Liver-specific γ-glutamyl carboxylase-deficient mice display bleeding diathesis and short life span

Kotaro Azuma; Tohru Tsukui; Kazuhiro Ikeda; Sachiko Shiba; Kimie Nakagawa; Toshio Okano; Tomohiko Urano; Kuniko Horie-Inoue; Yasuyoshi Ouchi; Masahito Ikawa; Satoshi Inoue

doi:10.1371/journal.pone.0088643

Liver-specific γ-glutamyl carboxylase-deficient mice display bleeding diathesis and short life span

PLoS One. 2014 Feb 10;9(2):e88643. doi: 10.1371/journal.pone.0088643. eCollection 2014.

Authors

Affiliations

¹ Department of Geriatric Medicine, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
² Division of Gene Regulation and Signal Transduction, Research Center for Genomic Medicine, Saitama Medical University, Hidaka, Saitama, Japan.
³ Department of Hygienic Sciences, Kobe Pharmaceutical University, Kobe, Hyogo, Japan.
⁴ Department of Anti-Aging Medicine, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
⁵ Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, Japan.
⁶ Department of Geriatric Medicine, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, Japan ; Division of Gene Regulation and Signal Transduction, Research Center for Genomic Medicine, Saitama Medical University, Hidaka, Saitama, Japan ; Department of Anti-Aging Medicine, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.

Abstract

Vitamin K is a fat-soluble vitamin that plays important roles in blood coagulation and bone metabolism. One of its functions is as a co-factor for γ-glutamyl carboxylase (Ggcx). Conventional knockout of Ggcx causes death shortly after birth in homozygous mice. We created Ggcx-floxed mice by inserting loxP sequences at the sites flanking exon 6 of Ggcx. By mating these mice with albumin-Cre mice, we generated Ggcx-deficient mice specifically in hepatocytes (Ggcx(Δliver/Δliver) mice). In contrast to conventional Ggcx knockout mice, Ggcx(Δliver/Δliver) mice had very low activity of Ggcx in the liver and survived several weeks after birth. Furthermore, compared with heterozygous mice (Ggcx(+/Δliver) ), Ggcx(Δliver/Δliver) mice had shorter life spans. Ggcx(Δliver/Δliver) mice displayed bleeding diathesis, which was accompanied by decreased activity of coagulation factors II and IX. Ggcx-floxed mice can prove useful in examining Ggcx functions in vivo.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Carbon-Carbon Ligases / deficiency*
Carbon-Carbon Ligases / metabolism
Gene Deletion
Hemorrhagic Disorders / enzymology*
Hemorrhagic Disorders / pathology*
Hepatocytes / enzymology
Hepatocytes / pathology
Integrases / metabolism
Liver / enzymology*
Liver / pathology
Longevity*
Mice
Mice, Knockout
Organ Specificity

Substances

Cre recombinase
Integrases
Carbon-Carbon Ligases
glutamyl carboxylase

Grants and funding

This work was supported by grants from Cell Innovation Program, Grants-in-Aid, and Support Project of Strategic Research Center in Private Universities from the Ministry of Education, Culture, Sports, Science and Technology, Grants-in-Aid from the Ministry of Health, Labour, and Welfare, and the Program for Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.