Susceptibility of transgenic mice expressing human apolipoprotein E to closed head injury: The allele E3 is neuroprotective whereas E4 increases fatalities

doi:10.1016/S0306-4522(00)00438-3

Neuroscience

Volume 101, Issue 4, 30 November 2000, Pages 879-884

https://doi.org/10.1016/S0306-4522(00)00438-3 Get rights and content

Abstract

Apolipoprotein E, the major brain lipid-binding protein, is expressed in humans as three common isoforms (E2, E3 and E4). Previous studies revealed that the allele apolipoprotein E4 is a major genetic risk factor of Alzheimer’s disease and that traumatic brain injury is associated with increased risk for developing this disease. Furthermore, it has been suggested that the effects of traumatic head injury and apolipoprotein E4 in Alzheimer’s disease are synergistic. To test the hypothesis that the apolipoprotein E genotype affects susceptibility to brain injury, we subjected transgenic mice, expressing either human apolipoprotein E3 or human apolipoprotein E4 on a null mouse apolipoprotein E background and apolipoprotein E-deficient knockouts, to closed head injury and compared mortality, neurological recovery and the extent of brain damage of the survivors. More than 50% of the transgenic mice expressing human apolipoprotein E4 died following closed head injury, whereas only half as many of the transgenic mice expressing human apolipoprotein E3, and of the control and apolipoprotein E-deficient mice died during this period (P<0.02). A neurological severity score used for clinical assessment of the surviving mice up to 11 days after closed head injury revealed that the four mouse groups displayed similar severity of damage at 1 h following injury. At three and 11 days post-injury, however, the neurological severity scores of the transgenic mice expressing human apolipoprotein E3 were significantly lower than those of the other three groups whose scores were similar, indicating better recovery of the transgenic mice expressing human apolipoprotein E3. Histopathological examination of the mice performed 11 days post-injury revealed, consistent with the above neurological results, that the size of the damaged brain area of the transgenic mice expressing human apolipoprotein E3 was smaller than that of the other head-injured groups.

These findings show that transgenic mice expressing human apolipoprotein E4 are more susceptible than those expressing apolipoprotein E3 to closed head injury. We suggest that this effect is due to both a protective effect of apolipoprotein E3 and an apolipoprotein E4-related pathological function.

Section snippets

Animals

Human apoE3 and apoE4 transgenic mice were generated on an apoE-deficient C57BL/6J background utilizing human apoE3 and apoE4 transgenic constructs as previously reported.⁵³ Accordingly, cosmid libraries were constructed from lymphoblasts of humans known to be homozygous carriers for apoE3 or apoE4, after which fragments containing human regulatory sequences and the coding sequences for human apoE were used to produce the transgenic mice. The experiments were performed with the apoE3-453 and

Results

The susceptibilities of the apoE3 and apoE4 transgenic mice and of apoE-deficient and normal control mice to CHI were assessed by measurements of their neurological status during 11 days following head injury and by monitoring the number of mice that died in each group.

As shown in Table 2, four control, six apoE-deficient and five apoE3 transgenic mice died within 11 days following CHI, which translates approximately to the same percentage of the total mice for each of these groups (i.e. 27%,

Discussion

This study revealed that mice transgenic for human apoE4 are more susceptible to CHI than human apoE3 transgenic mice. Furthermore, neurological and histological analysis revealed that the apoE3 transgenic mice recovered more rapidly from CHI than control, apoE-deficient and apoE4 transgenic mice and that the infarct size of the injured apoE3 mice was significantly smaller than that of the other mouse groups. In contrast, analysis of the fatalities following CHI revealed that a similar fraction

Conclusions

This study has revealed that human apoE3 transgenic mice are less susceptible than human apoE4 transgenic mice to CHI and that this effect is due both to increased vulnerability of the apoE4 transgenic mice, manifested in increased fatality, and to more effective recovery of the apoE3 mice.

Acknowledgements

We thank Duke University and Glaxo Wellcome for kindly providing the transgenic mice. This work was supported partly by grants from the Joint German and Israeli Research Projects sponsored by the German and Israeli Ministries of Science (grant no. 1626), from the Harry Stern National Center for Alzheimer’s Disease and Related Disorders, from the Jo and Inez Eichenbaum Foundation and from the Revah-Kabelli Fund. D.M. Michaelson is the incumbent of the Myriam Lebach Chair in Molecular

References (54)

Y. Chen et al.
Motor and cognitive deficits in apolipoprotein E-deficient mice after closed head injury
Neuroscience
(1997)
S.M. Gentleman et al.
Molecular pathology of head trauma: altered βAPP metabolism and the aetiology of Alzheimer’s disease
Prog. Brain Res.
(1993)
M.J. Ignatius et al.
Apolipoprotein E in nerve injury and repair
Prog. Brain Res.
(1987)
L. Lomnitski et al.
Reduced levels of antioxidants in brains of apolipoprotein E-deficient mice following closed head injury
Pharmac. Biochem. Behav.
(1997)
E. Masliah et al.
Neurodegeneration and cognitive impairment in apoE-deficient mice is ameliorated by infusion of recombinant apoE
Brain Res.
(1997)
M.F. Newman et al.
Predictors of cognitive decline after cardiac operation
Ann. Thorac. Surg.
(1995)
J. Poirier
Apolipoprotein E in animal models of CNS injury and in Alzheimer’s disease
Trends neurol. Sci.
(1994)
W.J. Strittmatter et al.
Hypothesis: microtubule instability and paired helical filament formation in the Alzheimer disease brain are related to apolipoprotein E genotype
Expl Neurol.
(1994)
I. Veinbergs et al.
Altered long-term potentiation in the hippocampus of apolipoprotein E-deficient mice
Neurosci. Lett.
(1998)
I. Veinbergs et al.
Differential neurotrophic effects of apolipoprotein E in aged transgenic mice
Neurosci. Lett.
(1999)

P.R. Wenham et al.

Apolipoprotein E genotyping by one-stage PCR (letter)

Lancet

(1991)

T Wisniewski et al.

Apolipoprotein E: a pathological chaperone protein in patients with cerebral and systemic amyloid

Neurosci. Lett.

(1992)

P.-T. Xu et al.

Specific regional transcription of apolipoprotein E in human brain neurons

Am. J. Path.

(1999)

P.-T. Xu et al.

Human apolipoprotein E2, E3 and E4 isoform-specific transgenic mice: human-like pattern of glial and neuronal immunoreactivity in central nervous system not observed in wild-type mice

Neurobiol. Dis.

(1996)

H. Basun et al.

Apolipoprotein E polymorphism and stroke in a population sample aged 75 years or more

Stroke

(1996)

S. Bellosta et al.

Stable expression and secretion of apolipoprotein E3 and E4 in mouse neuroblastoma cells produces differential effects on neurite outgrowth

J. biol. Chem.

(1995)

J.K. Boyles et al.

Apolipoprotein E associated with astrocytic glia of the central nervous system and with nonmyelinating glia of the peripheral nervous system

J. clin. Invest.

(1985)

M. Buttini et al.

Expression of human apolipoprotein E3 or E4 in the brains of apoE-/- mice: isoform-specific effects on neurodegeneration

J. Neurosci.

(1999)

J. Chapman et al.

A simple and efficient method for apolipoprotein E genotype determination

Neurology

(1996)

J. Chapman et al.

Preliminary observations on APOE E4 allele and progression of disability in multiple sclerosis

J. Am. med. Ass.

(1999)

E.H. Corder et al.

Gene dose of apolipoprotein E type 4 allele and the risk of Alzheimer’s disease in late onset families

Science

(1993)

M.A. Elshourbagy et al.

Apolipoprotein E mRNA is abundant in the brain and adrenals as well as in the liver and is present in other peripheral tissues of rats and marmosets

Proc. natn. Acad. Sci. USA

(1985)

L.R. French et al.

A case–control study of dementia of the Alzheimer type

J. Epidemiol.

(1991)

I. Genis et al.

Tau hyperphosphorylation in apolipoprotein E-deficient and control mice following closed head injury

J. Neurosci. Res.

(2000)

S.-H. Han et al.

Apolipoprotein E is localized to the cytoplasm of human cortical neurons: a light and electron microscopic study

J. Neuropath. exp. Neurol.

(1994)

Z.-S. Ji et al.

Differential cellular accumulation/retention of apolipoprotein E mediated by cell surface heparin sulfate proteoglycans. Apolipoproteins E3 and E2 greater than E4

J. biol. Chem.

(1998)

C. Knouff et al.

ApoE structure determines VLDL clearance and atherosclerosis risk in mice

J. clin. Invest.

(1999)

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^†: Permanent address: Israel Institute for Biological Research, P.O. Box 19, Ness-Ziona, Israel.

^‡: The first two authors contributed equally to this work.

View full text

Susceptibility of transgenic mice expressing human apolipoprotein E to closed head injury: The allele E3 is neuroprotective whereas E4 increases fatalities

Abstract

Section snippets

Animals

Results

Discussion

Conclusions

Acknowledgements

Neuroscience

Prog. Brain Res.

Prog. Brain Res.

Pharmac. Biochem. Behav.

Brain Res.

Ann. Thorac. Surg.

Trends neurol. Sci.

Expl Neurol.

Neurosci. Lett.

Neurosci. Lett.

Lancet

Neurosci. Lett.

Am. J. Path.

Neurobiol. Dis.

Apolipoprotein E polymorphism and stroke in a population sample aged 75 years or more

Stroke

Stable expression and secretion of apolipoprotein E3 and E4 in mouse neuroblastoma cells produces differential effects on neurite outgrowth

J. biol. Chem.

Apolipoprotein E associated with astrocytic glia of the central nervous system and with nonmyelinating glia of the peripheral nervous system

J. clin. Invest.

Expression of human apolipoprotein E3 or E4 in the brains of apoE-/- mice: isoform-specific effects on neurodegeneration

J. Neurosci.

A simple and efficient method for apolipoprotein E genotype determination

Neurology

Preliminary observations on APOE E4 allele and progression of disability in multiple sclerosis

J. Am. med. Ass.

Gene dose of apolipoprotein E type 4 allele and the risk of Alzheimer’s disease in late onset families

Science

Apolipoprotein E mRNA is abundant in the brain and adrenals as well as in the liver and is present in other peripheral tissues of rats and marmosets

Proc. natn. Acad. Sci. USA

A case–control study of dementia of the Alzheimer type

J. Epidemiol.

Tau hyperphosphorylation in apolipoprotein E-deficient and control mice following closed head injury

J. Neurosci. Res.

Apolipoprotein E is localized to the cytoplasm of human cortical neurons: a light and electron microscopic study

J. Neuropath. exp. Neurol.

Differential cellular accumulation/retention of apolipoprotein E mediated by cell surface heparin sulfate proteoglycans. Apolipoproteins E3 and E2 greater than E4

J. biol. Chem.

ApoE structure determines VLDL clearance and atherosclerosis risk in mice

J. clin. Invest.