Generation of conditional Mef2cloxP/loxP mice for temporal‐ and tissue‐specific analyses

Mef2c belongs to the myocyte enhancer factor 2 (MEF2) family of MADS‐box containing transcription factors, which have been shown to be important for various processes involved in cell differentiation, cell survival, and apoptosis. Previous gene‐targeting studies have demonstrated a role for mef2c in early heart development since mice lacking mef2c die at embryonic day 9.5 due to cardiac and vascular defects. Since the early embryonic lethality of mef2c prevents an examination of its role in the later stages of heart development, conditional mef2cloxP/loxP mice were generated to allow for temporal‐ and tissue‐specific analyses. We report here that general Cre recombinase‐mediated removal of the second coding exon of mef2c phenocopied the original mef2c null. Additionally, myocardial‐specific removal of mef2c resulted in viable offspring, demonstrating that while mef2c is required for the early development of the heart, it is not necessary for the formation of the heart after looping morphogenesis. genesis 43:43–48, 2005. © 2005 Wiley‐Liss, Inc.

[1]  B. Calnan,et al.  Regulation of the Nur77 orphan steroid receptor in activation-induced apoptosis , 1995, Molecular and cellular biology.

[2]  G. Lyons,et al.  Mef2 gene expression marks the cardiac and skeletal muscle lineages during mouse embryogenesis. , 1994, Development.

[3]  B. Black,et al.  Transcriptional control of muscle development by myocyte enhancer factor-2 (MEF2) proteins. , 1998, Annual review of cell and developmental biology.

[4]  Philippe Soriano Generalized lacZ expression with the ROSA26 Cre reporter strain , 1999, Nature Genetics.

[5]  Jun O. Liu,et al.  Integration of calcineurin and MEF2 signals by the coactivator p300 during T‐cell apoptosis , 2000, The EMBO journal.

[6]  G. Lyons,et al.  Expression of mef2 genes in the mouse central nervous system suggests a role in neuronal maturation , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[7]  R. Behringer,et al.  Endocardial cushion and myocardial defects after cardiac myocyte-specific conditional deletion of the bone morphogenetic protein receptor ALK3 , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[8]  B. Hogan,et al.  Manipulating the mouse embryo: A laboratory manual , 1986 .

[9]  K. Heidenreich,et al.  Myocyte enhancer factor-2 transcription factors in neuronal differentiation and survival , 2004, Molecular Neurobiology.

[10]  C. Bucana,et al.  Control of mouse cardiac morphogenesis and myogenesis by transcription factor MEF2C. , 1997, Science.

[11]  A. Moorman,et al.  Chamber formation and morphogenesis in the developing mammalian heart. , 2000, Developmental biology.

[12]  J. Pessin,et al.  The MEF2A Isoform Is Required for Striated Muscle-specific Expression of the Insulin-responsive GLUT4 Glucose Transporter* , 2000, The Journal of Biological Chemistry.

[13]  C. Drake,et al.  The transcription factor MEF2C-null mouse exhibits complex vascular malformations and reduced cardiac expression of angiopoietin 1 and VEGF. , 1999, Developmental biology.

[14]  E. Olson,et al.  Myocyte enhancer binding factor-2 expression and activity in vascular smooth muscle cells. Association with the activated phenotype. , 1996, Circulation research.

[15]  T. Chatila,et al.  Ca2+-dependent Gene Expression Mediated by MEF2 Transcription Factors* , 2000, The Journal of Biological Chemistry.

[16]  Philippe Soriano,et al.  Widespread recombinase expression using FLPeR (Flipper) mice , 2000, Genesis.

[17]  H. Youn,et al.  Apoptosis of T cells mediated by Ca2+-induced release of the transcription factor MEF2. , 1999, Science.

[18]  D. Srivastava,et al.  The Hand1 and Hand2 transcription factors regulate expansion of the embryonic cardiac ventricles in a gene dosage-dependent manner , 2004, Development.

[19]  G. Fishman,et al.  Regulation of peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) and mitochondrial function by MEF2 and HDAC5 , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[20]  B. A. French,et al.  Gene recombination in postmitotic cells. Targeted expression of Cre recombinase provokes cardiac-restricted, site-specific rearrangement in adult ventricular muscle in vivo. , 1997, The Journal of clinical investigation.

[21]  K. Chien,et al.  Ventricular muscle-restricted targeting of the RXRalpha gene reveals a non-cell-autonomous requirement in cardiac chamber morphogenesis. , 1998, Development.

[22]  R. Kothary,et al.  MEF2 is upregulated during cardiac hypertrophy and is required for normal post-natal growth of the myocardium , 1999, Current Biology.

[23]  B. Black,et al.  Mitochondrial deficiency and cardiac sudden death in mice lacking the MEF2A transcription factor , 2002, Nature Medicine.

[24]  S. Subramanian,et al.  Early expression of the different isoforms of the myocyte enhancer factor-2 (MEF2) protein in myogenic as well as non-myogenic cell lineages during mouse embryogenesis , 1996, Mechanisms of Development.

[25]  F. Alt,et al.  Efficient in vivo manipulation of mouse genomic sequences at the zygote stage. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[26]  M E Greenberg,et al.  Neuronal activity-dependent cell survival mediated by transcription factor MEF2. , 1999, Science.

[27]  A. Sharrocks,et al.  The MADS-box family of transcription factors. , 1995, European journal of biochemistry.

[28]  G. Lyons,et al.  Requirement of the MADS-box transcription factor MEF2C for vascular development. , 1998, Development.

[29]  S. Guruswamy,et al.  Myocyte enhancer factor 2 (MEF2)-binding site is required for GLUT4 gene expression in transgenic mice. Regulation of MEF2 DNA binding activity in insulin-deficient diabetes. , 1998, The Journal of biological chemistry.