scholarly journals Loss of mRor1 Enhances the Heart and Skeletal Abnormalities in mRor2-Deficient Mice: Redundant and Pleiotropic Functions of mRor1 and mRor2 Receptor Tyrosine Kinases

2001 ◽  
Vol 21 (24) ◽  
pp. 8329-8335 ◽  
Author(s):  
Masashi Nomi ◽  
Isao Oishi ◽  
Shuichi Kani ◽  
Hiroaki Suzuki ◽  
Takeru Matsuda ◽  
...  
Keyword(s):  
Tyrosine Kinases ◽  
Double Mutant ◽  
Receptor Tyrosine Kinases ◽  
Cardiac Development ◽  
Mutant Mice ◽  
Pubic Bone ◽  
Respiratory Dysfunction ◽  
Skeletal Abnormalities ◽  
Pleiotropic Functions ◽  
Deficient Mice

ABSTRACT The mammalian Ror family of receptor tyrosine kinases consists of two structurally related proteins, Ror1 and Ror2. We have shown that mRor2-deficient mice exhibit widespread skeletal abnormalities, ventricular septal defects in the heart, and respiratory dysfunction, leading to neonatal lethality (S. Takeuchi, K. Takeda, I. Oishi, M. Nomi, M. Ikeya, K. Itoh, S. Tamura, T. Ueda, T. Hatta, H. Otani, T. Terashima, S. Takada, H. Yamamura, S. Akira, and Y. Minami, Genes Cells 5:71–78, 2000). Here we show thatmRor1-deficient mice have no apparent skeletal or cardiac abnormalities, yet they also die soon after birth due to respiratory dysfunction. Interestingly,mRor1/mRor2 double mutant mice show markedly enhanced skeletal abnormalities compared withmRor2 mutant mice. Furthermore, double mutant mice also exhibit defects not observed in mRor2 mutant mice, including a sternal defect, dysplasia of the symphysis of the pubic bone, and complete transposition of the great arteries. These results indicate that mRor1 and mRor2 interact genetically in skeletal and cardiac development.

scholarly journals Nkx2.5 and Nkx2.6, Homologs ofDrosophila tinman, Are Required for Development of the Pharynx

2001 ◽  
Vol 21 (13) ◽  
pp. 4391-4398 ◽  
Author(s):  
Makoto Tanaka ◽  
Martina Schinke ◽  
Hai-Sun Liao ◽  
Naohito Yamasaki ◽  
Seigo Izumo
Keyword(s):  
Double Mutant ◽  
Cardiac Development ◽  
Critical Role ◽  
Homeobox Genes ◽  
Mutant Mice ◽  
Ventral Region ◽  
Early Stages ◽  
Endodermal Cells

ABSTRACT Nkx2.5 and Nkx2.6 are murine homologs of Drosophilatinman. Their genes are expressed in the ventral region of the pharynx at early stages of embryogenesis. However, no abnormalities in the pharynges of embryos with mutations in either Nkx2.5 or Nkx2.6 have been reported. To examine the function of Nkx2.5 and Nkx2.6 in the formation of the pharynx, we generated and analyzed Nkx2.5 and Nkx2.6 double-mutant mice. Interestingly, in the double-mutant embryos, the pharynx did not form properly. Pharyngeal endodermal cells were largely missing, and the mutant pharynx was markedly dilated. Moreover, we observed enhanced apoptosis and reduced proliferation in pharyngeal endodermal cells of the double-mutant embryos. These results demonstrated a critical role of the NK-2 homeobox genes in the differentiation, proliferation, and survival of pharyngeal endodermal cells. Furthermore, the development of the atrium was less advanced in the double-mutant embryos, indicating that these two genes are essential for both pharyngeal and cardiac development.

scholarly journals Genetic Evidence for Functional Dependency of p18Ink4c on Cdk4

2004 ◽  
Vol 24 (15) ◽  
pp. 6653-6664 ◽  
Author(s):  
Xin-Hai Pei ◽  
Feng Bai ◽  
Tateki Tsutsui ◽  
Hiroaki Kiyokawa ◽  
Yue Xiong
Keyword(s):  
Double Mutant ◽  
Wide Spectrum ◽  
Genetic Evidence ◽  
Mutant Mice ◽  
Cdk Inhibitors ◽  
Cyclin D ◽  
Cyclin Dependent Kinase ◽  
Single Mutant ◽  
Cell Growth And Proliferation ◽  
Deficient Mice

ABSTRACT The INK4 family of cyclin-dependent kinase (CDK) inhibitors negatively regulates cyclin D-dependent CDK4 and CDK6 and induces the growth-suppressive function of Rb family proteins. Mutations in the Cdk4 gene conferring INK4 resistance are associated with familial and sporadic melanoma in humans and result in a wide spectrum of tumors in mice, suggesting that INK4 is a major regulator of CDK4. Mice lacking the Cdk4 gene exhibit various defects in many organs associated with hypocellularity, whereas loss of the p18 Ink4c gene results in widespread hyperplasia and organomegaly. To genetically test the notion that the function of INK4 is dependent on CDK4, we generated p18; Cdk4 double-mutant mice and examined the organs and tissues which developed abnormalities when either gene is deleted. We show here that, in all organs we have examined, including pituitary, testis, pancreas, kidney, and adrenal gland, hyperproliferative phenotypes associated with p18 loss were canceled. The double-mutant mice exhibited phenotypes very close to or indistinguishable from that of Cdk4 single-mutant mice. Mice lacking p27 Kip1 develop widespread hyperplasia and organomegaly similar to those developed by p18-deficient mice. The p27; Cdk4 double-mutant mice, however, displayed phenotypes intermediate between those of p27 and Cdk4 single-mutant mice. These results provide genetic evidence that in mice p18 Ink4c and p27 Kip1 mediate the transduction of different cell growth and proliferation signals to CDK4 and that p18 Ink4c is functionally dependent on CDK4.

P-selectin and ICAM-1 mediate endotoxin-induced neutrophil recruitment and injury to the lung and liver

1999 ◽  
Vol 277 (2) ◽  
pp. L310-L319 ◽  
Author(s):  
M. Kamochi ◽  
F. Kamochi ◽  
Y. B. Kim ◽  
S. Sawh ◽  
J. M. Sanders ◽  
...  
Keyword(s):  
Double Mutant ◽  
Leukocyte Adhesion ◽  
Lavage Fluid ◽  
Mutant Mice ◽  
Organ Injury ◽  
Intercellular Adhesion Molecule ◽  
Myeloperoxidase Activity ◽  
Electron Microscopic ◽  
Systemic Endotoxemia ◽  
Deficient Mice

The role of leukocyte adhesion molecules in endotoxin-induced organ injury was evaluated by administering intraperitoneal Salmonella enteritidislipopolysaccharide (LPS) to wild-type (WT) mice, P-selectin-deficient mice, intercellular adhesion molecule (ICAM)-1-deficient mice, and P-selectin-ICAM-1 double-mutant mice. In WT mice, there was a sevenfold increase in the number of neutrophils present in the pulmonary vascular lavage fluid, and there were sevenfold more intracapillary neutrophils by electron-microscopic (EM) morphometry at 4 h after intraperitoneal LPS compared with that in control mice. Extravascular albumin accumulation increased approximately twofold in the lungs and liver of WT mice treated with LPS. In the double-mutant mice, although overall mortality after intraperitoneal LPS was not attenuated, there was a significant delay in mortality in the P-selectin-ICAM-1-deficient mutants compared with that in WT mice after intraperitoneal LPS ( P < 0.01). Moreover, compared with LPS-treated WT mice, lung and liver extravascular albumin accumulation was significantly lower in LPS-treated P-selectin-ICAM-1 double-mutant mice. Lung myeloperoxidase activity, normalized per 1,000 circulating neutrophils, increased after endotoxin in WT and P-selectin-deficient mice but not in P-selectin-ICAM-1 double-mutant mice. In addition, lung and liver myeloperoxidase activity per 1,000 circulating neutrophils in endotoxin-treated ICAM-1-deficient mice and P-selectin-ICAM-1 double mutants was significantly lower compared with that in endotoxin-treated WT mice. These data suggest that P-selectin and ICAM-1 significantly contribute to lung and liver injury after systemic endotoxemia.

scholarly journals Direct Association between Mouse PERIOD and CKIε Is Critical for a Functioning Circadian Clock

2004 ◽  
Vol 24 (2) ◽  
pp. 584-594 ◽  
Author(s):  
Choogon Lee ◽  
David R. Weaver ◽  
Steven M. Reppert
Keyword(s):  
Circadian Clock ◽  
Double Mutant ◽  
Mutant Mice ◽  
Chimeric Proteins ◽  
Posttranslational Regulation ◽  
Dependent Manner ◽  
Clock Proteins ◽  
Deficient Mice

ABSTRACT The mPER1 and mPER2 proteins have important roles in the circadian clock mechanism, whereas mPER3 is expendable. Here we examine the posttranslational regulation of mPER3 in vivo in mouse liver and compare it to the other mPER proteins to define the salient features required for clock function. Like mPER1 and mPER2, mPER3 is phosphorylated, changes cellular location, and interacts with other clock proteins in a time-dependent manner. Consistent with behavioral data from mPer2/3 and mPer1/3 double-mutant mice, either mPER1 or mPER2 alone can sustain rhythmic posttranslational events. However, mPER3 is unable to sustain molecular rhythmicity in mPer1/2 double-mutant mice. Indeed, mPER3 is always cytoplasmic and is not phosphorylated in the livers of mPer1-deficient mice, suggesting that mPER3 is regulated by mPER1 at a posttranslational level. In vitro studies with chimeric proteins suggest that the inability of mPER3 to support circadian clock function results in part from lack of direct and stable interaction with casein kinase Iε (CKIε). We thus propose that the CKIε-binding domain is critical not only for mPER phosphorylation but also for a functioning circadian clock.

scholarly journals Relative Contributions of Innate and Acquired Host Responses to Bacterial Control and Arthritis Development in Lyme Disease

2005 ◽  
Vol 73 (1) ◽  
pp. 657-660 ◽  
Author(s):  
Xiaohui Wang ◽  
Ying Ma ◽  
John H. Weis ◽  
James F. Zachary ◽  
Carsten J. Kirschning ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Double Mutant ◽  
Mutant Mice ◽  
Host Responses ◽  
Relative Importance ◽  
Toll Like Receptor ◽  
Host Defenses ◽  
Arthritis Severity ◽  
Toll Like Receptor 2 ◽  
Deficient Mice

ABSTRACT TLR2−/−/scid double-mutant mice were infected with B. burgdorferi to assess the relative importance of acquired and innate host defenses. Although spirochete levels at 4 weeks were lower in TLR2−/− mice than in TLR2−/−/scid mice, the increased arthritis severity of TLR2 (Toll-like receptor 2)-deficient mice was reduced by the presence of the scid mutation.

scholarly journals T cells, but not B cells, are required for bowel inflammation in interleukin 2-deficient mice.

1995 ◽  
Vol 182 (5) ◽  
pp. 1567-1572 ◽  
Author(s):  
A Ma ◽  
M Datta ◽  
E Margosian ◽  
J Chen ◽  
I Horak
Keyword(s):  
T Cells ◽  
B Cells ◽  
Bowel Disease ◽  
Double Mutant ◽  
Intestinal Inflammation ◽  
Interleukin 2 ◽  
Mutant Mice ◽  
Immunodeficient Mice ◽  
Inflammatory Bowel ◽  
Deficient Mice

Interleukin-2 (IL-2)-deficient (IL-2-/-) mice develop anemia and colonic inflammatory bowel disease. To elucidate the mechanism of this disease, we have bred IL-2-/- mice to two strains of immunodeficient mice, RAG-2-deficient (RAG-2-/-, lacking B and T cells) and JH-deficient mice (JH-/-, lacking B cells). IL-2-/-, RAG-2-/- double-mutant mice are disease free, while IL-2-/-, JH-/- double-mutant mice succumb to bowel disease at the same rate as IL-2-/- littermates. IL-2-/-, JH-/- mice do not, however, succumb to anemia. Thus, spontaneous intestinal inflammation in IL-2-/- mice requires mature T cells, not B cells, while anemia is dependent on B cells.

The residual megakaryocyte and platelet production in c-Mpl–deficient mice is not dependent on the actions of interleukin-6, interleukin-11, or leukemia inhibitory factor

Blood ◽  
2000 ◽  
Vol 95 (2) ◽  
pp. 528-534 ◽  
Author(s):  
Timothy Gainsford ◽  
Harshal Nandurkar ◽  
Donald Metcalf ◽  
Lorraine Robb ◽  
C. Glenn Begley ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Steady State ◽  
Leukemia Inhibitory Factor ◽  
Double Mutant ◽  
Essential Role ◽  
Physiological Role ◽  
Mutant Mice ◽  
Platelet Production ◽  
Interleukin 11 ◽  
Deficient Mice

Mice lacking thrombopoietin (TPO) or its receptor c-Mpl are severely thrombocytopenic, consistent with a dominant physiological role for this cytokine in megakaryocytopoiesis. However, these mice remain healthy and show no signs of spontaneous hemorrhage, implying that TPO-independent mechanisms for platelet production exist and are sufficient for hemostasis. To investigate the roles of cytokines that act through the gp130 signaling chain in the residual platelet production of mpl-/- mice, mpl-/-IL-6-/-, mpl-/-LIF-/-, andmpl-/-IL-11R-/-double-mutant mice were generated. In each of these compound mutants, the number of circulating platelets was no lower than that observed in mice lacking only the c-mpl gene. Moreover, the deficits in the numbers of megakaryocytes and megakaryocyte progenitor cells in the bone marrow and spleen were no further exacerbated inmpl-/-IL-6-/-,mpl-/-LIF-/-, ormpl-/-IL-11R-/-double-mutant mice compared with those in Mpl-deficient animals. In single IL-6-/-, LIF-/-, andIL-11R-/- mutant mice, platelet production was normal. These data establish that, as single regulators, IL-6, IL-11, and LIF have no essential role in normal steady-state megakaryocytopoiesis, and are not required for the residual megakaryocyte and platelet production seen in thec-mpl-/- mouse.

scholarly journals TMIC-62. FYN, AN EFFECTOR OF ONCOGENIC RECEPTOR TYROSINE KINASES SIGNALING IN GLIOBLASTOMA, INHIBITS ANTI-GLIOMA IMMUNE RESPONSES: IMPLICATIONS FOR IMMUNOTHERAPY

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi261-vi261
Author(s):  
Andrea Comba ◽  
Patrick J Dunn ◽  
Anna E Argento ◽  
Padma Kadiyala ◽  
Maria Ventosa ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Immune Responses ◽  
Tumor Progression ◽  
Tyrosine Kinases ◽  
Receptor Tyrosine Kinases ◽  
Checkpoint Inhibitors ◽  
Sleeping Beauty ◽  
Genetically Engineered ◽  
Immune Functions ◽  
Deficient Mice

Abstract There is currently much excitement for the use of immunotherapies in cancer. In spite of positive results using checkpoint inhibitors in melanoma and CAR T cells in leukemias, these strategies have not yet achieved robust clinical responses in human gliomas. A powerful inhibitory microenvironment is thought to be the culprit. Mechanisms that determine the inhibitory microenvironment remain poorly understood. Herein we demonstrate that FYN, a downstream target of receptor tyrosine kinases signaling, inhibits the anti-glioma immune response. We utilized genetically engineered mouse models (GEMM) of glioma based on the Sleeping Beauty Transposon method. We examined the activities of FYN in NP (N-ras + shp53), NPA (NP + shATRX), and NPD (NP + PDGF overexpression) tumors. We also generated FYN knockdown glioma stem cells in vitro to induce gliomas in immune-competent and immune-deficient mice of varying genetic backgrounds (NSG, CD8-/-). Flow cytometry was used to characterize immune cells within the glioma microenvironment. Our results show that FYN knockdown in NP, NPA, or NPD GEMM of glioma reduced tumor progression and increased survival by 25–77%. GSEA analysis of differential expressed genes of WT vs. FYN knockdown gliomas revealed enrichments of gene ontologies related to immune functions. In NSG and CD8-/- immune-deficient mice, FYN knockdown failed to inhibits tumor growth and increase animal survival. These results suggest that FYN enhances tumor progression through changes in anti-glioma immune activation. Examination of tumor immune infiltrates by flow cytometry indicate a 50–70% reduction in powerful immune inhibitory myeloid derived cells (MDSCs) with no changes in the total level of CD8+ and CD4+ cells. Our results show for the first time that FYN reduces anti-glioma immune responses, likely through a reduction in inhibitory MDSCs. The specific inhibition of FYN exclusively within glioma cells, could improve the efficacy of anti-glioma immunotherapies.

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