46 results on '"Ciruna B"'
Search Results
2. Loss-of-function mutations in UDP-Glucose 6-Dehydrogenase cause recessive developmental epileptic encephalopathy
- Author
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Hengel, H. (Holger), Bosso-Lefèvre, C. (Célia), Grady, G. (George), Szenker-Ravi, E. (Emmanuelle), Li, H. (Hankun), Pierce, S. (Sarah), Lebigot, É. (Élise), Tan, T.-T. (Thong-Teck), Eio, M.Y. (Michelle Y.), Narayanan, G. (Gunaseelan), Utami, K.H. (Kagistia Hana), Yau, M. (Monica), Handal, N. (Nader), Deigendesch, W. (Werner), Keimer, R. (Reinhard), Marzouqa, H.M. (Hiyam M.), Gunay-Aygun, M. (Meral), Muriello, M.J. (Michael J.), Verhelst, H. (H.), Weckhuysen, S. (Sarah), Mahida, S. (Sonal), Naidu, S. (Sakkubai), Thomas, T.G. (Terrence G.), Lim, J.Y. (Jiin Ying), Tan, E.S. (Ee Shien), Haye, D. (Damien), Willemsen, M.A. (Michél), Oegema, R. (Renske), Mitchell, W.G. (Wendy G.), Pierson, T.M. (Tyler Mark), Andrews, M.V. (Marisa V.), Willing, M.C. (Marcia C.), Rodan, L.H. (Lance H.), Barakat, T.S. (Tahsin Stefan), Slegtenhorst, M.A. (Marjon) van, Gavrilova, R.H. (Ralitza H.), Martinelli, D. (Diego), Gilboa, T. (Tal), Tamim, A.M. (Abdullah M.), Hashem, M.O. (Mais O.), AlSayed, M.D. (Moeenaldeen D.), Abdulrahim, M.M. (Maha M.), Al-Owain, M. (Mohammed), Awaji, A. (Ali), Mahmoud, A.A.H. (Adel A. H.), Faqeih, E.A. (Eissa A.), Asmari, A.A. (Ali Al), Algain, S.M. (Sulwan M.), Jad, L.A. (Lamyaa A.), Aldhalaan, H.M. (Hesham M.), Helbig, I. (Ingo), Koolen, D.A. (David), Rieß, A. (Angelika), Kraegeloh-Mann, I. (Ingeborg), Bauer, P. (Peter), Gulsuner, S. (Suleyman), Stamberger, H. (Hannah), Ng, A.Y.J. (Alvin Yu Jin), Tang, S. (Sha), Tohari, S. (Sumanty), Keren, B. (Boris), Schultz-Rogers, L.E. (Laura E.), Klee, E.W. (Eric W.), Barresi, S. (Sabina), Tartaglia, M. (Marco), Mor-Shaked, H. (Hagar), Maddirevula, S. (Sateesh), Begtrup, A. (Amber), Telegrafi, A. (Aida), Pfundt, R. (Rolph), Schüle, R. (Rebecca), Ciruna, B. (Brian), Bonnard, C. (Carine), Pouladi, M.A. (Mahmoud A.), Stewart, J.C. (James C.), Claridge-Chang, A. (Adam), Lefeber, D.J. (Dirk J.), Alkuraya, F.S. (Fowzan S), Mathuru, A.S. (Ajay S.), Venkatesh, B. (Byrappa), Barycki, J.J. (Joseph J.), Simpson, M.A. (Melanie A.), Jamuar, S.S. (Saumya S.), Schöls, L. (Ludger), Reversade, B. (Bruno), Hengel, H. (Holger), Bosso-Lefèvre, C. (Célia), Grady, G. (George), Szenker-Ravi, E. (Emmanuelle), Li, H. (Hankun), Pierce, S. (Sarah), Lebigot, É. (Élise), Tan, T.-T. (Thong-Teck), Eio, M.Y. (Michelle Y.), Narayanan, G. (Gunaseelan), Utami, K.H. (Kagistia Hana), Yau, M. (Monica), Handal, N. (Nader), Deigendesch, W. (Werner), Keimer, R. (Reinhard), Marzouqa, H.M. (Hiyam M.), Gunay-Aygun, M. (Meral), Muriello, M.J. (Michael J.), Verhelst, H. (H.), Weckhuysen, S. (Sarah), Mahida, S. (Sonal), Naidu, S. (Sakkubai), Thomas, T.G. (Terrence G.), Lim, J.Y. (Jiin Ying), Tan, E.S. (Ee Shien), Haye, D. (Damien), Willemsen, M.A. (Michél), Oegema, R. (Renske), Mitchell, W.G. (Wendy G.), Pierson, T.M. (Tyler Mark), Andrews, M.V. (Marisa V.), Willing, M.C. (Marcia C.), Rodan, L.H. (Lance H.), Barakat, T.S. (Tahsin Stefan), Slegtenhorst, M.A. (Marjon) van, Gavrilova, R.H. (Ralitza H.), Martinelli, D. (Diego), Gilboa, T. (Tal), Tamim, A.M. (Abdullah M.), Hashem, M.O. (Mais O.), AlSayed, M.D. (Moeenaldeen D.), Abdulrahim, M.M. (Maha M.), Al-Owain, M. (Mohammed), Awaji, A. (Ali), Mahmoud, A.A.H. (Adel A. H.), Faqeih, E.A. (Eissa A.), Asmari, A.A. (Ali Al), Algain, S.M. (Sulwan M.), Jad, L.A. (Lamyaa A.), Aldhalaan, H.M. (Hesham M.), Helbig, I. (Ingo), Koolen, D.A. (David), Rieß, A. (Angelika), Kraegeloh-Mann, I. (Ingeborg), Bauer, P. (Peter), Gulsuner, S. (Suleyman), Stamberger, H. (Hannah), Ng, A.Y.J. (Alvin Yu Jin), Tang, S. (Sha), Tohari, S. (Sumanty), Keren, B. (Boris), Schultz-Rogers, L.E. (Laura E.), Klee, E.W. (Eric W.), Barresi, S. (Sabina), Tartaglia, M. (Marco), Mor-Shaked, H. (Hagar), Maddirevula, S. (Sateesh), Begtrup, A. (Amber), Telegrafi, A. (Aida), Pfundt, R. (Rolph), Schüle, R. (Rebecca), Ciruna, B. (Brian), Bonnard, C. (Carine), Pouladi, M.A. (Mahmoud A.), Stewart, J.C. (James C.), Claridge-Chang, A. (Adam), Lefeber, D.J. (Dirk J.), Alkuraya, F.S. (Fowzan S), Mathuru, A.S. (Ajay S.), Venkatesh, B. (Byrappa), Barycki, J.J. (Joseph J.), Simpson, M.A. (Melanie A.), Jamuar, S.S. (Saumya S.), Schöls, L. (Ludger), and Reversade, B. (Bruno)
- Abstract
Developmental epileptic encephalopathies are devastating disorders characterized by intractable epileptic seizures and developmental delay. Here, we report an allelic series of germline recessive mutations in UGDH in 36 cases from 25 families presenting with epileptic encephalopathy with developmental delay and hypotonia. UGDH encodes an oxidoreductase that converts UDP-glucose to UDP-glucuronic acid, a key component of specific proteoglycans and glycolipids. Consistent with being loss-of-function alleles, we show using patients’ primary fibroblasts and biochemical assays, that these mutations either impair UGDH stability, oligomerization, or enzymatic activity. In vitro, patient-derived cerebral organoids are smaller with a reduced number of proliferating neuronal progenitors while mutant ugdh zebrafish do not phenocopy the human disease. Our study defines UGDH as a key player for the production of extracellular matrix components that are essential for human brain development. Based on the incidence of variants observed, UGDH mutations are likely to be a frequent cause of recessive epileptic encephalopathy.
- Published
- 2020
- Full Text
- View/download PDF
3. Loss-of-function mutations in UDP-Glucose 6-Dehydrogenase cause recessive developmental epileptic encephalopathy
- Author
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Hengel, H., Bosso-Lefevre, C., Grady, G., Szenker-Ravi, E., Li, H., Pierce, S., Lebigot, E., Tan, T.T., Eio, M.Y., Narayanan, G., Utami, K.H., Yau, M., Handal, N., Deigendesch, W., Keimer, R., Marzouqa, H.M., Gunay-Aygun, M., Muriello, M.J., Verhelst, H., Weckhuysen, S., Mahida, S., Naidu, S., Thomas, T.G., Lim, J.Y., Tan, E.S., Haye, D., Willemsen, M.A.A.P., Oegema, R., Mitchell, W.G., Pierson, T.M., Andrews, M.V., Willing, M.C., Rodan, L.H., Barakat, T.S., Slegtenhorst, M. van, Gavrilova, R.H., Martinelli, D., Gilboa, T., Tamim, A.M., Hashem, M.O., AlSayed, M.D., Abdulrahim, M.M., Al-Owain, M., Awaji, A., Mahmoud, A.A.H., Faqeih, E.A., Asmari, A.A., Algain, S.M., Jad, L.A., Aldhalaan, H.M., Helbig, I., Koolen, D.A., Riess, A., Kraegeloh-Mann, I., Bauer, P., Gulsuner, S., Stamberger, H., Ng, A.Y., Tang, S., Tohari, S., Keren, B., Schultz-Rogers, L.E., Klee, E.W., Barresi, S., Tartaglia, M., Mor-Shaked, H., Maddirevula, S., Begtrup, A., Telegrafi, A., Pfundt, R.P., Schule, R., Ciruna, B., Bonnard, C., Pouladi, M.A., Stewart, J.C., Claridge-Chang, A., Lefeber, D.J., Alkuraya, F.S., Mathuru, A.S., Venkatesh, B., Barycki, J.J., Simpson, Malcolm, Jamuar, S.S., Schols, L., Reversade, B., Hengel, H., Bosso-Lefevre, C., Grady, G., Szenker-Ravi, E., Li, H., Pierce, S., Lebigot, E., Tan, T.T., Eio, M.Y., Narayanan, G., Utami, K.H., Yau, M., Handal, N., Deigendesch, W., Keimer, R., Marzouqa, H.M., Gunay-Aygun, M., Muriello, M.J., Verhelst, H., Weckhuysen, S., Mahida, S., Naidu, S., Thomas, T.G., Lim, J.Y., Tan, E.S., Haye, D., Willemsen, M.A.A.P., Oegema, R., Mitchell, W.G., Pierson, T.M., Andrews, M.V., Willing, M.C., Rodan, L.H., Barakat, T.S., Slegtenhorst, M. van, Gavrilova, R.H., Martinelli, D., Gilboa, T., Tamim, A.M., Hashem, M.O., AlSayed, M.D., Abdulrahim, M.M., Al-Owain, M., Awaji, A., Mahmoud, A.A.H., Faqeih, E.A., Asmari, A.A., Algain, S.M., Jad, L.A., Aldhalaan, H.M., Helbig, I., Koolen, D.A., Riess, A., Kraegeloh-Mann, I., Bauer, P., Gulsuner, S., Stamberger, H., Ng, A.Y., Tang, S., Tohari, S., Keren, B., Schultz-Rogers, L.E., Klee, E.W., Barresi, S., Tartaglia, M., Mor-Shaked, H., Maddirevula, S., Begtrup, A., Telegrafi, A., Pfundt, R.P., Schule, R., Ciruna, B., Bonnard, C., Pouladi, M.A., Stewart, J.C., Claridge-Chang, A., Lefeber, D.J., Alkuraya, F.S., Mathuru, A.S., Venkatesh, B., Barycki, J.J., Simpson, Malcolm, Jamuar, S.S., Schols, L., and Reversade, B.
- Abstract
Contains fulltext : 218288.pdf (publisher's version ) (Open Access), Developmental epileptic encephalopathies are devastating disorders characterized by intractable epileptic seizures and developmental delay. Here, we report an allelic series of germline recessive mutations in UGDH in 36 cases from 25 families presenting with epileptic encephalopathy with developmental delay and hypotonia. UGDH encodes an oxidoreductase that converts UDP-glucose to UDP-glucuronic acid, a key component of specific proteoglycans and glycolipids. Consistent with being loss-of-function alleles, we show using patients' primary fibroblasts and biochemical assays, that these mutations either impair UGDH stability, oligomerization, or enzymatic activity. In vitro, patient-derived cerebral organoids are smaller with a reduced number of proliferating neuronal progenitors while mutant ugdh zebrafish do not phenocopy the human disease. Our study defines UGDH as a key player for the production of extracellular matrix components that are essential for human brain development. Based on the incidence of variants observed, UGDH mutations are likely to be a frequent cause of recessive epileptic encephalopathy.
- Published
- 2020
4. Loss-of-function mutations in UDP-Glucose 6-Dehydrogenase cause recessive developmental epileptic encephalopathy
- Author
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Reversade, Bruno, Hengel, H.; Bosso-Lefèvre, C.; Grady, G.; Szenker-Ravi, E.; Li, H.; Pierce, S.; Lebigot, É.; Tan, T.-T.; Eio, M.Y.; Narayanan, G.; Utami, K.H.; Yau, M.; Handal, N.; Deigendesch, W.; Keimer, R.; Marzouqa, H.M.; Gunay-Aygun, M.; Muriello, M.J.; Verhelst, H.; Weckhuysen, S.; Mahida, S.; Naidu, S.; Thomas, T.G.; Lim, J.Y.; Tan, E.S.; Haye, D.; Willemsen, M.A.A.P.; Oegema, R.; Mitchell, W.G.; Pierson, T.M.; Andrews, M.V.; Willing, M.C.; Rodan, L.H.; Barakat, T.S.; van Slegtenhorst, M.; Gavrilova, R.H.; Martinelli, D.; Gilboa, T.; Tamim, A.M.; Hashem, M.O.; AlSayed, M.D.; Abdulrahim, M.M.; Al-Owain, M.; Awaji, A.; Mahmoud, A.A.H.; Faqeih, E.A.; Asmari, A.A.; Algain, S.M.; Jad, L.A.; Aldhalaan, H.M.; Helbig, I.; Koolen, D.A.; Riess, A.; Kraegeloh-Mann, I.; Bauer, P.; Gulsuner, S.; Stamberger, H.; Ng, A.Y.J.; Tang, S.; Tohari, S.; Keren, B.; Schultz-Rogers, L.E.; Klee, E.W.; Barresi, S.; Tartaglia, M.; Mor-Shaked, H.; Maddirevula, S.; Begtrup, A.; Telegrafi, A.; Pfundt, R.; Schüle, R.; Ciruna, B.; Bonnard, C.; Pouladi, M.A.; Stewart, J.C.; Claridge-Chang, A.; Lefeber, D.J.; Alkuraya, F.S.; Mathuru, A.S.; Venkatesh, B.; Barycki, J.J.; Simpson, M.A.; Jamuar, S.S.; Schöls, L, School of Medicine, Reversade, Bruno, Hengel, H.; Bosso-Lefèvre, C.; Grady, G.; Szenker-Ravi, E.; Li, H.; Pierce, S.; Lebigot, É.; Tan, T.-T.; Eio, M.Y.; Narayanan, G.; Utami, K.H.; Yau, M.; Handal, N.; Deigendesch, W.; Keimer, R.; Marzouqa, H.M.; Gunay-Aygun, M.; Muriello, M.J.; Verhelst, H.; Weckhuysen, S.; Mahida, S.; Naidu, S.; Thomas, T.G.; Lim, J.Y.; Tan, E.S.; Haye, D.; Willemsen, M.A.A.P.; Oegema, R.; Mitchell, W.G.; Pierson, T.M.; Andrews, M.V.; Willing, M.C.; Rodan, L.H.; Barakat, T.S.; van Slegtenhorst, M.; Gavrilova, R.H.; Martinelli, D.; Gilboa, T.; Tamim, A.M.; Hashem, M.O.; AlSayed, M.D.; Abdulrahim, M.M.; Al-Owain, M.; Awaji, A.; Mahmoud, A.A.H.; Faqeih, E.A.; Asmari, A.A.; Algain, S.M.; Jad, L.A.; Aldhalaan, H.M.; Helbig, I.; Koolen, D.A.; Riess, A.; Kraegeloh-Mann, I.; Bauer, P.; Gulsuner, S.; Stamberger, H.; Ng, A.Y.J.; Tang, S.; Tohari, S.; Keren, B.; Schultz-Rogers, L.E.; Klee, E.W.; Barresi, S.; Tartaglia, M.; Mor-Shaked, H.; Maddirevula, S.; Begtrup, A.; Telegrafi, A.; Pfundt, R.; Schüle, R.; Ciruna, B.; Bonnard, C.; Pouladi, M.A.; Stewart, J.C.; Claridge-Chang, A.; Lefeber, D.J.; Alkuraya, F.S.; Mathuru, A.S.; Venkatesh, B.; Barycki, J.J.; Simpson, M.A.; Jamuar, S.S.; Schöls, L, and School of Medicine
- Abstract
Developmental epileptic encephalopathies are devastating disorders characterized by intractable epileptic seizures and developmental delay. Here, we report an allelic series of germline recessive mutations in UGDH in 36 cases from 25 families presenting with epileptic encephalopathy with developmental delay and hypotonia. UGDH encodes an oxidoreductase that converts UDP-glucose to UDP-glucuronic acid, a key component of specific proteoglycans and glycolipids. Consistent with being loss-of-function alleles, we show using patients’ primary fibroblasts and biochemical assays, that these mutations either impair UGDH stability, oligomerization, or enzymatic activity. In vitro, patient-derived cerebral organoids are smaller with a reduced number of proliferating neuronal progenitors while mutant ugdh zebrafish do not phenocopy the human disease. Our study defines UGDH as a key player for the production of extracellular matrix components that are essential for human brain development. Based on the incidence of variants observed, UGDH mutations are likely to be a frequent cause of recessive epileptic encephalopathy., German Research Foundation (DFG); European Union (European Union); NEUROMICS Network; International Coordination Action (ICA); Fund for Scientific Research Flanders (FWO); Netherlands Organization for Scientific Research (ZONMW VIDI); National Medical Research Council, Singapore; A Strategic Positioning Fund on Genetic Orphan Diseases (GODAFIT); Industry Alignment Fund on Singapore Childhood Undiagnosed Diseases Program (SUREKids); Biomedical Research Council, A*STAR; Diana and Steve Marienhoff Fashion Industries Guild Endowed Fellowship in Pediatric Neuromuscular Diseases; Fondazione Bambino Gesù (Vite Coraggiose); Canadian Institutes of Health Research; Natural Sciences and Engineering Research Council of Canada; Eurocores Program EuroEPINOMICS; University of Antwerp Research Fund; FRAXA Foundation; Brain & Behavior Research Foundation, NARSAD Young Investigator Grant
- Published
- 2020
5. Neuroinflammatory signals drive spinal curve formation in zebrafish models of idiopathic scoliosis
- Author
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Van Gennip, J. L. M., primary, Boswell, C. W., additional, and Ciruna, B., additional
- Published
- 2018
- Full Text
- View/download PDF
6. Investigating the link between PCP signalling and ciliogenesis during zebrafish development
- Author
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Borovina, A, primary and Ciruna, B, additional
- Published
- 2012
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7. Automated batch transfer of zebrafish embryos using a multi-degrees-of-freedom system.
- Author
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Xuping Zhang, Zhe Lu, Gelinas, D., Ciruna, B., and Yu Sun
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- 2011
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8. ABSTRACT Conserved pathways of FGF action in trophoblast and mesoderm development
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Rossant, J, primary, Ciruna, B, additional, Kunath, T, additional, Mao, C -A, additional, and Strumpf, D, additional
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- 2000
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9. Expression of the T-box gene Eomesodemin during early mouse development
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Ciruna, B. G. and Rossant, J.
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- 1999
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10. Chimeric analysis of fibroblast growth factor receptor-1 (Fgfr1) function: a role for FGFR1 in morphogenetic movement through the primitive streak.
- Author
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Ciruna, B G, Schwartz, L, Harpal, K, Yamaguchi, T P, and Rossant, J
- Abstract
Fibroblast growth factor (FGF) signaling has been implicated in the patterning of mesoderm and neural lineages during early vertebrate development. In the mouse, FGF receptor-1 (FGFR1) is expressed in an appropriate spatial and temporal manner to be orchestrating these functions. Mouse embryos homozygous for a mutated Fgfr1 allele (fgfr1(delta tmk)) die early in development, show abnormal growth and aberrant mesodermal patterning. We have performed a chimeric analysis to further study FGFR1 function in the morphogenesis and patterning of the mesodermal germ layer at gastrulation. At E9.5, fgfr1(delta tmk)/fgfr1(delta tmk) cells showed a marked deficiency in their ability to contribute to the extra-embryonic, cephalic, heart, axial and paraxial mesoderm, and to the endoderm of chimeric embryos. Analysis at earlier stages of development revealed that fgfr1(delta tmk)/fgfr1(delta tmk) cells accumulated within the primitive streak of chimeric embryos, and consequently failed to populate the anterior mesoderm and endodermal lineages at their inception. We suggest that the primary defect associated with the fgfr1(delta tmk) mutation is a deficiency in the ability of epiblast cells to traverse the primitive streak. fgfr1(delta tmk)/fgfr1(delta tmk) cells that accumulated within the primitive streak of chimeric embryos tended to form secondary neural tubes. These secondary neural tubes were entirely fgfr1(delta tmk)/fgfr1(delta tmk) cell derived. The adoption of ectopic neural fate suggests that normal morphogenetic movement through the streak is essential not only for proper mesodermal patterning but also for correct determination of mesodermal/neurectodermal cell fates.
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- 1997
11. FGF Signaling Regulates Mesoderm Cell Fate Specification and Morphogenetic Movement at the Primitive Streak
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CIRUNA, B
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- 2001
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12. Loss-of-function mutations in UDP-Glucose 6-Dehydrogenase cause recessive developmental epileptic encephalopathy
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Ali Al Asmari, Emmanuelle Szenker-Ravi, Carine Bonnard, Bruno Reversade, Laura Schultz-Rogers, I. Kraegeloh-Mann, Maha Abdulrahim, Hesham Aldhalaan, Byrappa Venkatesh, Célia Bosso-Lefèvre, Aida Telegrafi, Hiyam M. Marzouqa, Gunaseelan Narayanan, Sha Tang, Sonal Mahida, Melanie A. Simpson, Fowzan S. Alkuraya, Michelle Eio, Eissa Faqeih, Renske Oegema, Sarah Weckhuysen, George Grady, Joseph J. Barycki, Mohammed Al-Owain, Lamyaa A. Jad, David A. Koolen, Marjon van Slegtenhorst, Tyler Mark Pierson, Marisa V. Andrews, Rebecca Schüle, Reinhard Keimer, Amber Begtrup, Sateesh Maddirevula, Michael Muriello, Sakkubai Naidu, Damien Haye, Adel A H Mahmoud, Brian Ciruna, Abdullah Tamim, Thong Teck Tan, Rolph Pfundt, Peter Bauer, Jiin Ying Lim, Ali Awaji, Marco Tartaglia, Meral Gunay-Aygun, Eric W. Klee, Marcia C. Willing, Monica Yau, Angelika Riess, Diego Martinelli, Sabina Barresi, Sumanty Tohari, Werner Deigendesch, Dirk Lefeber, Saumya Shekhar Jamuar, Ludger Schöls, Ralitza H. Gavrilova, Alvin Yu Jin Ng, Hannah Stamberger, Suleyman Gulsuner, Adam Claridge-Chang, Élise Lebigot, Moeenaldeen Al-Sayed, Ee Shien Tan, Kagistia Hana Utami, Sarah B. Pierce, Helene Verhelst, Hankun Li, James C. Stewart, Ingo Helbig, Tal Gilboa, Mahmoud A. Pouladi, Hagar Mor-Shaked, Boris Keren, Ajay S. Mathuru, Holger Hengel, Michèl A.A.P. Willemsen, Nader Handal, Tahsin Stefan Barakat, Sulwan M. Algain, Terrence Thomas, Lance H. Rodan, Mais Hashem, Wendy G. Mitchell, Center for Reproductive Medicine, ARD - Amsterdam Reproduction and Development, ACS - Diabetes & metabolism, Clinical Genetics, Reversade, Bruno, Hengel, H., Bosso-Lefèvre, C., Grady, G., Szenker-Ravi, E., Li, H., Pierce, S., Lebigot, É., Tan, T.-T., Eio, M.Y., Narayanan, G., Utami, K.H., Yau, M., Handal, N., Deigendesch, W., Keimer, R., Marzouqa, H.M., Gunay-Aygun, M., Muriello, M.J., Verhelst, H., Weckhuysen, S., Mahida, S., Naidu, S., Thomas, T.G., Lim, J.Y., Tan, E.S., Haye, D., Willemsen, M.A.A.P., Oegema, R., Mitchell, W.G., Pierson, T.M., Andrews, M.V., Willing, M.C., Rodan, L.H., Barakat, T.S., van Slegtenhorst, M., Gavrilova, R.H., Martinelli, D., Gilboa, T., Tamim, A.M., Hashem, M.O., AlSayed, M.D., Abdulrahim, M.M., Al-Owain, M., Awaji, A., Mahmoud, A.A.H., Faqeih, E.A., Asmari, A.A., Algain, S.M., Jad, L.A., Aldhalaan, H.M., Helbig, I., Koolen, D.A., Riess, A., Kraegeloh-Mann, I., Bauer, P., Gulsuner, S., Stamberger, H., Ng, A.Y.J., Tang, S., Tohari, S., Keren, B., Schultz-Rogers, L.E., Klee, E.W., Barresi, S., Tartaglia, M., Mor-Shaked, H., Maddirevula, S., Begtrup, A., Telegrafi, A., Pfundt, R., Schüle, R., Ciruna, B., Bonnard, C., Pouladi, M.A., Stewart, J.C., Claridge-Chang, A., Lefeber, D.J., Alkuraya, F.S., Mathuru, A.S., Venkatesh, B., Barycki, J.J., Simpson, M.A., Jamuar, S.S., Schöls, L, and School of Medicine
- Subjects
0301 basic medicine ,Male ,Glycobiology ,General Physics and Astronomy ,VARIANTS ,Encephalopathy ,Neurodegenerative ,Germline ,0302 clinical medicine ,UDP-GLUCOSE DEHYDROGENASE ,Loss of Function Mutation ,Medicine and Health Sciences ,EMBRYOGENESIS ,2.1 Biological and endogenous factors ,UGDH protein, human ,Aetiology ,Child ,lcsh:Science ,Zebrafish ,UTILITY ,Genetics ,pathology [Organoids] ,Multidisciplinary ,Uridine diphosphate glucose dehydrogenase ,Uridine diphosphate ,DP-glucuronic acid ,Syndrome ,Disorders of movement Donders Center for Medical Neuroscience [Radboudumc 3] ,Hypotonia ,3. Good health ,Pedigree ,DEFICIENCY ,genetics [Loss of Function Mutation] ,Organoids ,genetics [Uridine Diphosphate Glucose Dehydrogenase] ,Child, Preschool ,Neurological ,Medicine ,Female ,ddc:500 ,medicine.symptom ,Oxidoreductases ,Engineering sciences. Technology ,Rare cancers Radboud Institute for Health Sciences [Radboudumc 9] ,ENZYME ,Adolescent ,CONGENITAL DISORDER ,Science ,Intellectual and Developmental Disabilities (IDD) ,genetics [Epilepsy] ,chemistry [Oxidoreductases] ,Genetics and Molecular Biology ,Genes, Recessive ,Biology ,Uridine Diphosphate Glucose Dehydrogenase ,Article ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Protein Domains ,medicine ,Animals ,Humans ,Recessive ,Clinical genetics ,Allele ,Preschool ,Gene ,Loss function ,Alleles ,HEPARAN-SULFATE ,Phenocopy ,genetics [Oxidoreductases] ,Neurodevelopmental disorders Donders Center for Medical Neuroscience [Radboudumc 7] ,Epilepsy ,GLYCOSYLATION ,Neurosciences ,Infant ,General Chemistry ,biology.organism_classification ,medicine.disease ,Brain Disorders ,carbohydrates (lipids) ,Kinetics ,030104 developmental biology ,Genes ,General Biochemistry ,Neuronal development ,lcsh:Q ,Human medicine ,030217 neurology & neurosurgery ,Congenital disorder - Abstract
Developmental epileptic encephalopathies are devastating disorders characterized by intractable epileptic seizures and developmental delay. Here, we report an allelic series of germline recessive mutations in UGDH in 36 cases from 25 families presenting with epileptic encephalopathy with developmental delay and hypotonia. UGDH encodes an oxidoreductase that converts UDP-glucose to UDP-glucuronic acid, a key component of specific proteoglycans and glycolipids. Consistent with being loss-of-function alleles, we show using patients’ primary fibroblasts and biochemical assays, that these mutations either impair UGDH stability, oligomerization, or enzymatic activity. In vitro, patient-derived cerebral organoids are smaller with a reduced number of proliferating neuronal progenitors while mutant ugdh zebrafish do not phenocopy the human disease. Our study defines UGDH as a key player for the production of extracellular matrix components that are essential for human brain development. Based on the incidence of variants observed, UGDH mutations are likely to be a frequent cause of recessive epileptic encephalopathy., German Research Foundation (DFG); European Union (European Union); NEUROMICS Network; International Coordination Action (ICA); Fund for Scientific Research Flanders (FWO); Netherlands Organization for Scientific Research (ZONMW VIDI); National Medical Research Council, Singapore; A Strategic Positioning Fund on Genetic Orphan Diseases (GODAFIT); Industry Alignment Fund on Singapore Childhood Undiagnosed Diseases Program (SUREKids); Biomedical Research Council, A*STAR; Diana and Steve Marienhoff Fashion Industries Guild Endowed Fellowship in Pediatric Neuromuscular Diseases; Fondazione Bambino Gesù (Vite Coraggiose); Canadian Institutes of Health Research; Natural Sciences and Engineering Research Council of Canada; Eurocores Program EuroEPINOMICS; University of Antwerp Research Fund; FRAXA Foundation; Brain & Behavior Research Foundation, NARSAD Young Investigator Grant
- Published
- 2020
13. Two zebrafish cacna1s loss-of-function variants provide models of mild and severe CACNA1S-related myopathy.
- Author
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Endo Y, Groom L, Wang SM, Pannia E, Griffiths NW, Van Gennip JLM, Ciruna B, Laporte J, Dirksen RT, and Dowling JJ
- Subjects
- Animals, Humans, Muscle, Skeletal metabolism, Mutation, Calcium Channels, L-Type genetics, Calcium Channels, L-Type metabolism, Muscular Diseases pathology, Zebrafish genetics, Zebrafish metabolism, Zebrafish Proteins metabolism
- Abstract
CACNA1S-related myopathy, due to pathogenic variants in the CACNA1S gene, is a recently described congenital muscle disease. Disease associated variants result in loss of gene expression and/or reduction of Cav1.1 protein stability. There is an incomplete understanding of the underlying disease pathomechanisms and no effective therapies are currently available. A barrier to the study of this myopathy is the lack of a suitable animal model that phenocopies key aspects of the disease. To address this barrier, we generated knockouts of the two zebrafish CACNA1S paralogs, cacna1sa and cacna1sb. Double knockout fish exhibit severe weakness and early death, and are characterized by the absence of Cav1.1 α1 subunit expression, abnormal triad structure, and impaired excitation-contraction coupling, thus mirroring the severe form of human CACNA1S-related myopathy. A double mutant (cacna1sa homozygous, cacna1sb heterozygote) exhibits normal development, but displays reduced body size, abnormal facial structure, and cores on muscle pathologic examination, thus phenocopying the mild form of human CACNA1S-related myopathy. In summary, we generated and characterized the first cacna1s zebrafish loss-of-function mutants, and show them to be faithful models of severe and mild forms of human CACNA1S-related myopathy suitable for future mechanistic studies and therapy development., (© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)
- Published
- 2024
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14. Apollo-NADP + reveals in vivo adaptation of NADPH/NADP + metabolism in electrically activated pancreatic β cells.
- Author
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Bui CV, Boswell CW, Ciruna B, and Rocheleau JV
- Subjects
- Animals, NADP metabolism, Zebrafish metabolism, Oxidation-Reduction, Folic Acid metabolism, Insulin-Secreting Cells
- Abstract
Several genetically encoded sensors have been developed to study live cell NADPH/NADP
+ dynamics, but their use has been predominantly in vitro. Here, we developed an in vivo assay using the Apollo-NADP+ sensor and microfluidic devices to measure endogenous NADPH/NADP+ dynamics in the pancreatic β cells of live zebrafish embryos. Flux through the pentose phosphate pathway, the main source of NADPH in many cell types, has been reported to be low in β cells. Thus, it is unclear how these cells compensate to meet NADPH demands. Using our assay, we show that pyruvate cycling is the main source of NADP+ reduction in β cells, with contributions from folate cycling after acute electrical activation. INS1E β cells also showed a stress-induced increase in folate cycling and further suggested that this cycling requires both increased glycolytic intermediates and cytosolic NAD+ . Overall, we show in vivo application of the Apollo-NADP+ sensor and reveal that β cells are capable of adapting NADPH/NADP+ redox during stress.- Published
- 2023
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15. Loss of zebrafish pkd1l1 causes biliary defects that have implications for biliary atresia splenic malformation.
- Author
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Ali RQ, Meyer-Miner A, David-Rachel M, Lee FJH, Wilkins BJ, Karpen SJ, Ciruna B, Ghanekar A, and Kamath BM
- Subjects
- Animals, Membrane Proteins genetics, Spleen, Abnormalities, Multiple, Biliary Atresia, Biliary Tract, Zebrafish genetics
- Abstract
Biliary atresia is a fibroinflammatory neonatal disease with no effective therapies. A subset of cases (10-20%) is associated with laterality defects - labeled biliary atresia splenic malformation (BASM) syndrome. Recently, whole-exome sequencing of patients with BASM identified deleterious variants in PKD1L1. PKD1L1 is involved in left-right axis determination; however, its role in cholangiocytes is unknown. We generated the pkd1l1hsc117 allele using CRISPR/Cas9 mutagenesis in zebrafish to determine the role of Pkd1l1 in biliary development and function. Wild-type and mutant larvae were assessed for laterality defects, biliary function and biliary tree architecture at 5 days post fertilization. pkd1l1hsc117 mutant larvae exhibited early left-right patterning defects. The gallbladder was positioned on the left in 47% of mutants compared to 4% of wild-type larvae. Accumulation of PED6 in the gallbladder, an indicator of hepatobiliary function, was significantly reduced in pkd1l1hsc117 mutants (46%) compared to wild-type larvae (4%). pkd1l1hsc117 larvae exhibited fewer biliary epithelial cells and reduced density of the intrahepatic biliary network compared to those in wild-type larvae. These data highlight the essential role of pkd1l1 in normal development and function of the zebrafish biliary system, supporting a role for this gene as a cause of BASM., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2023. Published by The Company of Biologists Ltd.)
- Published
- 2023
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16. COL11A2 as a candidate gene for vertebral malformations and congenital scoliosis.
- Author
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Rebello D, Wohler E, Erfani V, Li G, Aguilera AN, Santiago-Cornier A, Zhao S, Hwang SW, Steiner RD, Zhang TJ, Gurnett CA, Raggio C, Wu N, Sobreira N, Giampietro PF, and Ciruna B
- Subjects
- Animals, Humans, Zebrafish genetics, Spine abnormalities, Mutation, Missense, Collagen Type XI genetics, Scoliosis genetics, Abnormalities, Multiple genetics
- Abstract
Human vertebral malformations (VMs) have an estimated incidence of 1/2000 and are associated with significant health problems including congenital scoliosis (CS) and recurrent organ system malformation syndromes such as VACTERL (vertebral anomalies; anal abnormalities; cardiac abnormalities; tracheo-esophageal fistula; renal anomalies; limb anomalies). The genetic cause for the vast majority of VMs are unknown. In a CS/VM patient cohort, three COL11A2 variants (R130W, R1407L and R1413H) were identified in two patients with cervical VM. A third patient with a T9 hemivertebra and the R130W variant was identified from a separate study. These substitutions are predicted to be damaging to protein function, and R130 and R1407 residues are conserved in zebrafish Col11a2. To determine the role for COL11A2 in vertebral development, CRISPR/Cas9 was used to create a nonsense mutation (col11a2L642*) as well as a full gene locus deletion (col11a2del) in zebrafish. Both col11a2L642*/L642* and col11a2del/del mutant zebrafish exhibit vertebral fusions in the caudal spine, which form due to mineralization across intervertebral segments. To determine the functional consequence of VM-associated variants, we assayed their ability to suppress col11a2del VM phenotypes following transgenic expression within the developing spine. While wildtype col11a2 expression suppresses fusions in col11a2del/+ and col11a2del/del backgrounds, patient missense variant-bearing col11a2 failed to rescue the loss-of-function phenotype in these animals. These results highlight an essential role for COL11A2 in vertebral development and support a pathogenic role for two missense variants in CS., (© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)
- Published
- 2023
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17. Live imaging and conditional disruption of native PCP activity using endogenously tagged zebrafish sfGFP-Vangl2.
- Author
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Jussila M, Boswell CW, Griffiths NW, Pumputis PG, and Ciruna B
- Subjects
- Animals, Embryonic Development genetics, Membrane Proteins genetics, Membrane Proteins metabolism, Zebrafish Proteins genetics, Zebrafish Proteins metabolism, Cell Polarity genetics, Zebrafish
- Abstract
Tissue-wide coordination of polarized cytoskeletal organization and cell behaviour, critical for normal development, is controlled by asymmetric membrane localization of non-canonical Wnt/planar cell polarity (PCP) signalling components. Understanding the dynamic regulation of PCP thus requires visualization of these polarity proteins in vivo. Here we utilize CRISPR/Cas9 genome editing to introduce a fluorescent reporter onto the core PCP component, Vangl2, in zebrafish. Through live imaging of endogenous sfGFP-Vangl2 expression, we report on the authentic regulation of vertebrate PCP during embryogenesis. Furthermore, we couple sfGFP-Vangl2 with conditional zGrad GFP-nanobody degradation methodologies to interrogate tissue-specific functions for PCP. Remarkably, loss of Vangl2 in foxj1a-positive cell lineages causes ependymal cell cilia and Reissner fiber formation defects as well as idiopathic-like scoliosis. Together, our studies provide crucial insights into the establishment and maintenance of vertebrate PCP and create a powerful experimental paradigm for investigating post-embryonic and tissue-specific functions for Vangl2 in development and disease., (© 2022. The Author(s).)
- Published
- 2022
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18. Resolving primary pathomechanisms driving idiopathic-like spinal curvature using a new katnb1 scoliosis model.
- Author
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Meyer-Miner A, Van Gennip JLM, Henke K, Harris MP, and Ciruna B
- Abstract
Idiopathic scoliosis (IS) refers to abnormal spinal curvatures that occur in the absence of vertebral or neuromuscular defects. IS accounts for 80% of human spinal deformity, afflicts ∼3% of children worldwide, yet pathogenic mechanisms are poorly understood. A key role for cerebrospinal fluid (CSF) homeostasis in zebrafish spine development has been identified. Specifically, defects in cilia motility of brain ependymal cells (EC), CSF flow, and/or Reissner fiber (RF) assembly are observed to induce neuroinflammation, oxidative stress, abnormal CSF-contacting neuron activity, and urotensin peptide expression, all associating with scoliosis. However, the functional relevance of these observations to IS remains unclear. Here we characterize zebrafish katnb1 mutants as a new IS model. We define essential roles for Katnb1 in motile ciliated lineages, uncouple EC cilia and RF formation defects from spinal curvature, and identify abnormal CSF flow and cell stress responses as shared pathogenic signatures associated with scoliosis across diverse zebrafish models., Competing Interests: The authors declare no competing interests., (© 2022 The Author(s).)
- Published
- 2022
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19. The NEMP family supports metazoan fertility and nuclear envelope stiffness.
- Author
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Tsatskis Y, Rosenfeld R, Pearson JD, Boswell C, Qu Y, Kim K, Fabian L, Mohammad A, Wang X, Robson MI, Krchma K, Wu J, Gonçalves J, Hodzic D, Wu S, Potter D, Pelletier L, Dunham WH, Gingras AC, Sun Y, Meng J, Godt D, Schedl T, Ciruna B, Choi K, Perry JRB, Bremner R, Schirmer EC, Brill JA, Jurisicova A, and McNeill H
- Abstract
Human genome-wide association studies have linked single-nucleotide polymorphisms (SNPs) in NEMP1 ( nuclear envelope membrane protein 1 ) with early menopause; however, it is unclear whether NEMP1 has any role in fertility. We show that whole-animal loss of NEMP1 homologs in Drosophila , Caenorhabditis elegans , zebrafish, and mice leads to sterility or early loss of fertility. Loss of Nemp leads to nuclear shaping defects, most prominently in the germ line. Biochemical, biophysical, and genetic studies reveal that NEMP proteins support the mechanical stiffness of the germline nuclear envelope via formation of a NEMP-EMERIN complex. These data indicate that the germline nuclear envelope has specialized mechanical properties and that NEMP proteins play essential and conserved roles in fertility., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).)
- Published
- 2020
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20. SCO-Spondin Defects and Neuroinflammation Are Conserved Mechanisms Driving Spinal Deformity across Genetic Models of Idiopathic Scoliosis.
- Author
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Rose CD, Pompili D, Henke K, Van Gennip JLM, Meyer-Miner A, Rana R, Gobron S, Harris MP, Nitz M, and Ciruna B
- Subjects
- Animals, Cell Adhesion Molecules, Neuronal metabolism, Disease Models, Animal, Humans, Spinal Cord abnormalities, Spinal Cord growth & development, Spine abnormalities, Zebrafish growth & development, Cell Adhesion Molecules, Neuronal genetics, Cerebral Ventricles metabolism, Inflammation physiopathology, Morphogenesis, Spinal Cord immunology, Spine growth & development, Zebrafish abnormalities
- Abstract
Adolescent idiopathic scoliosis (AIS) affects 3% to 4% of children between the ages of 11 and 18 [1, 2]. This disorder, characterized by abnormal three-dimensional spinal curvatures that typically develop during periods of rapid growth, occurs in the absence of congenital vertebral malformations or neuromuscular defects [1]. Genetic heterogeneity [3] and a historical lack of appropriate animal models [4] have confounded basic understanding of AIS biology; thus, treatment options remain limited [5, 6]. Recently, genetic studies using zebrafish have linked idiopathic-like scoliosis to irregularities in motile cilia-mediated cerebrospinal fluid flow [7-9]. However, because loss of cilia motility in human primary ciliary dyskinesia patients is not fully associated with scoliosis [10, 11], other pathogenic mechanisms remain to be determined. Here, we demonstrate that zebrafish scospondin (sspo) mutants develop late-onset idiopathic-like spinal curvatures in the absence of obvious cilia motility defects. Sspo is a large secreted glycoprotein functionally associated with the subcommissural organ and Reissner's fiber [12]-ancient and enigmatic organs of the brain ventricular system reported to govern cerebrospinal fluid homeostasis [13, 14], neurogenesis [12, 15-18], and embryonic morphogenesis [19]. We demonstrate that irregular deposition of Sspo within brain ventricles is associated with idiopathic-like scoliosis across diverse genetic models. Furthermore, Sspo defects are sufficient to induce oxidative stress and neuroinflammatory responses implicated in AIS pathogenesis [9]. Through screening for chemical suppressors of sspo mutant phenotypes, we also identify potent agents capable of blocking severe juvenile spine deformity. Our work thus defines a new preclinical model of AIS and provides tools to realize novel therapeutic strategies., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2020
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21. Loss-of-function mutations in UDP-Glucose 6-Dehydrogenase cause recessive developmental epileptic encephalopathy.
- Author
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Hengel H, Bosso-Lefèvre C, Grady G, Szenker-Ravi E, Li H, Pierce S, Lebigot É, Tan TT, Eio MY, Narayanan G, Utami KH, Yau M, Handal N, Deigendesch W, Keimer R, Marzouqa HM, Gunay-Aygun M, Muriello MJ, Verhelst H, Weckhuysen S, Mahida S, Naidu S, Thomas TG, Lim JY, Tan ES, Haye D, Willemsen MAAP, Oegema R, Mitchell WG, Pierson TM, Andrews MV, Willing MC, Rodan LH, Barakat TS, van Slegtenhorst M, Gavrilova RH, Martinelli D, Gilboa T, Tamim AM, Hashem MO, AlSayed MD, Abdulrahim MM, Al-Owain M, Awaji A, Mahmoud AAH, Faqeih EA, Asmari AA, Algain SM, Jad LA, Aldhalaan HM, Helbig I, Koolen DA, Riess A, Kraegeloh-Mann I, Bauer P, Gulsuner S, Stamberger H, Ng AYJ, Tang S, Tohari S, Keren B, Schultz-Rogers LE, Klee EW, Barresi S, Tartaglia M, Mor-Shaked H, Maddirevula S, Begtrup A, Telegrafi A, Pfundt R, Schüle R, Ciruna B, Bonnard C, Pouladi MA, Stewart JC, Claridge-Chang A, Lefeber DJ, Alkuraya FS, Mathuru AS, Venkatesh B, Barycki JJ, Simpson MA, Jamuar SS, Schöls L, and Reversade B
- Subjects
- Adolescent, Alleles, Animals, Child, Child, Preschool, Female, Humans, Infant, Kinetics, Male, Organoids pathology, Oxidoreductases chemistry, Pedigree, Protein Domains, Syndrome, Zebrafish, Epilepsy genetics, Genes, Recessive, Loss of Function Mutation genetics, Oxidoreductases genetics, Uridine Diphosphate Glucose Dehydrogenase genetics
- Abstract
Developmental epileptic encephalopathies are devastating disorders characterized by intractable epileptic seizures and developmental delay. Here, we report an allelic series of germline recessive mutations in UGDH in 36 cases from 25 families presenting with epileptic encephalopathy with developmental delay and hypotonia. UGDH encodes an oxidoreductase that converts UDP-glucose to UDP-glucuronic acid, a key component of specific proteoglycans and glycolipids. Consistent with being loss-of-function alleles, we show using patients' primary fibroblasts and biochemical assays, that these mutations either impair UGDH stability, oligomerization, or enzymatic activity. In vitro, patient-derived cerebral organoids are smaller with a reduced number of proliferating neuronal progenitors while mutant ugdh zebrafish do not phenocopy the human disease. Our study defines UGDH as a key player for the production of extracellular matrix components that are essential for human brain development. Based on the incidence of variants observed, UGDH mutations are likely to be a frequent cause of recessive epileptic encephalopathy.
- Published
- 2020
- Full Text
- View/download PDF
22. Mutations in Kinesin family member 6 reveal specific role in ependymal cell ciliogenesis and human neurological development.
- Author
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Konjikusic MJ, Yeetong P, Boswell CW, Lee C, Roberson EC, Ittiwut R, Suphapeetiporn K, Ciruna B, Gurnett CA, Wallingford JB, Shotelersuk V, and Gray RS
- Subjects
- Amino Acid Sequence, Animals, Animals, Genetically Modified, Base Sequence, Child, Cilia metabolism, Cilia pathology, Consanguinity, Ependyma metabolism, Female, Gene Expression, Homozygote, Humans, Hydrocephalus genetics, Intellectual Disability genetics, Kinesins deficiency, Kinesins metabolism, Kinesins physiology, Male, Mice, Mice, Transgenic, Models, Animal, Neurodevelopmental Disorders metabolism, Neurodevelopmental Disorders pathology, Pedigree, Sequence Deletion, Tissue Distribution, Xenopus laevis, Zebrafish, Ependyma abnormalities, Kinesins genetics, Mutation, Neurodevelopmental Disorders genetics
- Abstract
Cerebrospinal fluid flow is crucial for neurodevelopment and homeostasis of the ventricular system of the brain, with localized flow being established by the polarized beating of the ependymal cell (EC) cilia. Here, we report a homozygous one base-pair deletion, c.1193delT (p.Leu398Glnfs*2), in the Kinesin Family Member 6 (KIF6) gene in a child displaying neurodevelopmental defects and intellectual disability. To test the pathogenicity of this novel human KIF6 mutation we engineered an analogous C-terminal truncating mutation in mouse. These mutant mice display severe, postnatal-onset hydrocephalus. We generated a Kif6-LacZ transgenic mouse strain and report expression specifically and uniquely within the ependymal cells (ECs) of the brain, without labeling other multiciliated mouse tissues. Analysis of Kif6 mutant mice with scanning electron microscopy (SEM) and immunofluorescence (IF) revealed specific defects in the formation of EC cilia, without obvious effect of cilia of other multiciliated tissues. Dilation of the ventricular system and defects in the formation of EC cilia were also observed in adult kif6 mutant zebrafish. Finally, we report Kif6-GFP localization at the axoneme and basal bodies of multi-ciliated cells (MCCs) of the mucociliary Xenopus epidermis. Overall, this work describes the first clinically-defined KIF6 homozygous null mutation in human and defines KIF6 as a conserved mediator of neurological development with a specific role for EC ciliogenesis in vertebrates., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2018
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23. Zebrafish models of non-canonical Wnt/planar cell polarity signalling: fishing for valuable insight into vertebrate polarized cell behavior.
- Author
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Jussila M and Ciruna B
- Subjects
- Animals, Models, Animal, Signal Transduction, Zebrafish embryology, Zebrafish genetics, Zebrafish metabolism, Cell Polarity physiology, Morphogenesis physiology, Wnt Proteins metabolism, Zebrafish Proteins metabolism
- Abstract
Planar cell polarity (PCP) coordinates the uniform orientation, structure and movement of cells within the plane of a tissue or organ system. It is beautifully illustrated in the polarized arrangement of bristles and hairs that project from specialized cell surfaces of the insect abdomen and wings, and pioneering genetic studies using the fruit fly, Drosophila melanogaster, have defined a core signalling network underlying PCP. This core PCP/non-canonical Wnt signalling pathway is evolutionarily conserved, and studies in zebrafish have helped transform our understanding of PCP from a peculiarity of polarized epithelia to a more universal cellular property that orchestrates a diverse suite of polarized cell behaviors that are required for normal vertebrate development. Furthermore, application of powerful genetics, embryonic cell-transplantation, and live-imaging capabilities afforded by the zebrafish model have yielded novel insights into the establishment and maintenance of vertebrate PCP, over the course of complex and dynamic morphogenetic events like gastrulation and neural tube morphogenesis. Although key questions regarding vertebrate PCP remain, with the emergence of new genome-editing technologies and the promise of endogenous labeling and Cre/LoxP conditional targeting strategies, zebrafish remains poised to deliver fundamental new insights into the function and molecular dynamic regulation of PCP signalling from embryonic development through to late-onset phenotypes and adult disease states. WIREs Dev Biol 2017, 6:e267. doi: 10.1002/wdev.267 For further resources related to this article, please visit the WIREs website., (© 2017 Wiley Periodicals, Inc.)
- Published
- 2017
- Full Text
- View/download PDF
24. Understanding Idiopathic Scoliosis: A New Zebrafish School of Thought.
- Author
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Boswell CW and Ciruna B
- Subjects
- Animals, Disease Models, Animal, Humans, Mutation, Scoliosis physiopathology, Scoliosis genetics, Spine physiopathology, Zebrafish genetics
- Abstract
Idiopathic scoliosis (IS) refers to a 3D rotation of the spine that occurs in the absence of underlying vertebral anomalies or obvious physiological defects. Despite affecting approximately 4% of the population, the etiology and pathogenesis of IS remain poorly understood, largely due to genetic heterogeneity and historical lack of appropriate developmental models. Recently, zebrafish has emerged as a powerful system for studying IS, owing to well-developed genetic resources and a natural susceptibility to spinal curvatures. Here, we summarize the utility of zebrafish as a genetic and biological model of IS, examine current faithful mutant IS models, and focus on their recent advances towards understanding core mechanisms governing both normal spine morphogenesis and the pathogenesis of IS-like spinal deformities., (Copyright © 2017 Elsevier Ltd. All rights reserved.)
- Published
- 2017
- Full Text
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25. Restrictions on the Importation of Zebrafish into Canada Associated with Spring Viremia of Carp Virus.
- Author
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Hanwell D, Hutchinson SA, Collymore C, Bruce AE, Louis R, Ghalami A, Allison WT, Ekker M, Eames BF, Childs S, Kurrasch DM, Gerlai R, Thiele T, Scott I, Ciruna B, Dowling JJ, McFarlane S, Huang P, Wen XY, Akimenko MA, Waskiewicz AJ, Drapeau P, Babiuk LA, Dragon D, Smida A, Buret AG, O'Grady E, Wilson J, Sowden-Plunkett L, Robertson, and Tropepe V
- Subjects
- Animals, Canada, Fish Diseases virology, Rhabdoviridae physiology, Rhabdoviridae Infections prevention & control, Rhabdoviridae Infections transmission, Rhabdoviridae Infections virology, Commerce legislation & jurisprudence, Fish Diseases prevention & control, Fish Diseases transmission, Government Regulation, Rhabdoviridae Infections veterinary, Zebrafish
- Abstract
The zebrafish model system is helping researchers improve the health and welfare of people and animals and has become indispensable for advancing biomedical research. As genetic engineering is both resource intensive and time-consuming, sharing successfully developed genetically modified zebrafish lines throughout the international community is critical to research efficiency and to maximizing the millions of dollars in research funding. New restrictions on importation of zebrafish into Canada based on putative susceptibility to infection by the spring viremia of carp virus (SVCV) have been imposed on the scientific community. In this commentary, we review the disease profile of SVCV in fish, discuss the findings of the Canadian government's scientific assessment, how the interpretations of their assessment differ from that of the Canadian research community, and describe the negative impact of these regulations on the Canadian research community and public as it pertains to protecting the health of Canadians.
- Published
- 2016
- Full Text
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26. Anisotropic stress orients remodelling of mammalian limb bud ectoderm.
- Author
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Lau K, Tao H, Liu H, Wen J, Sturgeon K, Sorfazlian N, Lazic S, Burrows JT, Wong MD, Li D, Deimling S, Ciruna B, Scott I, Simmons C, Henkelman RM, Williams T, Hadjantonakis AK, Fernandez-Gonzalez R, Sun Y, and Hopyan S
- Subjects
- Actins metabolism, Animals, Anisotropy, Cell Communication, Cell Division, Cell Polarity, Ectoderm metabolism, Embryo Culture Techniques, Embryonic Stem Cells physiology, Feedback, Gene Expression Regulation, Developmental, Genotype, Limb Buds metabolism, Mice, Inbred C57BL, Mice, Transgenic, Microscopy, Video, Models, Biological, Morphogenesis, Phenotype, Receptor, Fibroblast Growth Factor, Type 2 genetics, Receptor, Fibroblast Growth Factor, Type 2 metabolism, Stress, Mechanical, Time Factors, beta Catenin genetics, beta Catenin metabolism, Ectoderm physiology, Limb Buds physiology, Mechanotransduction, Cellular
- Abstract
The physical forces that drive morphogenesis are not well characterized in vivo, especially among vertebrates. In the early limb bud, dorsal and ventral ectoderm converge to form the apical ectodermal ridge (AER), although the underlying mechanisms are unclear. By live imaging mouse embryos, we show that prospective AER progenitors intercalate at the dorsoventral boundary and that ectoderm remodels by concomitant cell division and neighbour exchange. Mesodermal expansion and ectodermal tension together generate a dorsoventrally biased stress pattern that orients ectodermal remodelling. Polarized distribution of cortical actin reflects this stress pattern in a β-catenin- and Fgfr2-dependent manner. Intercalation of AER progenitors generates a tensile gradient that reorients resolution of multicellular rosettes on adjacent surfaces, a process facilitated by β-catenin-dependent attachment of cortex to membrane. Therefore, feedback between tissue stress pattern and cell intercalations remodels mammalian ectoderm.
- Published
- 2015
- Full Text
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27. The clathrin adaptor AP-1 complex and Arf1 regulate planar cell polarity in vivo.
- Author
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Carvajal-Gonzalez JM, Balmer S, Mendoza M, Dussert A, Collu G, Roman AC, Weber U, Ciruna B, and Mlodzik M
- Subjects
- Adaptor Proteins, Signal Transducing metabolism, Animals, Animals, Genetically Modified, Cadherins metabolism, Carrier Proteins metabolism, DNA-Binding Proteins metabolism, Dishevelled Proteins, Drosophila metabolism, Drosophila Proteins metabolism, Early Growth Response Protein 2 genetics, Early Growth Response Protein 2 metabolism, Frizzled Receptors metabolism, Gene Expression Regulation, Developmental, LIM Domain Proteins metabolism, Membrane Proteins metabolism, MyoD Protein genetics, MyoD Protein metabolism, Phosphoproteins metabolism, Zebrafish metabolism, Zebrafish Proteins metabolism, ADP-Ribosylation Factor 1 genetics, Adaptor Protein Complex 1 genetics, Cell Polarity genetics, Drosophila embryology, Drosophila Proteins genetics, Wings, Animal embryology, Zebrafish embryology, Zebrafish Proteins genetics
- Abstract
A key step in generating planar cell polarity (PCP) is the formation of restricted junctional domains containing Frizzled/Dishevelled/Diego (Fz/Dsh/Dgo) or Van Gogh/Prickle (Vang/Pk) complexes within the same cell, stabilized via Flamingo (Fmi) across cell membranes. Although models have been proposed for how these complexes acquire and maintain their polarized localization, the machinery involved in moving core PCP proteins around cells remains unknown. We describe the AP-1 adaptor complex and Arf1 as major regulators of PCP protein trafficking in vivo. AP-1 and Arf1 disruption affects the accumulation of Fz/Fmi and Vang/Fmi complexes in the proximo-distal axis, producing severe PCP phenotypes. Using novel tools, we demonstrate a direct and specific Arf1 involvement in Fz trafficking in vivo. Moreover, we uncover a conserved Arf1 PCP function in vertebrates. Our data support a model whereby the trafficking machinery plays an important part during PCP establishment, promoting formation of polarized PCP-core complexes in vivo.
- Published
- 2015
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28. ptk7 mutant zebrafish models of congenital and idiopathic scoliosis implicate dysregulated Wnt signalling in disease.
- Author
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Hayes M, Gao X, Yu LX, Paria N, Henkelman RM, Wise CA, and Ciruna B
- Subjects
- Adolescent, Animals, Cell Adhesion Molecules metabolism, Embryo, Nonmammalian, Female, Gene Expression Regulation, Developmental, Humans, Inheritance Patterns, Male, Mutation, Receptor Protein-Tyrosine Kinases metabolism, Scoliosis congenital, Scoliosis metabolism, Scoliosis pathology, Signal Transduction, Spine growth & development, Spine metabolism, Spine pathology, Wnt Proteins genetics, Wnt Proteins metabolism, Zebrafish growth & development, Zebrafish metabolism, Zebrafish Proteins, Cell Adhesion Molecules genetics, Disease Models, Animal, Receptor Protein-Tyrosine Kinases genetics, Scoliosis genetics, Zebrafish genetics
- Abstract
Scoliosis is a complex genetic disorder of the musculoskeletal system, characterized by three-dimensional rotation of the spine. Curvatures caused by malformed vertebrae (congenital scoliosis (CS)) are apparent at birth. Spinal curvatures with no underlying vertebral abnormality (idiopathic scoliosis (IS)) most commonly manifest during adolescence. The genetic and biological mechanisms responsible for IS remain poorly understood due largely to limited experimental models. Here we describe zygotic ptk7 (Zptk7) mutant zebrafish, deficient in a critical regulator of Wnt signalling, as the first genetically defined developmental model of IS. We identify a novel sequence variant within a single IS patient that disrupts PTK7 function, consistent with a role for dysregulated Wnt activity in disease pathogenesis. Furthermore, we demonstrate that embryonic loss-of-gene function in maternal-zygotic ptk7 mutants (MZptk7) leads to vertebral anomalies associated with CS. Our data suggest novel molecular origins of, and genetic links between, congenital and idiopathic forms of disease.
- Published
- 2014
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29. Analysis of maternal-zygotic ugdh mutants reveals divergent roles for HSPGs in vertebrate embryogenesis and provides new insight into the initiation of left-right asymmetry.
- Author
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Superina S, Borovina A, and Ciruna B
- Subjects
- Animals, Drosophila embryology, Drosophila genetics, Drosophila metabolism, Drosophila Proteins metabolism, Fibroblast Growth Factors metabolism, Gastrulation genetics, Gene Expression Regulation, Developmental, Glycosaminoglycans metabolism, Hedgehog Proteins metabolism, Heparan Sulfate Proteoglycans genetics, Mice, Signal Transduction genetics, Transforming Growth Factor beta metabolism, Wnt Proteins metabolism, Zebrafish embryology, Zebrafish genetics, Zebrafish metabolism, Body Patterning genetics, Embryonic Development genetics, Heparan Sulfate Proteoglycans metabolism, Heterotaxy Syndrome genetics, Proteoglycans metabolism, Uridine Diphosphate Glucose Dehydrogenase genetics, Zygote
- Abstract
Growth factors and morphogens regulate embryonic patterning, cell fate specification, cell migration, and morphogenesis. The activity and behavior of these signaling molecules are regulated in the extracellular space through interactions with proteoglycans (Bernfield et al., 1999; Perrimon and Bernfield 2000; Lander and Selleck 2000; Selleck 2000). Proteoglycans are high molecular-weight proteins consisting of a core protein with covalently linked glycosaminoglycan (GAG) side chains, which are thought to mediate ligand interaction. Drosophila mutant embryos deficient for UDP-glucose dehydrogenase activity (Ugdh, required for GAG synthesis) exhibit abnormal Fgf, Wnt and TGFß signaling and die during gastrulation, indicating a broad and critical role for proteoglycans during early embryonic development (Lin et al., 1999; Lin and Perrimon 2000) (Hacker et al., 1997). Mouse Ugdh mutants also die at gastrulation, however, only Fgf signaling appears disrupted (Garcia-Garcia and Anderson, 2003). These findings suggested a possible divergence in the requirement for proteoglycans during Drosophila and mouse embryogenesis, and that mammals may have evolved alternative means of regulating Wnt and TGFß activity. To further examine the function of proteoglycans in vertebrate development, we have characterized zebrafish mutants devoid of both maternal and zygotic Ugdh/Jekyll activity (MZjekyll). We demonstrate that MZjekyll mutant embryos display abnormal Fgf, Shh, and Wnt signaling activities, with concomitant defects in central nervous system patterning, cardiac ventricular fate specification and axial morphogenesis. Furthermore, we uncover a novel role for proteoglycans in left-right pattern formation. Our findings resolve longstanding questions into the evolutionary conservation of Ugdh function and provide new mechanistic insights into the initiation of left-right asymmetry., (Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2014
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30. IFT88 plays a cilia- and PCP-independent role in controlling oriented cell divisions during vertebrate embryonic development.
- Author
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Borovina A and Ciruna B
- Subjects
- Animals, Animals, Genetically Modified, Cell Division physiology, Cell Polarity physiology, Female, Male, Adaptor Proteins, Signal Transducing physiology, Cilia physiology, Embryonic Development physiology, Zebrafish embryology, Zebrafish Proteins physiology
- Abstract
The role for cilia in establishing planar cell polarity (PCP) is contentious. Although knockdown of genes known to function in ciliogenesis has been reported to cause PCP-related morphogenesis defects in zebrafish, genetic mutations affecting intraflagellar transport (IFT) do not show PCP phenotypes despite the requirement for IFT in cilia formation. This discrepancy has been attributed to off-target effects of antisense morpholino oligonucleotide (MO) injection, confounding maternal effects in zygotic mutant embryos, or an inability to distinguish between cilia-dependent versus cilia-independent protein functions. To determine the role of cilia in PCP, we generated maternal + zygotic IFT88 (MZift88) mutant zebrafish embryos, which never form cilia. We clearly demonstrate that cilia are not required to establish PCP. Rather, IFT88 plays a cilia-independent role in controlling oriented cell divisions at gastrulation and neurulation. Our results have important implications for the interpretation of cilia gene function in normal development and in disease., (Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2013
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31. Zebrabow: multispectral cell labeling for cell tracing and lineage analysis in zebrafish.
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Pan YA, Freundlich T, Weissman TA, Schoppik D, Wang XC, Zimmerman S, Ciruna B, Sanes JR, Lichtman JW, and Schier AF
- Subjects
- Animals, Animals, Genetically Modified embryology, Animals, Genetically Modified metabolism, Cell Lineage, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, Integrases genetics, Integrases metabolism, Zebrafish metabolism, Zebrafish embryology
- Abstract
Advances in imaging and cell-labeling techniques have greatly enhanced our understanding of developmental and neurobiological processes. Among vertebrates, zebrafish is uniquely suited for in vivo imaging owing to its small size and optical translucency. However, distinguishing and following cells over extended time periods remains difficult. Previous studies have demonstrated that Cre recombinase-mediated recombination can lead to combinatorial expression of spectrally distinct fluorescent proteins (RFP, YFP and CFP) in neighboring cells, creating a 'Brainbow' of colors. The random combination of fluorescent proteins provides a way to distinguish adjacent cells, visualize cellular interactions and perform lineage analyses. Here, we describe Zebrabow (Zebrafish Brainbow) tools for in vivo multicolor imaging in zebrafish. First, we show that the broadly expressed ubi:Zebrabow line provides diverse color profiles that can be optimized by modulating Cre activity. Second, we find that colors are inherited equally among daughter cells and remain stable throughout embryonic and larval stages. Third, we show that UAS:Zebrabow lines can be used in combination with Gal4 to generate broad or tissue-specific expression patterns and facilitate tracing of axonal processes. Fourth, we demonstrate that Zebrabow can be used for long-term lineage analysis. Using the cornea as a model system, we provide evidence that embryonic corneal epithelial clones are replaced by large, wedge-shaped clones formed by centripetal expansion of cells from the peripheral cornea. The Zebrabow tool set presented here provides a resource for next-generation color-based anatomical and lineage analyses in zebrafish.
- Published
- 2013
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32. Ptk7 promotes non-canonical Wnt/PCP-mediated morphogenesis and inhibits Wnt/β-catenin-dependent cell fate decisions during vertebrate development.
- Author
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Hayes M, Naito M, Daulat A, Angers S, and Ciruna B
- Subjects
- Animals, Cell Polarity physiology, DNA Primers genetics, Gene Expression Regulation, Developmental physiology, Gene Targeting, HEK293 Cells, Humans, In Situ Hybridization, Microscopy, Confocal, Mutagenesis, Neural Tube embryology, Receptor Protein-Tyrosine Kinases genetics, Reverse Transcriptase Polymerase Chain Reaction, Zebrafish Proteins, Cell Differentiation physiology, Morphogenesis physiology, Protein-Tyrosine Kinases metabolism, Receptor Protein-Tyrosine Kinases metabolism, Wnt Signaling Pathway physiology, Zebrafish embryology, beta Catenin metabolism
- Abstract
Using zebrafish, we have characterised the function of Protein tyrosine kinase 7 (Ptk7), a transmembrane pseudokinase implicated in Wnt signal transduction during embryonic development and in cancer. Ptk7 is a known regulator of mammalian neural tube closure and Xenopus convergent extension movement. However, conflicting reports have indicated both positive and negative roles for Ptk7 in canonical Wnt/β-catenin signalling. To clarify the function of Ptk7 in vertebrate embryonic patterning and morphogenesis, we generated maternal-zygotic (MZ) ptk7 mutant zebrafish using a zinc-finger nuclease (ZFN) gene targeting approach. Early loss of zebrafish Ptk7 leads to defects in axial convergence and extension, neural tube morphogenesis and loss of planar cell polarity (PCP). Furthermore, during late gastrula and segmentation stages, we observe significant upregulation of β-catenin target gene expression and demonstrate a clear role for Ptk7 in attenuating canonical Wnt/β-catenin activity in vivo. MZptk7 mutants display expanded differentiation of paraxial mesoderm within the tailbud, suggesting an important role for Ptk7 in regulating canonical Wnt-dependent fate specification within posterior stem cell pools post-gastrulation. Furthermore, we demonstrate that a plasma membrane-tethered Ptk7 extracellular fragment is sufficient to rescue both PCP morphogenesis and Wnt/β-catenin patterning defects in MZptk7 mutant embryos. Our results indicate that the extracellular domain of Ptk7 acts as an important regulator of both non-canonical Wnt/PCP and canonical Wnt/β-catenin signalling in multiple vertebrate developmental contexts, with important implications for the upregulated PTK7 expression observed in human cancers.
- Published
- 2013
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33. Variation of BMP3 contributes to dog breed skull diversity.
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Schoenebeck JJ, Hutchinson SA, Byers A, Beale HC, Carrington B, Faden DL, Rimbault M, Decker B, Kidd JM, Sood R, Boyko AR, Fondon JW 3rd, Wayne RK, Bustamante CD, Ciruna B, and Ostrander EA
- Subjects
- Animals, Biological Evolution, Breeding, Chromosome Mapping, Genome-Wide Association Study, Genotype, Humans, Mutation, Missense, Pets, Phenotype, Skull anatomy & histology, Zebrafish genetics, Bone Morphogenetic Protein 3 genetics, Craniosynostoses genetics, Dogs genetics, Genetic Variation, Quantitative Trait Loci, Skull metabolism
- Abstract
Since the beginnings of domestication, the craniofacial architecture of the domestic dog has morphed and radiated to human whims. By beginning to define the genetic underpinnings of breed skull shapes, we can elucidate mechanisms of morphological diversification while presenting a framework for understanding human cephalic disorders. Using intrabreed association mapping with museum specimen measurements, we show that skull shape is regulated by at least five quantitative trait loci (QTLs). Our detailed analysis using whole-genome sequencing uncovers a missense mutation in BMP3. Validation studies in zebrafish show that Bmp3 function in cranial development is ancient. Our study reveals the causal variant for a canine QTL contributing to a major morphologic trait., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2012
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34. Nedd4-1 binds and ubiquitylates activated FGFR1 to control its endocytosis and function.
- Author
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Persaud A, Alberts P, Hayes M, Guettler S, Clarke I, Sicheri F, Dirks P, Ciruna B, and Rotin D
- Subjects
- Amino Acid Motifs, Amino Acid Sequence, Animals, Binding Sites, Body Patterning physiology, Cell Differentiation physiology, Central Nervous System embryology, Endosomal Sorting Complexes Required for Transport genetics, Gene Knockdown Techniques, Humans, Molecular Sequence Data, Nedd4 Ubiquitin Protein Ligases, Neurons cytology, Peptide Fragments metabolism, Protein Binding, Protein Interaction Mapping, Protein Transport, Rats, Receptor, Fibroblast Growth Factor, Type 1 chemistry, Receptor, Fibroblast Growth Factor, Type 1 genetics, Recombinant Fusion Proteins physiology, Signal Transduction physiology, Species Specificity, Stem Cells cytology, Substrate Specificity, Ubiquitin-Protein Ligases genetics, Ubiquitination, Zebrafish embryology, Endocytosis physiology, Endosomal Sorting Complexes Required for Transport physiology, Protein Processing, Post-Translational, Receptor, Fibroblast Growth Factor, Type 1 physiology, Ubiquitin-Protein Ligases physiology
- Abstract
Fibroblast growth factor receptor 1 (FGFR1) has critical roles in cellular proliferation and differentiation during animal development and adult homeostasis. Here, we show that human Nedd4 (Nedd4-1), an E3 ubiquitin ligase comprised of a C2 domain, 4 WW domains, and a Hect domain, regulates endocytosis and signalling of FGFR1. Nedd4-1 binds directly to and ubiquitylates activated FGFR1, by interacting primarily via its WW3 domain with a novel non-canonical sequence (non-PY motif) on FGFR1. Deletion of this recognition motif (FGFR1-Δ6) abolishes Nedd4-1 binding and receptor ubiquitylation, and impairs endocytosis of activated receptor, as also observed upon Nedd4-1 knockdown. Accordingly, FGFR1-Δ6, or Nedd4-1 knockdown, exhibits sustained FGF-dependent receptor Tyr phosphorylation and downstream signalling (activation of FRS2α, Akt, Erk1/2, and PLCγ). Expression of FGFR1-Δ6 in human embryonic neural stem cells strongly promotes FGF2-dependent neuronal differentiation. Furthermore, expression of this FGFR1-Δ6 mutant in zebrafish embryos disrupts anterior neuronal patterning (head development), consistent with excessive FGFR1 signalling. These results identify Nedd4-1 as a key regulator of FGFR1 endocytosis and signalling during neuronal differentiation and embryonic development.
- Published
- 2011
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35. Insufficiency of BUBR1, a mitotic spindle checkpoint regulator, causes impaired ciliogenesis in vertebrates.
- Author
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Miyamoto T, Porazinski S, Wang H, Borovina A, Ciruna B, Shimizu A, Kajii T, Kikuchi A, Furutani-Seiki M, and Matsuura S
- Subjects
- Adaptor Proteins, Signal Transducing genetics, Adaptor Proteins, Signal Transducing metabolism, Anaphase-Promoting Complex-Cyclosome, Animals, Cdc20 Proteins, Cell Cycle Proteins metabolism, Cell Line, Cerebellum metabolism, Cerebellum pathology, Chromosome Disorders genetics, Chromosome Disorders physiopathology, Cytoplasmic Structures metabolism, Dishevelled Proteins, Dogs, Gene Knockdown Techniques, Humans, Mice, Models, Biological, Mosaicism, Oryzias, Phosphoproteins genetics, Phosphoproteins metabolism, Protein Binding, Protein Serine-Threonine Kinases deficiency, Protein Serine-Threonine Kinases genetics, Protein Stability, Signal Transduction, Ubiquitin-Protein Ligase Complexes metabolism, Vertebrates genetics, Vertebrates metabolism, Wnt Proteins metabolism, Cilia metabolism, Cilia pathology, Cytoplasmic Structures pathology, Protein Serine-Threonine Kinases metabolism
- Abstract
Budding uninhibited by benzimidazole-related 1 (BUBR1) is a central molecule of the spindle assembly checkpoint. Germline mutations in the budding uninhibited by benzimidazoles 1 homolog beta gene encoding BUBR1 cause premature chromatid separation (mosaic variegated aneuploidy) [PCS (MVA)] syndrome, which is characterized by constitutional aneuploidy and a high risk of childhood cancer. Patients with the syndrome often develop Dandy-Walker complex and polycystic kidneys; implying a critical role of BUBR1 in morphogenesis. However, little is known about the function of BUBR1 other than mitotic control. Here, we report that BUBR1 is essential for the primary cilium formation, and that the PCS (MVA) syndrome is thus a novel ciliopathy. Morpholino knockdown of bubr1 in medaka fish also caused ciliary dysfunction characterized by defects in cerebellar development and perturbed left-right asymmetry of the embryo. Biochemical analyses demonstrated that BUBR1 is required for ubiquitin-mediated proteasomal degradation of cell division cycle protein 20 in the G0 phase and maintains anaphase-promoting complex/cyclosome-CDC20 homolog 1 activity that regulates the optimal level of dishevelled for ciliogenesis.
- Published
- 2011
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- View/download PDF
36. Vangl2 directs the posterior tilting and asymmetric localization of motile primary cilia.
- Author
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Borovina A, Superina S, Voskas D, and Ciruna B
- Subjects
- ADP-Ribosylation Factors metabolism, Animals, Animals, Genetically Modified, Cilia metabolism, Embryo, Nonmammalian metabolism, Gene Expression Regulation, Developmental, Gene Knockout Techniques, Heparan Sulfate Proteoglycans metabolism, Membrane Proteins deficiency, Membrane Proteins genetics, Mice, Motion, Recombinant Fusion Proteins metabolism, Zebrafish embryology, Zebrafish genetics, Zebrafish Proteins deficiency, Zebrafish Proteins genetics, Cell Polarity, Membrane Proteins metabolism, Neuroepithelial Cells metabolism, Signal Transduction genetics, Zebrafish Proteins metabolism
- Abstract
Cilia are microtubule-based organelles that project into the extracellular space, function in the perception and integration of environmental cues, and regulate Hedgehog signal transduction. The emergent association of ciliary defects with diverse and pleiotropic human disorders has fuelled investigations into the molecular genetic regulation of ciliogenesis. Although recent studies implicate planar cell polarity (PCP) in cilia formation, this conclusion is based on analyses of proteins that are not specific to, or downstream effectors of PCP signal transduction. Here we characterize zebrafish embryos devoid of all Vangl2 function, a core and specific component of the PCP signalling pathway. Using Arl13b-GFP as a live marker of the ciliary axoneme, we demonstrate that Vangl2 is not required for ciliogenesis. Instead, Vangl2 controls the posterior tilting of primary motile cilia lining the neurocoel, Kupffer's vesicle and pronephric duct. Furthermore, we show that Vangl2 is required for asymmetric localization of cilia to the posterior apical membrane of neuroepithelial cells. Our results indicate a broad and essential role for PCP in the asymmetric localization and orientation of motile primary cilia, establishing directional fluid flow implicated in normal embryonic development and disease.
- Published
- 2010
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37. Regulation of vertebrate nervous system alternative splicing and development by an SR-related protein.
- Author
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Calarco JA, Superina S, O'Hanlon D, Gabut M, Raj B, Pan Q, Skalska U, Clarke L, Gelinas D, van der Kooy D, Zhen M, Ciruna B, and Blencowe BJ
- Subjects
- Animals, Brain cytology, Cell Differentiation, Cell Line, Humans, Mice, Nerve Tissue Proteins chemistry, Nerve Tissue Proteins genetics, Neurons cytology, Nuclear Proteins chemistry, RNA-Binding Proteins chemistry, Serine-Arginine Splicing Factors, Alternative Splicing, Nerve Tissue Proteins metabolism, Nuclear Proteins metabolism, RNA-Binding Proteins metabolism
- Abstract
Alternative splicing is a key process underlying the evolution of increased proteomic and functional complexity and is especially prevalent in the mammalian nervous system. However, the factors and mechanisms governing nervous system-specific alternative splicing are not well understood. Through a genome-wide computational and expression profiling strategy, we have identified a tissue- and vertebrate-restricted Ser/Arg (SR) repeat splicing factor, the neural-specific SR-related protein of 100 kDa (nSR100). We show that nSR100 regulates an extensive network of brain-specific alternative exons enriched in genes that function in neural cell differentiation. nSR100 acts by increasing the levels of the neural/brain-enriched polypyrimidine tract binding protein and by interacting with its target transcripts. Disruption of nSR100 prevents neural cell differentiation in cell culture and in the developing zebrafish. Our results thus reveal a critical neural-specific alternative splicing regulator, the evolution of which has contributed to increased complexity in the vertebrate nervous system.
- Published
- 2009
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- View/download PDF
38. Convergence and extension movements during vertebrate gastrulation.
- Author
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Yin C, Ciruna B, and Solnica-Krezel L
- Subjects
- Animals, Cell Polarity, Mesoderm cytology, Time Factors, Cell Movement, Gastrulation physiology, Vertebrates embryology
- Abstract
During vertebrate gastrulation, coordinated cell movements shape the basic body plan. Key components of gastrulation are convergence and extension (C&E) movements, which narrow and lengthen the embryonic tissues, respectively. The rates of C&E movements differ significantly according to the position and the stage of gastrulation. Here, we review the distinct cellular behaviors that define the spatial and temporal patterns of C&E movements, with the special emphasis on zebrafish. We also summarize the molecular regulation of these cellular behaviors and the interplay between different signaling pathways that drive C&E. Finally, to ensure efficient C&E movements, cells must achieve mediolaterally-elongated cell morphology and polarize motile protrusions. We discuss the recent discoveries on the molecular and cellular mechanisms by which the mediolateral cell polarity is established.
- Published
- 2009
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39. An eosinophil immune response characterizes the inflammatory skin disease observed in Tie-2 transgenic mice.
- Author
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Voskas D, Babichev Y, Ling LS, Alami J, Shaked Y, Kerbel RS, Ciruna B, and Dumont DJ
- Subjects
- Animals, CD3 Complex immunology, Cell Count, Cytokines blood, Cytokines genetics, Flow Cytometry, Gene Expression Regulation, Mice, Mice, Transgenic, RNA, Messenger genetics, RNA, Messenger metabolism, Skin Diseases pathology, Splenomegaly immunology, Splenomegaly pathology, Th1 Cells immunology, Eosinophils immunology, Inflammation immunology, Receptor, TIE-2 metabolism, Skin Diseases immunology
- Abstract
Although mouse models of inflammatory skin diseases such as psoriasis and atopic dermatitis fail to completely phenocopy disease in humans, they provide invaluable tools to examine the molecular and cellular mechanisms responsible for the epidermal hyperplasia, inflammation, and excess angiogenesis observed in human disease. We have previously characterized a tyrosine kinase with immunoglobin-like and epidermal growth factor-like domain-2 (Tie-2) transgenic mouse model of an inflammatory skin disease exhibiting these features. More specifically, we demonstrated that the inflammatory component consisted of increased infiltration of CD3-positive T lymphocytes and mast cells in the skin. Here, we further characterize the inflammatory component in the blood and skin of Tie-2 transgenic mice at cellular and molecular levels. We observed increased numbers of CD3-positive T lymphocytes in the blood and increased infiltration of eosinophils in the skin. Furthermore, we characterized cytokine protein and gene expression in the blood and skin, respectively, and observed the deregulated expression of cytokines associated with Th1 and eosinophil immune responses. Interestingly, treatment of Tie-2 transgenic mice with anti-CD4 antibody appeared to resolve aspects of inflammation but did not resolve epidermal hyperplasia, suggesting an important role for eosinophils in mediating the inflammatory skin disease observed in Tie-2 transgenic mice.
- Published
- 2008
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- View/download PDF
40. Maternal nodal and zebrafish embryogenesis.
- Author
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Bennett JT, Stickney HL, Choi WY, Ciruna B, Talbot WS, and Schier AF
- Subjects
- Animals, Embryo, Nonmammalian metabolism, Female, Intracellular Signaling Peptides and Proteins genetics, Models, Biological, Mothers, Nodal Signaling Ligands, Ovary metabolism, Ovum metabolism, RNA Splicing, Reproducibility of Results, Zebrafish genetics, Zebrafish metabolism, Zebrafish Proteins genetics, Body Patterning, Intracellular Signaling Peptides and Proteins metabolism, Zebrafish embryology, Zebrafish Proteins metabolism
- Abstract
In fish and amphibians, the dorsal axis is specified by the asymmetric localization of maternally provided components of the Wnt signalling pathway. Gore et al. suggest that the Nodal signal Squint (Sqt) is required as a maternally provided dorsal determinant in zebrafish. Here we test their proposal and show that the maternal activities of sqt and the related Nodal gene cyclops (cyc) are not required for dorsoventral patterning.
- Published
- 2007
- Full Text
- View/download PDF
41. A fully automated robotic system for microinjection of zebrafish embryos.
- Author
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Wang W, Liu X, Gelinas D, Ciruna B, and Sun Y
- Subjects
- Animals, Animals, Genetically Modified, Base Sequence, DNA Primers, Automation, Embryo, Nonmammalian, Microinjections, Robotics, Zebrafish embryology
- Abstract
As an important embodiment of biomanipulation, injection of foreign materials (e.g., DNA, RNAi, sperm, protein, and drug compounds) into individual cells has significant implications in genetics, transgenics, assisted reproduction, and drug discovery. This paper presents a microrobotic system for fully automated zebrafish embryo injection, which overcomes the problems inherent in manual operation, such as human fatigue and large variations in success rates due to poor reproducibility. Based on computer vision and motion control, the microrobotic system performs injection at a speed of 15 zebrafish embryos (chorion unremoved) per minute, with a survival rate of 98% (n = 350 embryos), a success rate of 99% (n = 350 embryos), and a phenotypic rate of 98.5% (n = 210 embryos). The sample immobilization technique and microrobotic control method are applicable to other biological injection applications such as the injection of mouse oocytes/embryos and Drosophila embryos to enable high-throughput biological and pharmaceutical research.
- Published
- 2007
- Full Text
- View/download PDF
42. Planar cell polarity signalling couples cell division and morphogenesis during neurulation.
- Author
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Ciruna B, Jenny A, Lee D, Mlodzik M, and Schier AF
- Subjects
- Animals, Cell Division, Membrane Proteins genetics, Membrane Proteins metabolism, Mutation genetics, Zebrafish genetics, Zebrafish Proteins genetics, Zebrafish Proteins metabolism, Cell Polarity, Morphogenesis, Nervous System cytology, Nervous System embryology, Neurons cytology, Signal Transduction, Zebrafish embryology
- Abstract
Environmental and genetic aberrations lead to neural tube closure defects (NTDs) in 1 out of every 1,000 births. Mouse and frog models for these birth defects have indicated that Van Gogh-like 2 (Vangl2, also known as Strabismus) and other components of planar cell polarity (PCP) signalling might control neurulation by promoting the convergence of neural progenitors to the midline. Here we show a novel role for PCP signalling during neurulation in zebrafish. We demonstrate that non-canonical Wnt/PCP signalling polarizes neural progenitors along the anteroposterior axis. This polarity is transiently lost during cell division in the neural keel but is re-established as daughter cells reintegrate into the neuroepithelium. Loss of zebrafish Vangl2 (in trilobite mutants) abolishes the polarization of neural keel cells, disrupts re-intercalation of daughter cells into the neuroepithelium, and results in ectopic neural progenitor accumulations and NTDs. Remarkably, blocking cell division leads to rescue of trilobite neural tube morphogenesis despite persistent defects in convergence and extension. These results reveal a function for PCP signalling in coupling cell division and morphogenesis at neurulation and indicate a previously unrecognized mechanism that might underlie NTDs.
- Published
- 2006
- Full Text
- View/download PDF
43. Production of maternal-zygotic mutant zebrafish by germ-line replacement.
- Author
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Ciruna B, Weidinger G, Knaut H, Thisse B, Thisse C, Raz E, and Schier AF
- Subjects
- Animals, Base Sequence, Cell Transplantation, DNA Primers, Female, Genotype, Germ-Line Mutation, Male, RNA, Messenger genetics, Transplantation Chimera, Genomic Imprinting, Zebrafish genetics, Zygote physiology
- Abstract
We report a generally applicable strategy for transferring zygotic lethal mutations through the zebrafish germ line. By using a morpholino oligonucleotide that blocks primordial germ cell (PGC) development, we generate embryos devoid of endogenous PGCs to serve as hosts for the transplantation of germ cells derived from homozygous mutant donors. Successful transfers are identified by the localization of specifically labeled donor PGCs to the region of the developing gonad in chimeric embryos. This strategy, which results in the complete replacement of the host germ line with donor PGCs, was validated by the generation of maternal and maternal-zygotic mutants for the miles apart locus. This germ-line replacement technique provides a powerful tool for studying the maternal effects of zygotic lethal mutations. Furthermore, the ability to generate large clutches of purely mutant embryos will greatly facilitate embryological, genetic, genomic, and biochemical studies.
- Published
- 2002
- Full Text
- View/download PDF
44. The SH2 tyrosine phosphatase shp2 is required for mammalian limb development.
- Author
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Saxton TM, Ciruna BG, Holmyard D, Kulkarni S, Harpal K, Rossant J, and Pawson T
- Subjects
- Animals, Branchial Region cytology, Branchial Region enzymology, Cell Adhesion genetics, Cell Division genetics, Cell Movement genetics, Cell Size genetics, Chimera genetics, Ectoderm cytology, Ectoderm enzymology, Forelimb enzymology, Genes, Reporter, Hindlimb enzymology, Intracellular Signaling Peptides and Proteins, Limb Buds cytology, Limb Buds embryology, Mesoderm cytology, Mesoderm enzymology, Mice, Protein Tyrosine Phosphatase, Non-Receptor Type 1, Protein Tyrosine Phosphatase, Non-Receptor Type 11, Protein Tyrosine Phosphatase, Non-Receptor Type 6, Protein Tyrosine Phosphatases metabolism, RNA, Messenger biosynthesis, Receptor Protein-Tyrosine Kinases metabolism, Receptor, Fibroblast Growth Factor, Type 1, Receptors, Fibroblast Growth Factor metabolism, SH2 Domain-Containing Protein Tyrosine Phosphatases, Signal Transduction genetics, Stem Cells cytology, Transgenes, beta-Galactosidase genetics, Forelimb embryology, Hindlimb embryology, Limb Buds enzymology, Protein Tyrosine Phosphatases genetics, src Homology Domains genetics
- Abstract
The tyrosine phosphatase Shp2 is recruited into tyrosine-kinase signalling pathways through binding of its two amino-terminal SH2 domains to specific phosphotyrosine motifs, concurrent with its re-localization and stimulation of phosphatase activity. Shp2 can potentiate signalling through the MAP-kinase pathway and is required during early mouse development for gastrulation. Chimaeric analysis can identify, by study of phenotypically normal embryos, tissues that tolerate mutant cells (and therefore do not require the mutated gene) or lack mutant cells (and presumably require the mutated gene during their developmental history). We therefore generated chimaeric mouse embryos to explore the cellular requirements for Shp2. This analysis revealed an obligatory role for Shp2 during outgrowth of the limb. Shp2 is specifically required in mesenchyme cells of the progress zone (PZ), directly beneath the distal ectoderm of the limb bud. Comparison of Ptpn11 (encoding Shp2)-mutant and Fgfr1 (encoding fibroblast growth factor receptor-1)-mutant chimaeric limbs indicated that in both cases mutant cells fail to contribute to the PZ of phenotypically normal chimaeras, leading to the hypothesis that a signal transduction pathway, initiated by Fgfr1 and acting through Shp2, is essential within PZ cells. Rather than integrating proliferative signals, Shp2 probably exerts its effects on limb development by influencing cell shape, movement or adhesion. Furthermore, the branchial arches, which also use Fgfs during bud outgrowth, similarly require Shp2. Thus, Shp2 regulates phosphotyrosine-signalling events during the complex ectodermal-mesenchymal interactions that regulate mammalian budding morphogenesis.
- Published
- 2000
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45. Distinct neural stem cells proliferate in response to EGF and FGF in the developing mouse telencephalon.
- Author
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Tropepe V, Sibilia M, Ciruna BG, Rossant J, Wagner EF, and van der Kooy D
- Subjects
- Animals, Cell Differentiation, Cell Division drug effects, Chimera, Embryonic and Fetal Development, ErbB Receptors metabolism, Gestational Age, Immunohistochemistry, Intermediate Filament Proteins metabolism, Mice, Nestin, Ploidies, Receptors, Fibroblast Growth Factor metabolism, Signal Transduction physiology, Epidermal Growth Factor pharmacology, Fibroblast Growth Factors pharmacology, Nerve Tissue Proteins, Neurons metabolism, Stem Cells metabolism, Telencephalon embryology
- Abstract
Multipotent, self-renewing neural stem cells reside in the embryonic mouse telencephalic germinal zone. Using an in vitro neurosphere assay for neural stem cell proliferation, we demonstrate that FGF-responsive neural stem cells are present as early as E8.5 in the anterior neural plate, but EGF-responsive neural stem cells emerge later in development in a temporally and spatially specific manner. By separately blocking EGF and FGF2 signaling, we also show that EGF alone and FGF2 alone can independently elicit neural stem cell proliferation and at relatively high cell densities separate cell nonautonomous effects can substantially enhance the mitogen-induced proliferation. At lower cell densities, neural stem cell proliferation is additive in the presence of EGF and FGF2 combined, revealing two different stem cell populations. However, both FGF-responsive and EGF-responsive neural stem cells retain their self-renewal and multilineage potential, regardless of growth factor conditions. These results support a model in which separate, lineage-related EGF- and FGF-responsive neural stem cells are present in the embryonic telencephalic germinal zone., (Copyright 1999 Academic Press.)
- Published
- 1999
- Full Text
- View/download PDF
46. FGF signaling in mouse gastrulation and anteroposterior patterning.
- Author
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Rossant J, Ciruna B, and Partanen J
- Subjects
- Alleles, Animals, Chimera, Drosophila embryology, Mice, Mice, Knockout, Models, Biological, Receptor Protein-Tyrosine Kinases deficiency, Receptor Protein-Tyrosine Kinases genetics, Receptor, Fibroblast Growth Factor, Type 1, Receptors, Fibroblast Growth Factor deficiency, Receptors, Fibroblast Growth Factor genetics, Stem Cells physiology, Body Patterning, Embryonic and Fetal Development, Fibroblast Growth Factors physiology, Gastrula physiology, Signal Transduction physiology
- Published
- 1997
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