Complete hepatic regeneration after somatic deletion of an albumin-plasminogen activator transgene

doi:10.1016/0092-8674(91)90615-6

Cell

Volume 66, Issue 2, 26 July 1991, Pages 245-256

https://doi.org/10.1016/0092-8674(91)90615-6 Get rights and content

Abstract

We previously demonstrated that expression of an albumin-urokinase-type plasminogen activator (Alb-uPA) fusion construct in transgenic mice resulted in elevated plasma uPA concentration, hypofibrinogenemia, and neonatal hemorrhaging. Two lines of Alb-uPA mice were established in which only one half of the transgenic pups died at birth; surprisingly, plasma uPA concentrations in survivors gradually returned to normal by 2 months of age. The basis for this phenomenon is DNA rearrangement within hepatocytes that affects the transgene tandem array and abolishes transgene expression. Transgene-deficient cells selectively proliferate relative to surrounding liver, and this process culminates in replacement of the entire liver by clonal hepatic nodules derived from transgene-deficient progenitor cells. In some cases as few as two nodules can reconstitute over 90% of liver mass, highlighting the remarkable regenerative capacity of individual liver cells.

References (46)

W.Y. Brodsky et al.
Cell polyploidy: its relation to tissue growth and function
Int. Rev. Cytol.
(1977)
F.V. Chisari et al.
Molecular pathogenesis of hepatocellular carcinoma in hepatitis B virus transgenic mice
Cell
(1989)
J.L. Heckel et al.
Neonatal bleeding in transgenic mice expressing urokinase-type plasminogen activator
Cell
(1990)
D.R. Higgs et al.
A review of the molecular genetics of the human globin gene cluster
Blood
(1989)
C. Jhappan et al.
TGFα overexpression in transgenic mice induces liver neoplasia and abnormal development of the mammary gland and pancreas
Cell
(1990)
M.A. Lehrman et al.
Duplication of seven exons in LDL receptor gene caused by Alu-Alu recombination in a subject with familial hypercholesterolemia
Cell
(1987)
R.D. Palmiter et al.
Differential regulation of metal loth ionein-thymidine kinase fusion genes in transgenic mice and their offspring
Cell
(1982)
R.D. Palmiter et al.
Transmission distortion and mosaicism in an unusual transgenic mouse pedigree
Cell
(1984)
R.D. Palmiter et al.
Cell lineage ablation in transgenic mice by cell-specific expression of a toxin gene
Cell
(1987)
E.P. Sandgren et al.
Overexpression of TGFa in transgenic mice: induction of epithelial hyperplasia, pancreatic metaplasia, and carcinoma of the breast
Cell
(1990)

R. Soda et al.

Liver endothelium and not hepatocytes or Kupffer cells have transferrin receptors

Blood

(1984)

P.H. Yen et al.

Frequent deletions of the human X chromosome distal short arm result from recombination between low copy repetitive elements

Cell

(1990)

E. Bakker et al.

Germline mosaicism and Duchenne muscular dystrophy mutations

Nature

(1987)

R.R. Behringer et al.

Dwarf mice produced by genetic ablation of growth hormoneexpressing cells

Genes Dev.

(1988)

R.E. Braun et al.

Infertility in male transgenic mice: disruption of sperm development by HSV-tk expression in post meiotic germ cells

Biol. Reprod.

(1990)

R.L. Brinster et al.

Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs

J.A. Carlson et al.

Accumulation of PiZ α₁-antitrypsin causes liver damage in transgenic mice

J. Clin. Invest.

(1989)

F.V. Chisari et al.

A transgenic mouse model of the chronic hepatitis B surface antigen carrier state

Science

(1985)

F.V. Chisari et al.

Structural and pathological effects of synthesis of hepatitis B virus large envelope polypeptide in transgenic mice

D. Collen et al.

Fibrinolysis and the control of hemostasis

K. Danø et al.

Plasminogen activators, tissue degradation, and cancer

Adv. Cancer Res.

(1985)

B.T. Darras et al.

A partial deletion of the muscular dystrophy gene transmitted twice by an unaffected male

Nature

(1987)

S.J.F. Degen et al.

Characterization of the cDNA coding for mouse plasminogen and localization of the gene to mouse chromosome 17

Genomics

(1990)

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Cell

ArticleComplete hepatic regeneration after somatic deletion of an albumin-plasminogen activator transgene

Abstract

Int. Rev. Cytol.

Cell

Cell

Blood

Cell

Cell

Cell

Cell

Cell

Cell

Blood

Cell

Germline mosaicism and Duchenne muscular dystrophy mutations

Nature

Dwarf mice produced by genetic ablation of growth hormoneexpressing cells

Genes Dev.

Infertility in male transgenic mice: disruption of sperm development by HSV-tk expression in post meiotic germ cells

Biol. Reprod.

Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs

Accumulation of PiZ α1-antitrypsin causes liver damage in transgenic mice

J. Clin. Invest.

A transgenic mouse model of the chronic hepatitis B surface antigen carrier state

Science

Structural and pathological effects of synthesis of hepatitis B virus large envelope polypeptide in transgenic mice

Fibrinolysis and the control of hemostasis

Plasminogen activators, tissue degradation, and cancer

Adv. Cancer Res.

A partial deletion of the muscular dystrophy gene transmitted twice by an unaffected male

Nature

Characterization of the cDNA coding for mouse plasminogen and localization of the gene to mouse chromosome 17

Genomics

Article
Complete hepatic regeneration after somatic deletion of an albumin-plasminogen activator transgene

Accumulation of PiZ α₁-antitrypsin causes liver damage in transgenic mice