The Experts below are selected from a list of 471 Experts worldwide ranked by ideXlab platform
Gabrielle Kardon - One of the best experts on this subject based on the ideXlab platform.
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cell culture system to assay candidate genes and molecular pathways implicated in congenital diaphragmatic hernias
Developmental Biology, 2020Co-Authors: Eric L Bogenschutz, Elizabeth M Sefton, Gabrielle KardonAbstract:The mammalian muscularized diaphragm is essential for respiration and defects in the developing diaphragm cause a common and frequently lethal birth defect, congenital diaphragmatic hernia (CDH). Human genetic studies have implicated more than 150 genes and multiple molecular pathways in CDH, but few of these have been validated because of the expense and time to generate mouse mutants. The Pleuroperitoneal folds (PPFs) are transient embryonic structures in diaphragm development and defects in PPFs lead to CDH. We have developed a system to culture PPF fibroblasts from E12.5 mouse embryos and show that these fibroblasts, in contrast to the commonly used NIH 3T3 fibroblasts, maintain expression of key genes in normal diaphragm development. Using pharmacological and genetic manipulations that result in CDH in vivo, we also demonstrate that differences in proliferation provide a rapid means of distinguishing healthy and impaired PPF fibroblasts. Thus, the PPF fibroblast cell culture system is an efficient tool for assaying the functional significance of CDH candidate genes and molecular pathways and will be an important resource for elucidating the complex etiology of CDH.
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cell culture system to efficiently test candidate genes and molecular pathways implicated in congenital diaphragmatic hernias
bioRxiv, 2020Co-Authors: Eric L Bogenschutz, Elizabeth M Sefton, Gabrielle KardonAbstract:The mammalian muscularized diaphragm is essential for respiration and defects in the developing diaphragm cause a common and frequently lethal birth defect, congenital diaphragmatic hernia (CDH). Human genetic studies have implicated more than 150 genes and multiple molecular pathways in CDH, but few of these have been validated because of the expense and time to generate mouse mutants. The Pleuroperitoneal folds (PPFs) are transient embryonic structures in diaphragm development and a critical cellular source of CDH. We have developed a system to culture PPF fibroblasts from E12.5 mouse embryos and show that these fibroblasts, in contrast to the commonly used NIH 3T3 fibroblasts, maintain expression of key genes in normal diaphragm development. Using pharmacological and genetic manipulations that result in CDH in vivo, we also demonstrate that differences in proliferation provide a rapid means of distinguishing healthy and impaired PPF fibroblasts. Thus, the PPF fibroblast cell culture system is an efficient tool for testing the functional significance of CDH candidate genes and molecular pathways and will be an important resource for elucidating the complex etiology of CDH.
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Muscle connective tissue controls development of the diaphragm and is a source of congenital diaphragmatic hernias
2016Co-Authors: Allyson J Merrell, Benjamin J Ellis, Zachary D Fox, Jennifer A Lawson, Jeffrey A, Gabrielle KardonAbstract:The diaphragm is an essential mammalian skeletal muscle, and defects in diaphragm development are the cause of congenital diaphragmatic hernias (CDH), a common and often lethal birth defect. The diaphragm is derived from multiple embryonic sources, but how these give rise to the diaphragm is unknown and, despite the identification of many CDH-associated genes, the etiology of CDH is incompletely understood. Using mouse genetics, we show that the Pleuroperitoneal folds (PPFs), transient embryonic structures, are the source of the diaphragm’s muscle connective tissue, regulate muscle development, and their striking migration controls diaphragm morphogenesis. Furthermore, Gata4 mosaic mutations in PPF-derived muscle connective tissue fibroblasts result in the development of localized amuscular regions that are biomechanically weaker and more compliant and lead to CDH. Thus the PPFs and muscle connective tissue are critical for diaphragm development and mutations in PPF-derived fibroblasts are a source of CDH
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muscle connective tissue controls development of the diaphragm and is a source of congenital diaphragmatic hernias
Nature Genetics, 2015Co-Authors: Allyson J Merrell, Benjamin J Ellis, Zachary D Fox, Jennifer A Lawson, Jeffrey A Weiss, Gabrielle KardonAbstract:Gabrielle Kardon and colleagues present a detailed study of diaphragm development in mice. They show that migration of connective tissue fibroblasts derived from the Pleuroperitoneal folds controls diaphragm morphogenesis and that mosaic ablation of Gata4 in this cell population results in defects resembling human congenital diaphragmatic hernias.
Robert P Lemke - One of the best experts on this subject based on the ideXlab platform.
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structure of the primordial diaphragm and defects associated with nitrofen induced cdh
Journal of Applied Physiology, 2000Co-Authors: John J Greer, David Cote, Douglas W Allan, Wei Zhang, Randal P Babiuk, Robert P Lemke, Keith BagnallAbstract:The goals of this study were to further our understanding of diaphragm embryogenesis and the pathogenesis of congenital diaphragmatic hernia (CDH). Past work suggests that the Pleuroperitoneal fold...
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recent advances in understanding the pathogenesis of nitrofen induced congenital diaphragmatic hernia
Pediatric Pulmonology, 2000Co-Authors: John J Greer, Douglas W Allan, Randal P Babiuk, Robert P LemkeAbstract:In this review, we discuss recent advances in the study of the pathogenesis of congenital diaphragmatic hernia (CDH). Much of the research has involved the use of an animal model of CDH in which diaphragmatic defects are produced in fetal rats by administering the herbicide nitrofen to dams during mid-gestation. The animal model is described and the relevance to the human condition is discussed. The data derived from the animal studies are critically assessed in the context of commonly cited hypotheses proposed for the pathogenesis of CDH. Finally, experimental strategies are proposed for systematically examining the normal and pathological formation of the Pleuroperitoneal fold. We conclude that a malformation of the primordial diaphragm, the Pleuroperitoneal fold, underlies the muscle defects associated with CDH.
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Structure of the primordial diaphragm and defects associated with nitrofen-induced CDH
2000Co-Authors: Randal P Babiuk, David Cote, Douglas W Allan, Robert P Lemke, Keith Bagnall, John J, Al P. BabiukAbstract:and Keith Bagnall. Structure of the primordial diaphragm and defects associated with nitrofen-induced CDH. J Appl Physiol 89: 2123–2129, 2000.—The goals of this study were to further our understanding of diaphragm embryogenesis and the pathogenesis of congenital diaphragmatic hernia (CDH). Past work suggests that the Pleuroperitoneal fold (PPF) is the primary source of diaphragmatic musculature. Furthermore, defects associated with an animal model of CDH can be traced back to the formation of the PPF. This study was designed to elucidate the anatomic structure of the PPF and to determine which regions of the PPF malform in the well-established nitrofen model of CDH. This was achieved by producing three-dimensional renderings con-structed from serial transverse sections of control and nitro-fen-exposed rats at embryonic day 13.5. Renderings of left
John J Greer - One of the best experts on this subject based on the ideXlab platform.
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diaphragm development and congenital diaphragmatic hernia
Seminars in Pediatric Surgery, 2007Co-Authors: Robin D Clugston, John J GreerAbstract:Advances in the understanding of normal diaphragm embryogenesis have provided the necessary foundation for novel insights into the pathogenesis of congenital diaphragmatic hernia (CDH). Although diaphragm formation is still not completely understood, we have identified key structures and periods of development that are clearly abnormal in animal models of CDH. The Pleuroperitoneal fold (PPF) is a transient structure which is the target for the neuromuscular component of the diaphragm. The PPF has been shown to be abnormal in multiple animal models of Bochdalek CDH; specifically, a malformation of the nonmuscular component of this tissue is thought to underlie the later defect in the complete diaphragm. Based on data from animal models and the examination of human postmortem tissue, we hypothesize that abnormal PPF development underlies Bochdalek CDH. Further, the understanding of the pathogenesis of rarer subtypes of CDH will be advanced by the study of various new animal models discussed in this review.
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structure of the primordial diaphragm and defects associated with nitrofen induced cdh
Journal of Applied Physiology, 2000Co-Authors: John J Greer, David Cote, Douglas W Allan, Wei Zhang, Randal P Babiuk, Robert P Lemke, Keith BagnallAbstract:The goals of this study were to further our understanding of diaphragm embryogenesis and the pathogenesis of congenital diaphragmatic hernia (CDH). Past work suggests that the Pleuroperitoneal fold...
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recent advances in understanding the pathogenesis of nitrofen induced congenital diaphragmatic hernia
Pediatric Pulmonology, 2000Co-Authors: John J Greer, Douglas W Allan, Randal P Babiuk, Robert P LemkeAbstract:In this review, we discuss recent advances in the study of the pathogenesis of congenital diaphragmatic hernia (CDH). Much of the research has involved the use of an animal model of CDH in which diaphragmatic defects are produced in fetal rats by administering the herbicide nitrofen to dams during mid-gestation. The animal model is described and the relevance to the human condition is discussed. The data derived from the animal studies are critically assessed in the context of commonly cited hypotheses proposed for the pathogenesis of CDH. Finally, experimental strategies are proposed for systematically examining the normal and pathological formation of the Pleuroperitoneal fold. We conclude that a malformation of the primordial diaphragm, the Pleuroperitoneal fold, underlies the muscle defects associated with CDH.
Alcino Lázaro Da ,silva - One of the best experts on this subject based on the ideXlab platform.
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Efeito da derivação Pleuroperitoneal no tratamento de hidrotórax recorrente induzido: pleurodese em coelhos
Sociedade Brasileira para o Desenvolvimento da Pesquisa em Cirurgia, 2003Co-Authors: Santos,edivar Pereira Dos, Meneghini,alexandre João, Alcino Lázaro Da ,silvaAbstract:OBJETIVO: Verificar a pleurodese, através da histologia hematoxilina - eosina e microscopia eletrônica, obtida após injeção de tetraciclina, em coelhos com uso de derivação Pleuroperitoneal no tratamento de hidrotórax recorrente induzido. MÉTODOS: Foram utilizados 30 coelhos New - Zealand, machos e adultos. Empregou-se anestesia dissociativa quetamina e xilazina em injeção intramuscular 0,1 cm³ para cada 100g de peso do animal. Para induzir hidrotórax utilizou-se tetraciclina 7 mg/kg, diluída em 10 cm³ de água destilada (pH 4) injetada via percutânea no 4º espaço intercostal anterior esquerdo. O ato operatório constituiu de laparotomia subcostal à esquerda com 4 cm de extensão e incisão do hemidiafragma na porção muscular de 0,5 cm de diâmetro. No grupo A (controle) realizou-se fechamento da abertura do diafragma com pontos simples e fio monofilamentar 5.0. No grupo B (derivação) após abertura do diafragma introduziu-se prótese cilíndrica com 2 cm de comprimento. No grupo C (drenagem) foi feita uma laparotomia subcostal esquerda com 2 cm de extensão e introdução de cateter nº 6. Toracotomia lateral esquerda com 2 cm de extensão e descolamento hipodérmico para aplicação do cateter pleurointercostodermoperitoneal. No pós-operatório realizou-se indução de hidrotórax repetida no 1º, 5º, 8º e 12º dia de pós-operatório nos três grupos de estudo através de injeção intrapleural de tetraciclina 7 mg/kg (pH 4) no 4º espaço intercostal anterior esquerdo. Foi feita eutanasia no 15º dia e necropsia para retirada de líquido pleural e histologia pleural, pulmonar e peritônio parietal. RESULTADOS: O acúmulo de líquido pleural resultante foi mais significante nos animais do grupo A. A irritação química resultou no espessamento pleural moderado nos grupos B e C. O índice de pleurodese foi 100% do grupo derivação Pleuroperitoneal e 80% do grupo drenagem. CONCLUSÃO: A derivação Pleuroperitoneal facilita a formação de pleurodese comprovada histologicamente, mesmo com a utilização de dose baixa de tetraciclina em coelhos
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Efeito da derivação Pleuroperitoneal no tratamento de hidrotórax recorrente induzido: pleurodese em coelhos Effect of the Pleuroperitoneal derivation in the treatment of induced recurrent hydrothorax: pleurodese in rabbits
Sociedade Brasileira para o Desenvolvimento da Pesquisa em Cirurgia, 2003Co-Authors: Edivar Pereira Dos Santos, Ademir Rocha, Alexandre João Meneghini, Alcino Lázaro Da ,silvaAbstract:OBJETIVO: Verificar a pleurodese, através da histologia hematoxilina - eosina e microscopia eletrônica, obtida após injeção de tetraciclina, em coelhos com uso de derivação Pleuroperitoneal no tratamento de hidrotórax recorrente induzido. MÉTODOS: Foram utilizados 30 coelhos New - Zealand, machos e adultos. Empregou-se anestesia dissociativa quetamina e xilazina em injeção intramuscular 0,1 cm³ para cada 100g de peso do animal. Para induzir hidrotórax utilizou-se tetraciclina 7 mg/kg, diluída em 10 cm³ de água destilada (pH 4) injetada via percutânea no 4º espaço intercostal anterior esquerdo. O ato operatório constituiu de laparotomia subcostal à esquerda com 4 cm de extensão e incisão do hemidiafragma na porção muscular de 0,5 cm de diâmetro. No grupo A (controle) realizou-se fechamento da abertura do diafragma com pontos simples e fio monofilamentar 5.0. No grupo B (derivação) após abertura do diafragma introduziu-se prótese cilíndrica com 2 cm de comprimento. No grupo C (drenagem) foi feita uma laparotomia subcostal esquerda com 2 cm de extensão e introdução de cateter nº 6. Toracotomia lateral esquerda com 2 cm de extensão e descolamento hipodérmico para aplicação do cateter pleurointercostodermoperitoneal. No pós-operatório realizou-se indução de hidrotórax repetida no 1º, 5º, 8º e 12º dia de pós-operatório nos três grupos de estudo através de injeção intrapleural de tetraciclina 7 mg/kg (pH 4) no 4º espaço intercostal anterior esquerdo. Foi feita eutanasia no 15º dia e necropsia para retirada de líquido pleural e histologia pleural, pulmonar e peritônio parietal. RESULTADOS: O acúmulo de líquido pleural resultante foi mais significante nos animais do grupo A. A irritação química resultou no espessamento pleural moderado nos grupos B e C. O índice de pleurodese foi 100% do grupo derivação Pleuroperitoneal e 80% do grupo drenagem. CONCLUSÃO: A derivação Pleuroperitoneal facilita a formação de pleurodese comprovada histologicamente, mesmo com a utilização de dose baixa de tetraciclina em coelhos.PURPOSE: In order to determine the pleurodese, after-got of tetracycline injection, the Pleuroperitoneal in the treatment of induced recurrent hydrothorax in rabbits. METHODS: Thirty male and adult New-Zealand rabbits were used. It was used dissociated anesthesia quetamina and xylazine association, (0,1 cm³ per 100g animal weight) by intramuscular injection.. In order to induce hydrothorax, 7 mg/kg of tetracycline diluted in 10 cm³ distilled water (pH 4) was percutaneous injected, in the 4th left anterior intercostal space with 25.6 mm diameter needle. The surgical procedure was a 4 cm long down rib laparatomy on the left, and a 0,5 cm diameter hemidiaphragm incision in the venter musculi; in group A, it was closure of the diaphragm opening with a single monofilament suture; in the group B, after the opening of the diaphragm, it was introduced a 2 cm long cylindrical prothesis, politetrafluor etileno (PTFE), Atrium Medical Corporation, number 3; in the group C, the surgical procedure was a 2 cm long down rib laparotomy on the left and toracotomy a 2,5 cm long on the left, and than it was introduced a catheter number 6 French (IBRAS-CBO) on the pleurointercostodermoperitoneal way. In the post operatory, it was carried out the induced and repeated hydrothorax on the 1st, 5th, 8th, and 12th days, in all groups of study, using 7 mg/kg intrapleural injection of tetracycline (pH 4,0) in the 4th left anterior intercostal space. Euthanasia took place on the 15th post operatory day, and necropsia in order to remove pleural effusion and do histological study of pleura, lung and parietal peritoneum. RESULTS: The pleural effusion resultant was more significant on group A. The pleurodese resultant index was 100% on group B, and 80% on group C. CONCLUSIONS: The Pleuroperitoneal derivation get advantage pleurodese histologic testable, again with tetracycline little dose in rabbits
Benjamin J Ellis - One of the best experts on this subject based on the ideXlab platform.
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Muscle connective tissue controls development of the diaphragm and is a source of congenital diaphragmatic hernias
2016Co-Authors: Allyson J Merrell, Benjamin J Ellis, Zachary D Fox, Jennifer A Lawson, Jeffrey A, Gabrielle KardonAbstract:The diaphragm is an essential mammalian skeletal muscle, and defects in diaphragm development are the cause of congenital diaphragmatic hernias (CDH), a common and often lethal birth defect. The diaphragm is derived from multiple embryonic sources, but how these give rise to the diaphragm is unknown and, despite the identification of many CDH-associated genes, the etiology of CDH is incompletely understood. Using mouse genetics, we show that the Pleuroperitoneal folds (PPFs), transient embryonic structures, are the source of the diaphragm’s muscle connective tissue, regulate muscle development, and their striking migration controls diaphragm morphogenesis. Furthermore, Gata4 mosaic mutations in PPF-derived muscle connective tissue fibroblasts result in the development of localized amuscular regions that are biomechanically weaker and more compliant and lead to CDH. Thus the PPFs and muscle connective tissue are critical for diaphragm development and mutations in PPF-derived fibroblasts are a source of CDH
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muscle connective tissue controls development of the diaphragm and is a source of congenital diaphragmatic hernias
Nature Genetics, 2015Co-Authors: Allyson J Merrell, Benjamin J Ellis, Zachary D Fox, Jennifer A Lawson, Jeffrey A Weiss, Gabrielle KardonAbstract:Gabrielle Kardon and colleagues present a detailed study of diaphragm development in mice. They show that migration of connective tissue fibroblasts derived from the Pleuroperitoneal folds controls diaphragm morphogenesis and that mosaic ablation of Gata4 in this cell population results in defects resembling human congenital diaphragmatic hernias.