sank A. C.
    (1992) Medial edge epithelium fate traced by cell lineage analysis during epithelial-mesenchymal transformation in vivo. Dev. Biol 154, 318–330
    OpenUrlCrossRefPubMedWeb of Science
    1. Shull M. M.,
    2. Ormsby I.,
    3. Kier A. B.,
    4. Pawlowski S.,
    5. Diebold R. J.,
    6. Yin M.,
    7. Allen R.,
    8. Sidman C.,
    9. Proetzel G.,
    10. Calvin D.,
    11. Annunziata N.,
    12. Doetschman T.
    (1992) Targeted disruption of the mouse transforming growth factor-1 gene results in multifocal inflammatory disease. Nature 359, 693–699
    OpenUrlCrossRefPubMedWeb of Science
    1. Sun D.,
    2. Vandergurg C. R.,
    3. Odierna G. S.,
    4. Hay E.
    (1998) TGF-3 promotes transformation of chicken palate medial edge epithelium to mesenchyme in vitro. Development 125, 95–105
    OpenUrlAbstract
    1. Takagi T.,
    2. Moribe H.,
    3. Kondoh H.,
    4. Higashi Y.
    (1998) deltaEF1, a zinc finger and homeodomein transcription factor, is required for skelton patterning in multi lineages. Development 125, 21–31
    OpenUrlAbstract
    1. Takihara Y.,
    2. Tomotsune D.,
    3. Shirai M.,
    4. Katoh F. Y.,
    5. Nishii K.,
    6. Motaleb M. A.,
    7. Nomura M.,
    8. Tsuchiya R.,
    9. Fujita Y.,
    10. Shibata Y.,
    11. Higashinakagawa T.,
    12. Shimada K.
    (1997) Targeted distruption of the mouse homolog of the Drosophila polyhomeotic gene leads to altered anteroposterior patterning and neural crest defects. Development 124, 3673–3682
    OpenUrlAbstract
    1. Tissier-Seta J.-P.,
    2. Mucchielli M.-L.,
    3. Mark M.,
    4. Mattei M.-G.,
    5. Goridis C.,
    6. Brunet J.-F.
    (1995) Barx1, a new mouse homeodomain transcription factor expressed in cranio-facial ectomesenchyme and the stomach. Mech. Dev 51, 3–15
    OpenUrlCrossRefPubMedWeb of Science
    1. Waterman R. E.,
    2. Ross L. M.,
    3. Meller S. M.
    (1973) Alternations in the epithelial surface of A/Jax mouse palatal shelves prior to and during of palatal fusion: a scanning electron microscopic study. Anat. Rec 176, 361–376
    OpenUrlCrossRefPubMed
    1. Winograd J.,
    2. Reilly M. P.,
    3. Roe R.,
    4. Lutz J.,
    5. Laughner E.,
    6. Xu X.,
    7. Hu L.,
    8. Asakura T.,
    9. vander Kolk C.,
    10. Strandberg J. D.,
    11. Semenza G. L.
    (1997) Perinatal lethality and multiple craniofacial malformations in MSX2 transgenic mice. Human Mol. Genet 6, 369–379
    OpenUrlAbstract/FREE Full Text
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JOURNAL ARTICLES
Pathogenesis of cleft palate in TGF-beta3 knockout mice
Y. Taya, S. O'Kane, M.W. Ferguson
Development 1999 126: 3869-3879;
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