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  1. Article ; Online: Divergent effects of calcineurin Aβ on regulatory and conventional T-cell homeostasis.

    Doetschman, Thomas / Sholl, Allyson / Chen, Hwu dau rw / Gard, Connie / Hildeman, David A / Bommireddy, Ramireddy

    Clinical immunology (Orlando, Fla.)

    2011  Volume 138, Issue 3, Page(s) 321–330

    Abstract: Calcineurin (CN) is a phosphatase that activates nuclear factor of activated T cells (NFAT). While the CN inhibitors cyclosporine A (CsA) and tacrolimus (FK506) can prevent graft rejection, they also cause inflammatory diseases. We investigated the role ... ...

    Abstract Calcineurin (CN) is a phosphatase that activates nuclear factor of activated T cells (NFAT). While the CN inhibitors cyclosporine A (CsA) and tacrolimus (FK506) can prevent graft rejection, they also cause inflammatory diseases. We investigated the role of calcineurin using mice deficient in the CN catalytic subunit Aβ (CNAβ). Cnab(-/-) mice exhibit defective thymocyte maturation, splenomegaly and hepatomegaly. Further, as Cnab(-/-) mice age, they exhibit spontaneous T-cell activation and enhanced production of proinflammatory cytokines (IL-4, IL-6, and IFNγ). FOXP3(+) T(reg) cells were significantly decreased in Cnab(-/-) mice likely contributing to increased T-cell activation. Interestingly, we found that CNAβ is critical for promotion of BCL-2 expression in FOXP3(+) T(reg) and for permitting TGFβ signaling, as TGFβ induces FOXP3 in control but not in Cnab(-/-) T-cells. Together, these data suggest that CNAβ is important for the production and maintenance of T(reg) cells and to ensure mature T-cell quiescence.
    MeSH term(s) Animals ; Calcineurin/genetics ; Calcineurin/immunology ; Cytokines/biosynthesis ; Cytokines/immunology ; Forkhead Transcription Factors/immunology ; Hepatomegaly/immunology ; Hepatomegaly/metabolism ; Homeostasis/immunology ; Lymphocyte Activation/immunology ; Mice ; Mice, Mutant Strains ; Proto-Oncogene Proteins c-bcl-2/biosynthesis ; Proto-Oncogene Proteins c-bcl-2/immunology ; Signal Transduction/immunology ; Splenomegaly/immunology ; Splenomegaly/metabolism ; T-Lymphocytes, Regulatory/immunology ; T-Lymphocytes, Regulatory/metabolism ; Transforming Growth Factor beta/immunology
    Chemical Substances Cytokines ; Forkhead Transcription Factors ; Foxp3 protein, mouse ; Proto-Oncogene Proteins c-bcl-2 ; Transforming Growth Factor beta ; Calcineurin (EC 3.1.3.16) ; protein phosphatase 3, catalytic subunit, beta isoform, mouse (EC 3.1.3.16)
    Language English
    Publishing date 2011-01-20
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 1459903-x
    ISSN 1521-7035 ; 1521-6616
    ISSN (online) 1521-7035
    ISSN 1521-6616
    DOI 10.1016/j.clim.2010.12.020
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

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  2. Article: Ligand-specific function of transforming growth factor beta in epithelial-mesenchymal transition in heart development.

    Azhar, Mohamad / Runyan, Raymond B / Gard, Connie / Sanford, L Philip / Miller, Marian L / Andringa, Anastasia / Pawlowski, Sharon / Rajan, Sudarsan / Doetschman, Thomas

    Developmental dynamics : an official publication of the American Association of Anatomists

    2009  Volume 238, Issue 2, Page(s) 431–442

    Abstract: The ligand specificity of transforming growth factor beta (TGFbeta) in vivo in mouse cardiac cushion epithelial-to-mesenchymal transition (EMT) is poorly understood. To elucidate the function of TGFbeta in cushion EMT, we analyzed Tgfb1(-/-), Tgfb2(-/-), ...

    Abstract The ligand specificity of transforming growth factor beta (TGFbeta) in vivo in mouse cardiac cushion epithelial-to-mesenchymal transition (EMT) is poorly understood. To elucidate the function of TGFbeta in cushion EMT, we analyzed Tgfb1(-/-), Tgfb2(-/-), and Tgfb3(-/-) mice between embryonic day (E) 9.5 and E14.5 using both in vitro and in vivo approaches. Atrioventricular (AV) canal collagen gel assays at E9.5 indicated normal EMT in both Tgfb1(-/-) and Tgfb3(-/-) mice. However, analysis of Tgfb2(-/-) AV explants at E9.5 and E10.5 indicated that EMT, but not cushion cell proliferation, was initially delayed but later remained persistent. This was concordant with the observation that Tgfb2(-/-) embryos, and not Tgfb1(-/-) or Tgfb3(-/-) embryos, develop enlarged cushions at E14.5 with elevated levels of well-validated indicators of EMT. Collectively, these data indicate that TGFbeta2, and not TGFbeta1 or TGFbeta3, mediates cardiac cushion EMT by promoting both the initiation and cessation of EMT.
    MeSH term(s) Animals ; Cell Differentiation/physiology ; Cell Proliferation ; Endothelial Cells/cytology ; Endothelial Cells/metabolism ; Epithelial Cells/cytology ; Epithelial Cells/physiology ; Heart/embryology ; Heart/physiology ; Ligands ; Mesoderm/cytology ; Mesoderm/embryology ; Mesoderm/metabolism ; Mice ; Mice, Knockout ; Transforming Growth Factor beta/genetics ; Transforming Growth Factor beta/physiology ; Transforming Growth Factor beta1/genetics ; Transforming Growth Factor beta1/physiology ; Transforming Growth Factor beta2/genetics ; Transforming Growth Factor beta2/physiology ; Transforming Growth Factor beta3/genetics ; Transforming Growth Factor beta3/physiology
    Chemical Substances Ligands ; Transforming Growth Factor beta ; Transforming Growth Factor beta1 ; Transforming Growth Factor beta2 ; Transforming Growth Factor beta3
    Language English
    Publishing date 2009-01-21
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 1102541-4
    ISSN 1097-0177 ; 1058-8388
    ISSN (online) 1097-0177
    ISSN 1058-8388
    DOI 10.1002/dvdy.21854
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    In stock of ZB MED Cologne/Königswinter

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  3. Article: Altered tissue behavior of a non-aneurysmal descending thoracic aorta in the mouse model of Marfan syndrome

    Haskett, Darren / Doyle, Jefferson J / Gard, Connie / Chen, Hwudaurw / Ball, Corbie / Estabrook, Mark A / Encinas, Alejandra C / Dietz, Harry C / Utzinger, Urs / Vande Geest, Jonathan P / Azhar, Mohamad

    Cell and tissue research. 2012 Jan., v. 347, no. 1

    2012  

    Abstract: Aortic aneurysm is predominantly found in the ascending aorta in patients with Marfan syndrome (MFS). However, descending aortic disease has emerged as a problem since people are living longer because of improved medical and surgical management of the ... ...

    Abstract Aortic aneurysm is predominantly found in the ascending aorta in patients with Marfan syndrome (MFS). However, descending aortic disease has emerged as a problem since people are living longer because of improved medical and surgical management of the ascending aorta. Diagnostic procedures before disease onset and the mechanisms involved in the transition of normal aortic tissue to aneurysm remain unclear. We determined signs of descending aortic disease before disease onset in mice with a mutation in the fibrillin 1 gene (Fbn1 +/C1039G), a validated mouse model of disease susceptibility and progression of aortic aneurysm of MFS. We analyzed a tubular unfixed non-aneurysmal descending thoracic aorta from 8-month-old wild-type and Fbn1 +/C1039G mice by a tubular biaxial tester that works in conjunction with a two-photon nonlinear microscope. Fbn1 +/C1039G mouse aorta was more compliant in the circumferential direction. Two-photon imaging showed defective organization of adventitial collagen fibers in the pressurized aortas of Fbn1 +/C1039G mice. Moreover, disruption in the elastic lamina was noted in the absence of aneurysms in pressurized aortas but not unpressurized aortas of Fbn1 +/C1039G mice. At the molecular level, this altered tissue behavior in non-aneurysmal descending aortas of Fbn1 +/C1039G mice was accompanied by an increasing trend of canonical but not noncanonical, transforming growth factor-β (TGFβ) signaling. Finally, assays of in vitro collagen lattice formation in mouse wild-type and TGFβ1-deficient embryonic fibroblasts indicate that TGFβ1 can regulate collagen organization. The ability to reveal the presence of altered biomechanics and microstructure coupled with subtle changes in TGFβ signaling provides a novel surrogate measure of tissue susceptibility to aneurysm before disease onset.
    Keywords aneurysm ; animal models ; aorta ; biomechanics ; collagen ; disease models ; disease resistance ; fibroblasts ; genes ; image analysis ; in vitro studies ; mice ; microstructure ; mutation ; patients ; transforming growth factor beta
    Language English
    Dates of publication 2012-01
    Size p. 267-277.
    Publishing place Springer-Verlag
    Document type Article
    ZDB-ID 125067-x
    ISSN 1432-0878 ; 0302-766X
    ISSN (online) 1432-0878
    ISSN 0302-766X
    DOI 10.1007/s00441-011-1270-y
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  4. Article ; Online: Altered tissue behavior of a non-aneurysmal descending thoracic aorta in the mouse model of Marfan syndrome.

    Haskett, Darren / Doyle, Jefferson J / Gard, Connie / Chen, Hwudaurw / Ball, Corbie / Estabrook, Mark A / Encinas, Alejandra C / Dietz, Harry C / Utzinger, Urs / Vande Geest, Jonathan P / Azhar, Mohamad

    Cell and tissue research

    2012  Volume 347, Issue 1, Page(s) 267–277

    Abstract: Aortic aneurysm is predominantly found in the ascending aorta in patients with Marfan syndrome (MFS). However, descending aortic disease has emerged as a problem since people are living longer because of improved medical and surgical management of the ... ...

    Abstract Aortic aneurysm is predominantly found in the ascending aorta in patients with Marfan syndrome (MFS). However, descending aortic disease has emerged as a problem since people are living longer because of improved medical and surgical management of the ascending aorta. Diagnostic procedures before disease onset and the mechanisms involved in the transition of normal aortic tissue to aneurysm remain unclear. We determined signs of descending aortic disease before disease onset in mice with a mutation in the fibrillin 1 gene (Fbn1(+/C1039G)), a validated mouse model of disease susceptibility and progression of aortic aneurysm of MFS. We analyzed a tubular unfixed non-aneurysmal descending thoracic aorta from 8-month-old wild-type and Fbn1(+/C1039G) mice by a tubular biaxial tester that works in conjunction with a two-photon nonlinear microscope. Fbn1(+/C1039G) mouse aorta was more compliant in the circumferential direction. Two-photon imaging showed defective organization of adventitial collagen fibers in the pressurized aortas of Fbn1(+/C1039G) mice. Moreover, disruption in the elastic lamina was noted in the absence of aneurysms in pressurized aortas but not unpressurized aortas of Fbn1(+/C1039G) mice. At the molecular level, this altered tissue behavior in non-aneurysmal descending aortas of Fbn1(+/C1039G) mice was accompanied by an increasing trend of canonical but not noncanonical, transforming growth factor-β (TGFβ) signaling. Finally, assays of in vitro collagen lattice formation in mouse wild-type and TGFβ1-deficient embryonic fibroblasts indicate that TGFβ1 can regulate collagen organization. The ability to reveal the presence of altered biomechanics and microstructure coupled with subtle changes in TGFβ signaling provides a novel surrogate measure of tissue susceptibility to aneurysm before disease onset.
    MeSH term(s) Animals ; Aorta, Thoracic/pathology ; Aorta, Thoracic/physiology ; Aortic Aneurysm, Thoracic/etiology ; Disease Models, Animal ; Fibrillin-1 ; Fibrillins ; Humans ; Marfan Syndrome/complications ; Marfan Syndrome/pathology ; Marfan Syndrome/physiopathology ; Mice ; Microfilament Proteins/genetics ; Microfilament Proteins/metabolism ; Stress, Mechanical ; Vascular Diseases/etiology ; Vascular Diseases/pathology
    Chemical Substances FBN1 protein, human ; Fbn1 protein, mouse ; Fibrillin-1 ; Fibrillins ; Microfilament Proteins
    Language English
    Publishing date 2012-01
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 125067-x
    ISSN 1432-0878 ; 0302-766X
    ISSN (online) 1432-0878
    ISSN 0302-766X
    DOI 10.1007/s00441-011-1270-y
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

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  5. Article ; Online: Generation of mice with a conditional allele for the transforming growth factor beta3 gene.

    Doetschman, Thomas / Georgieva, Teodora / Li, Hongqi / Reed, Thomas D / Grisham, Christina / Friel, Jacqueline / Estabrook, Mark A / Gard, Connie / Sanford, L P / Azhar, Mohamad

    Genesis (New York, N.Y. : 2000)

    2012  Volume 50, Issue 1, Page(s) 59–66

    Abstract: The transforming growth factor beta (TGFβ) pathway is involved in embryonic development and several inherited and acquired human diseases. The gene for TGFβ3 (Tgfb3) encodes one of the three ligands for TGFβ receptors. It is widely expressed in the ... ...

    Abstract The transforming growth factor beta (TGFβ) pathway is involved in embryonic development and several inherited and acquired human diseases. The gene for TGFβ3 (Tgfb3) encodes one of the three ligands for TGFβ receptors. It is widely expressed in the embryo and its mutation or misexpression is found in human diseases. Tgfb3-/- mice die at birth from cleft palate, precluding functional studies in adults. Here, we generated mice in which exon 6 of Tgfb3 was flanked with LoxP sites (Tgfb3flox/flox). The adult mice were normal and fertile. EIIa-Cre-mediated deletion of exon 6 in Tgfb3flox/flox mice efficiently generated Tgfb3 conditional knockout (Tgfb3cko/cko) mice which died at birth from the same cleft palate defect as Tgfb3-/- mice, indicating that the conditional and knockout alleles are functionally equivalent. This Tgfb3cko allele will now enable studies of TGFβ3 function in different cell or tissue types in embryonic development and during adulthood.
    MeSH term(s) Alleles ; Animals ; Cleft Palate/embryology ; Exons ; Female ; Gene Expression Regulation, Developmental ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Phenotype ; Real-Time Polymerase Chain Reaction ; Receptors, Transforming Growth Factor beta/metabolism ; Sequence Analysis, DNA ; Transforming Growth Factor beta3/genetics ; Transforming Growth Factor beta3/metabolism
    Chemical Substances Receptors, Transforming Growth Factor beta ; Transforming Growth Factor beta3
    Language English
    Publishing date 2012-01-06
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Technical Report
    ZDB-ID 2004544-X
    ISSN 1526-968X ; 1526-954X
    ISSN (online) 1526-968X
    ISSN 1526-954X
    DOI 10.1002/dvg.20789
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    In stock of ZB MED Cologne/Königswinter

    Zs.A 2772: Show issues Location:
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  6. Article ; Online: Transforming growth factor Beta2 is required for valve remodeling during heart development.

    Azhar, Mohamad / Brown, Kristen / Gard, Connie / Chen, Hwudaurw / Rajan, Sudarsan / Elliott, David A / Stevens, Mark V / Camenisch, Todd D / Conway, Simon J / Doetschman, Thomas

    Developmental dynamics : an official publication of the American Association of Anatomists

    2011  Volume 240, Issue 9, Page(s) 2127–2141

    Abstract: Although the function of transforming growth factor beta2 (TGFβ2) in epithelial mesenchymal transition (EMT) is well studied, its role in valve remodeling remains to be fully explored. Here, we used histological, morphometric, immunohistochemical and ... ...

    Abstract Although the function of transforming growth factor beta2 (TGFβ2) in epithelial mesenchymal transition (EMT) is well studied, its role in valve remodeling remains to be fully explored. Here, we used histological, morphometric, immunohistochemical and molecular approaches and showed that significant dysregulation of major extracellular matrix (ECM) components contributed to valve remodeling defects in Tgfb2(-/-) embryos. The data indicated that cushion mesenchymal cell differentiation was impaired in Tgfb2(-/-) embryos. Hyaluronan and cartilage link protein-1 (CRTL1) were increased in hyperplastic valves of Tgfb2(-/-) embryos, indicating increased expansion and diversification of cushion mesenchyme into the cartilage cell lineage during heart development. Finally, Western blot and immunohistochemistry analyses indicate that the activation of SMAD2/3 was decreased in Tgfb2(-/-) embryos during valve remodeling. Collectively, the data indicate that TGFβ2 promotes valve remodeling and differentiation by inducing matrix organization and suppressing cushion mesenchyme differentiation into cartilage cell lineage during heart development.
    MeSH term(s) Animals ; Cell Differentiation/genetics ; Cell Differentiation/physiology ; Extracellular Matrix/metabolism ; Heart/embryology ; Heart Valves/embryology ; Heart Valves/metabolism ; Immunohistochemistry ; Mesoderm/cytology ; Mice ; Mice, Knockout ; Real-Time Polymerase Chain Reaction ; Smad2 Protein/metabolism ; Smad3 Protein/metabolism ; Transforming Growth Factor beta2/genetics ; Transforming Growth Factor beta2/metabolism
    Chemical Substances Smad2 Protein ; Smad2 protein, mouse ; Smad3 Protein ; Transforming Growth Factor beta2
    Language English
    Publishing date 2011-07-20
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 1102541-4
    ISSN 1097-0177 ; 1058-8388
    ISSN (online) 1097-0177
    ISSN 1058-8388
    DOI 10.1002/dvdy.22702
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

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    Zs.MG 64: Show issues

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  7. Article: Gene targeted ablation of high molecular weight fibroblast growth factor-2.

    Azhar, Mohamad / Yin, Moying / Zhou, Ming / Li, Hongqi / Mustafa, Marwan / Nusayr, Eyad / Keenan, Jack B / Chen, Hwudaurw / Pawlosky, Sharon / Gard, Connie / Grisham, Christina / Sanford, L Philip / Doetschman, Tom

    Developmental dynamics : an official publication of the American Association of Anatomists

    2008  Volume 238, Issue 2, Page(s) 351–357

    Abstract: Fibroblast growth factor-2 (FGF2) is produced as high molecular weight isoforms (HMW) and a low molecular weight isoform (LMW) by means of alternative usage of translation start sites in a single Fgf2 mRNA. Although the physiological function of FGF2 and ...

    Abstract Fibroblast growth factor-2 (FGF2) is produced as high molecular weight isoforms (HMW) and a low molecular weight isoform (LMW) by means of alternative usage of translation start sites in a single Fgf2 mRNA. Although the physiological function of FGF2 and FGF2 LMW has been investigated in myocardial capillarogenesis during normal cardiac growth, the role of FGF2 HMW has not been determined. Here, we report the generation of FGF2 HMW-deficient mice in which FGF2 HMW isoforms are ablated by the Tag-and-Exchange gene targeting technique. These mice are normal and fertile with normal fecundity, and have a normal life span. Histological, immunohistochemical, and morphometric analyses indicate normal myocardial architecture, blood vessel, and cardiac capillary density in young adult FGF2 HMW-deficient mice. These mice along with the FGF2- and FGF2 LMW-deficient mice that we have generated previously will be very useful for elucidating the differential functions of FGF2 isoforms in pathophysiology of cardiovascular diseases.
    MeSH term(s) Animals ; Capillaries/physiology ; Coronary Vessels/metabolism ; Fibroblast Growth Factor 2/genetics ; Fibroblast Growth Factor 2/physiology ; Mice ; Mice, Knockout ; Myocardium/metabolism ; Protein Isoforms/genetics ; Protein Isoforms/physiology
    Chemical Substances Protein Isoforms ; Fibroblast Growth Factor 2 (103107-01-3)
    Language English
    Publishing date 2008-12-22
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 1102541-4
    ISSN 1097-0177 ; 1058-8388
    ISSN (online) 1097-0177
    ISSN 1058-8388
    DOI 10.1002/dvdy.21835
    Database MEDical Literature Analysis and Retrieval System OnLINE

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