Targeted Disruption of the Mouse Estrogen Sulfotransferase Gene Reveals a Role of Estrogen Metabolism in Intracrine and Paracrine Estrogen Regulation

The leukotrienes (LTs) are metabolic products of arachidonic acid via the 5-lipoxygenase (5-LO) pathway. The biological activities of LTs suggest that they are mediators of acute inflammatory and immediate hypersensitivity responses. In particular, the 5-LO activation has been proposed to be an important regulator for pathogenesis in multicellular organisms. The role of LTs in tissue damage, associated with septic and nonseptic shock and ischemia-reperfusion, has been extensively studied by the use of 5-LO inhibitors, receptor antagonists, and mice with a targeted disruption of the 5-LO gene (5-LOKO). In particular, several data indicate that LTs regulate neutrophil trafficking in damaged tissue in shock and ischemia-reperfusion, mainly through the modulation of adhesion molecule expression. This concept may provide new insights into the interpretation of the protective effect of 5-LO inhibition, which may be useful in the therapy of pathological conditions associated with septic and nonseptic shock and ischemia-reperfusion injury.

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Akt2 Regulates Cardiac Metabolism and Cardiomyocyte Survival

Journal of Biological Chemistry ◽

10.1074/jbc.m513087200 ◽

2006 ◽

Vol 281 (43) ◽

pp. 32841-32851 ◽

Cited By ~ 110

Author(s):

Brian DeBosch ◽

Nandakumar Sambandam ◽

Carla Weinheimer ◽

Michael Courtois ◽

Anthony J. Muslin

Keyword(s):

Pressure Overload ◽

Cardiac Metabolism ◽

Cellular Protein ◽

Growth Responses ◽

Infarct Zone ◽

Targeted Disruption ◽

Cellular Processes ◽

Ligand Stimulation ◽

And Function

The Akt family of serine-threonine kinases participates in diverse cellular processes, including the promotion of cell survival, glucose metabolism, and cellular protein synthesis. All three known Akt family members, Akt1, Akt2 and Akt3, are expressed in the myocardium, although Akt1 and Akt2 are most abundant. Previous studies demonstrated that Akt1 and Akt3 overexpression results in enhanced myocardial size and function. Yet, little is known about the role of Akt2 in modulating cardiac metabolism, survival, and growth. Here, we utilize murine models with targeted disruption of the akt2 or the akt1 genes to demonstrate that Akt2, but not Akt1, is required for insulin-stimulated 2-[3H]deoxyglucose uptake and metabolism. In contrast, akt2-/- mice displayed normal cardiac growth responses to provocative stimulation, including ligand stimulation of cultured cardiomyocytes, pressure overload by transverse aortic constriction, and myocardial infarction. However, akt2-/- mice were found to be sensitized to cardiomyocyte apoptosis in response to ischemic injury, and apoptosis was significantly increased in the peri-infarct zone of akt2-/- hearts 7 days after occlusion of the left coronary artery. These results implicate Akt2 in the regulation of cardiomyocyte metabolism and survival.

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Regulation of Estrogen Sulfotransferase Expression by Confluence of MCF10A Breast Epithelial Cells: Role of the Aryl Hydrocarbon Receptor

Journal of Pharmacology and Experimental Therapeutics ◽

10.1124/jpet.111.185173 ◽

2011 ◽

Vol 339 (2) ◽

pp. 597-606 ◽

Cited By ~ 9

Author(s):

Jiaqi Fu ◽

Hailin Fang ◽

Michelle Paulsen ◽

Mats Ljungman ◽

Thomas A. Kocarek ◽

...

Keyword(s):

Epithelial Cells ◽

Aryl Hydrocarbon Receptor ◽

Estrogen Sulfotransferase ◽

Breast Epithelial Cells ◽

Aryl Hydrocarbon ◽

Breast Epithelial

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Generation of a Mouse Model for Arginase II Deficiency by Targeted Disruption of the Arginase II Gene

Molecular and Cellular Biology ◽

10.1128/mcb.21.3.811-813.2001 ◽

2001 ◽

Vol 21 (3) ◽

pp. 811-813 ◽

Cited By ~ 93

Author(s):

Ou Shi ◽

Sidney M. Morris ◽

Huda Zoghbi ◽

Carl W. Porter ◽

William E. O'Brien

Keyword(s):

Urea Cycle ◽

Elevated Plasma ◽

Wild Type ◽

Targeted Disruption ◽

Arginase Ii ◽

Plasma Arginine ◽

Physiologic Role ◽

Deficient Mice ◽

Arginase I

ABSTRACT Mammals express two isoforms of arginase, designated types I and II. Arginase I is a component of the urea cycle, and inherited defects in arginase I have deleterious consequences in humans. In contrast, the physiologic role of arginase II has not been defined, and no deficiencies in arginase II have been identified in humans. Mice with a disruption in the arginase II gene were created to investigate the role of this enzyme. Homozygous arginase II-deficient mice were viable and apparently indistinguishable from wild-type mice, except for an elevated plasma arginine level which indicates that arginase II plays an important role in arginine homeostasis.

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MicroRNA as Crucial Regulators of Gene Expression in Estradiol-Treated Human Endothelial Cells

Cellular Physiology and Biochemistry ◽

10.1159/000487910 ◽

2018 ◽

Vol 45 (5) ◽

pp. 1878-1892 ◽

Cited By ~ 23

Author(s):

Xavier Vidal-Gómez ◽

Daniel Pérez-Cremades ◽

Ana Mompeón ◽

Ana Paula Dantas ◽

Susana Novella ◽

...

Keyword(s):

Endothelial Cells ◽

Endothelial Function ◽

Pathway Analysis ◽

Mirna Expression ◽

Umbilical Vein ◽

Mirna Gene ◽

Mirna Profile ◽

Human Endothelial Cells ◽

Estrogen Regulation

Background/Aims: Estrogen signalling plays an important role in vascular biology as it modulates vasoactive and metabolic pathways in endothelial cells. Growing evidence has also established microRNA (miRNA) as key regulators of endothelial function. Nonetheless, the role of estrogen regulation on miRNA profile in endothelial cells is poorly understood. In this study, we aimed to determine how estrogen modulates miRNA profile in human endothelial cells and to explore the role of the different estrogen receptors (ERα, ERβ and GPER) in the regulation of miRNA expression by estrogen. Methods: We used miRNA microarrays to determine global miRNA expression in human umbilical vein endothelial cells (HUVEC) exposed to a physiological concentration of estradiol (E2; 1 nmol/L) for 24 hours. miRNA-gene interactions were computationally predicted using Ingenuity Pathway Analysis and changes in miRNA levels were validated by qRT-PCR. Role of ER in the E2-induced miRNA was additionally confirmed by using specific ER agonists and antagonists. Results: miRNA array revealed that expression of 114 miRNA were significantly modified after E2 exposition. Further biological pathway analysis revealed cell death and survival, lipid metabolism, reproductive system function, as the top functions regulated by E2. We validated changes in the most significantly increased (miR-30b-5p, miR-487a-5p, miR-4710, miR-501-3p) and decreased (miR-378h and miR-1244) miRNA and the role of ER in these E2-induced miRNA was determined. Results showed that both classical, ERα and ERβ, and membrane-bound ER, GPER, differentially regulated specific miRNA. In silico analysis of validated miRNA promoters identified specific ER binding sites. Conclusion: Our findings identify differentially expressed miRNA pathways linked to E2 in human endothelial cells through ER, and provide new insights by which estrogen can modulate endothelial function.

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Targeted disruption of the murine junD gene results in multiple defects in male reproductive function

Development ◽

10.1242/dev.127.1.143 ◽

2000 ◽

Vol 127 (1) ◽

pp. 143-153 ◽

Cited By ~ 2

Author(s):

D. Thepot ◽

J.B. Weitzman ◽

J. Barra ◽

D. Segretain ◽

M.G. Stinnakre ◽

...

Keyword(s):

Reproductive Function ◽

Postnatal Growth ◽

Mrna Levels ◽

Targeted Disruption ◽

Male Reproductive Function ◽

Developing Heart ◽

Redundant Functions ◽

Transcription Factor Complex

JunD is one of three mammalian Jun proteins that contribute to the AP-1 transcription factor complex. Distinct regulation and functions have been proposed for each Jun member, but less is known about the biological functions of each of these proteins in vivo. To investigate the role of JunD, we have inactivated the murine gene by replacement with a bacterial lacZ reporter gene. Embryonic JunD expression was initially detected in the developing heart and cardiovascular system. Subsequent broadening phases of JunD expression were observed during embryonic development and expression in the adult was widespread in many tissues and cell lineages. Mutant animals lack JunD mRNA and protein and showed no evidence of upregulation of c-Jun and JunB mRNA levels. In contrast to the other two Jun members, homozygous JunD−/− mutant animals were viable and appeared healthy. However, homozygous JunD−/− animals showed a reduced postnatal growth. Furthermore, JunD−/− males exhibited multiple age-dependent defects in reproduction, hormone imbalance and impaired spermatogenesis with abnormalities in head and flagellum sperm structures. No defects in fertility were observed in JunD−/− female animals. These results provide evidence for redundant functions for members of the Jun family during development and specific functions for JunD in male reproductive function.

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Mice homozygous for a targeted disruption of Hoxd-3 (Hox-4.1) exhibit anterior transformations of the first and second cervical vertebrae, the atlas and the axis

Development ◽

10.1242/dev.119.3.579 ◽

1993 ◽

Vol 119 (3) ◽

pp. 579-595 ◽

Cited By ~ 8

Author(s):

B.G. Condie ◽

M.R. Capecchi

Keyword(s):

Hox Genes ◽

Es Cells ◽

Cervical Vertebrae ◽

Expression Patterns ◽

Cervical Vertebra ◽

Targeted Disruption ◽

Paralogous Genes ◽

Variable Extent ◽

Basioccipital Bone

Gene targeting in embryo-derived stem (ES) cells was used to generate mice with a disruption in the homeobox-containing gene Hoxd-3 (Hox-4.1). Mice homozygous for this mutation show a radically remodeled craniocervical joint. The anterior arch of the atlas is transformed to an extension of the basioccipital bone of the skull. The lateral masses of the atlas also assume a morphology more closely resembling the exoccipitals and, to a variable extent, fuse with the exoccipitals. Formation of the second cervical vertebra, the axis, is also affected. The dens and the superior facets are deleted, and the axis shows ‘atlas-like’ characteristics. An unexpected observation is that different parts of the same vertebra are differentially affected by the loss of Hoxd-3 function. Some parts are deleted, others are homeotically transformed to more anterior structures. These observations suggest that one role of Hox genes may be to differentially control the proliferation rates of the mesenchymal condensations that give rise to the vertebral cartilages. Within the mouse Hox complex, paralogous genes not only encode very similar proteins but also often exhibit very similar expression patterns. Therefore, it has been postulated that paralogous Hox genes would perform similar roles. Surprisingly, however, no tissues or structures are affected in common by mutations in the two paralogous genes, Hoxa-3 and Hoxd-3.

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Impaired FcϵRI stability, signaling, and effector functions in murine mast cells lacking glycosylphosphatidylinositol-anchored proteins

Blood ◽

10.1182/blood-2011-02-338053 ◽

2011 ◽

Vol 118 (16) ◽

pp. 4377-4383 ◽

Cited By ~ 6

Author(s):

Wouter L. W. Hazenbos ◽

Ping Wu ◽

Jeffrey Eastham-Anderson ◽

Taroh Kinoshita ◽

Eric J. Brown

Keyword(s):

Mast Cells ◽

Targeted Disruption ◽

Potential Therapeutic Target ◽

Effector Functions ◽

Ige Mediated ◽

Interchain Interactions ◽

Β Chain ◽

Stimulation Of

Abstract A key event and potential therapeutic target in allergic and asthmatic diseases is signaling by the IgE receptor FcϵRI, which depends on its interactions with Src family kinases (SFK). Here we tested the hypothesis that glycosylphosphatidylinositiol-anchored proteins (GPI-AP) are involved in FcϵRI signaling, based on previous observations that GPI-AP colocalize with and mediate activation of SFK. We generated mice with a hematopoietic cell-specific GPI-AP deficiency by targeted disruption of the GPI biosynthesis gene PigA. In these mice, IgE-mediated passive cutaneous anaphylaxis was largely abolished. PigA-deficient mast cells cultured from these mice showed impaired degranulation in response to stimulation with IgE and antigen in vitro, despite normal IgE binding and antigen-induced FcϵRI aggregation. On stimulation of these cells with IgE and antigen, coprecipitation of the FcϵRI α-chain with the γ-chain and β-chain was markedly reduced. As a result, IgE/antigen–induced FcϵRI-Lyn association and γ-chain tyrosine phosphorylation were both impaired in PigA-deficient cells. These data provide genetic evidence for an unanticipated key role of GPI-AP in FcϵRI interchain interactions and early FcϵRI signaling events, necessary for antigen-induced mast cell degranulation.

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The Mouse Polyubiquitin Gene Ubb Is Essential for Meiotic Progression

Molecular and Cellular Biology ◽

10.1128/mcb.01566-07 ◽

2007 ◽

Vol 28 (3) ◽

pp. 1136-1146 ◽

Cited By ~ 61

Author(s):

Kwon-Yul Ryu ◽

Shamim A. Sinnar ◽

Laura G. Reinholdt ◽

Sergio Vaccari ◽

Susan Hall ◽

...

Keyword(s):

Germ Cells ◽

Striking Similarity ◽

Eukaryotic Evolution ◽

Targeted Disruption ◽

Male And Female ◽

Meiotic Progression ◽

Polyubiquitin Gene ◽

Polyubiquitin Genes ◽

Meiosis I

ABSTRACT Ubiquitin is encoded in mice by two polyubiquitin genes, Ubb and Ubc, that are considered to be stress inducible and two constitutively expressed monoubiquitin (Uba) genes. Here we report that targeted disruption of Ubb results in male and female infertility due to failure of germ cells to progress through meiosis I and hypogonadism. In the absence of Ubb, spermatocytes and oocytes arrest during meiotic prophase, before metaphase of the first meiotic division. Although cellular ubiquitin levels are believed to be maintained by a combination of functional redundancy among the four ubiquitin genes, stress inducibility of the two polyubiquitin genes, and ubiquitin recycling by proteasome-associated isopeptidases, our results indicate that ubiquitin is required for and consumed during meiotic progression. The striking similarity of the meiotic phenotype in Ubb −/− germ cells to the sporulation defect in fission yeast (Schizosaccharomyces pombe) lacking a polyubiquitin gene suggests that a meiotic role of the polyubiquitin gene has been conserved throughout eukaryotic evolution.

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