Neuroscience 2004 Teaching in Neuroscience session # 30.7 CHOLESTEROL HOMEOSTASIS FAILURE: A UNIFYING CAUSE OF SYNAPTIC DEGENERATION ? Natalia V. Koudinova,

Neuroscience 2004 Teaching in Neuroscience session # 30.7

CHOLESTEROL CHOLESTEROL HOMEOSTASIS FAILURE: HOMEOSTASIS FAILURE:

A UNIFYING CAUSE OF A UNIFYING CAUSE OF

SYNAPTIC DEGENERATION ?SYNAPTIC DEGENERATION ?Natalia V. Koudinova, Temirbolat T. Berezov Natalia V. Koudinova, Temirbolat T. Berezov

& Alexei R. Koudinov& Alexei R. Koudinov

Russia, IsraelAlso, please see slide show notes article next to this poster board

ISSN 1683-5506 / Copyright © 2004 by NV Koudinova et al. licensee Neurobiology of LipidsTo cite this article, use: Neurobiol. Lipids 3, 7 (2004), available at: http://neurobiologyoflipids.org/content/3/7/

This document is Acrobat .PDF imprint of the teaching poster slide show presented at the 34 th SFN Annual Meeting, 23-27 October 2004, San Diego, CA. To read web enhanced proceedings article and provide comments, go to: http://neurobiologyoflipids.org/content/3/7/

Cholesterol and synaptic plasticity:

Experimental system 1:Experimental system 1:

Naive albino vistar rats, acute cholesterol modulation in the hippocampal slices ex

vivoExperimental system 2:Experimental system 2:

albino vistar rats fed 2% cholesterol diet

Koudinova et al: Cholesterol failure is a unifying cause of neurodegeneration

Neurobiology of Lipids, 2004, Vol. 3, 7, http://neurobiologyoflipids.org/content/3/7

Experimental design

Incubation Incubation with cholesterol with cholesterol

acceptorsacceptors

Metabolic labeling Metabolic labeling withwith 3 3H-cholesterol H-cholesterol

or or 1414C-cholineC-choline

Electro-physiology

Immunofluorescent analysis

Efflux analysisFST catalog



Biochemical Analysis of cholesterol and phospholipid Biochemical Analysis of cholesterol and phospholipid efflux from rat hippocampal slices after incubation with efflux from rat hippocampal slices after incubation with

cyclodextrin and CSF-HDLcyclodextrin and CSF-HDL

FASEB J 2001( Science 2002 )



Cyclodextrine (cholesterol acceptor)

Fluorescent analysis of increased cholesterol efflux from rat hippocampal slices by staining with filipin

6h MCD, CA1 SR, higher x

Note cholesterol depleted SR neuronal processes (middle panel and to the

right) and pyramidal (top) and granule (bottom) cell layers in this

column of images.Navigate to our FASEBJ2001 article

at our Internet Office for better image quality and further discussion



control

cyclodextrine

B/C.B/C. Synaptic Synaptic Plasticity (LTP)Plasticity (LTP)

D. D. Presynaptic Presynaptic Mechanisms (PPF)Mechanisms (PPF)

A.A. Basic synaptic Basic synaptic function function ((I/O)I/O)

Cholesterol Cholesterol dynamicsdynamicsaffects:affects:



Cholesterol and synaptic plasticityCholesterol and synaptic plasticity

I/OI/O

PPFPPF

control

cyclodextrine

cyclodextrine

control

LTPLTP




control cyclodextrine

control

cyclodextrine

Paired PulsePaired PulseFacilitationFacilitation study study ((PPF)PPF)

NIH Alzheimer’s booklet



Increased Increased cholesterol cholesterol

Dynamics causes Dynamics causes NFT(PHF)-like NFT(PHF)-like

excessive excessive phosphorylation phosphorylation of TAU in the of TAU in the hippocampus. hippocampus.

-- see framed -- see framed panel E – it shows panel E – it shows CA3 staining with CA3 staining with monoclonal AT8 monoclonal AT8

antibody, antibody, detecting PHF-taudetecting PHF-tau

No MCD, Z1

No MCD, TAU1

MCD, AT8

MCD, TAU1

No MCD, Z1

MCD, Z1 FASEB J 2001( Science 2002 )

12



Normal Neuron

Microtubules

Neurofibrillary tangles

Degenerativeneuron

Neurofibrillary tangles (NFT)Neurofibrillary tangles (NFT)




Cholesterol-dependent change in neurofilament integrityCholesterol-dependent change in neurofilament integrityN

o M

No

M C

DC

DMM

CD

CD

FASEB J 2001CA1 Stratum Radiatum (SR) CA1SR, higher x DG, Insets: DG, higher x

FASEB J 2001

CONTROL

MCD



A staining shows no effect

of acute cholesterol

modulation on A staining

pattern_________

Slide to follow: chronic cholesterol modulation

affects A

INT

AC

T C

HO

LIN

TA

CT

CH

OL

AC

UT

E

CH

OL

A

CU

TE

C

HO

L

EF

FL

UX

, ME

FF

LU

X, M

CD

CD

FASEB J 2001

CA1SUBKoudinova et al: Cholesterol failure is a unifying cause of neurodegeneration

Neurobiol Lipids, 2004, Vol. 3, 7, http://neurobiologyoflipids.org/content/3/7

Chronic brain cholesterol homeostasis disbalance may cause Alzheimer’s like amyloid neurobiochemistry change (B-E) and

synaptic plasticity failure (F)



AA is an HDL protein constituent is an HDL protein constituent

LipoproteinLipoproteinparticleparticle

Amyloid beta

Soluble A is an apolipoprotein constituent of plasma and CSF HDL (HDL3) and CNS lipoproteins

(Koudinov et al. BBRC ‘1994, 1996, CCA’98, FASEB J ’98, many reports by others)

Chem and Life (in Russian): Jan 1998



AA in in Alzheimer’s Alzheimer’s CSF HDL CSF HDL

has has different different

distribution distribution comparedcomparedto normal to normal

CSFCSF6E10 Fluorogram Neurosci Lett 2001



Function of amyloid beta in lipid Function of amyloid beta in lipid metabolism (1996-2001)metabolism (1996-2001)

Neurochem Res ‘2000, FASEB J ‘2001

• Inhibition of cholesterol esterificationInhibition of cholesterol esterification• Inhibition of cholesterol and phospholipid synthesis in Inhibition of cholesterol and phospholipid synthesis in

liver cellsliver cells

• Increase of cholesterol and phospholipid synthesis by Increase of cholesterol and phospholipid synthesis by amyloid beta in PC12, primary neuronal cells, in fetal amyloid beta in PC12, primary neuronal cells, in fetal

rat brain, and in adult rat slicesrat brain, and in adult rat slices• Inhibition of cholesterol and phospholipid synthesis in Inhibition of cholesterol and phospholipid synthesis in

fetal rat liverfetal rat liver

Koudinov et al. BMBI ‘1996, FEBS Lett ‘1997, FASEB J ‘1998



Fluorescence microscopy

of labeled amyloid in neuronal cell culture: the mechanism of A

action on lipid synthesis remain

obscure.

Neurochem Res ‘2000



Amyloid beta, LTP and hippocampal Amyloid beta, LTP and hippocampal lipid synthesislipid synthesis




CONDITIONS HAVING CHANGED CONDITIONS HAVING CHANGED AMYLOID AMYLOID and/or TAU NEUROCHEMISTRY and/or TAU NEUROCHEMISTRY


Alzheimer’s disease (AAlzheimer’s disease (A/TAU)/TAU)

Cardiovascular disease (ACardiovascular disease (A))Down syndrome (ADown syndrome (A))

Frontotemporal dementia (TAU) Frontotemporal dementia (TAU) Inclusion-body myositis (AInclusion-body myositis (A, TAU), TAU)ALS (TAU, AALS (TAU, A in brain/skin) in brain/skin)Parkinson disease (TAU)Parkinson disease (TAU)________________________________________________________

Normal early postnatal development Normal early postnatal development (TAU, not known for A(TAU, not known for A))Normal aging (ANormal aging (A))



What about different neurodegenerative diseases ?

* ** *

Is there a relevance

of this part of the cascade to other

neurodegenerative diseases ?

---------

If so, is there a neurodegeneration

specificity?

In Parkinson disease, for example, synaptic function

break may be due to a primary break of the

forebrain dopamine system

This scheme was first developed by us to explain the pathogenesis of Alzheimer’s



Free open access at:

http://neurobiologyoflipids.org

Cholesterol and Alzheimer’sKoudinova et al: Cholesterol failure is a unifying cause of neurodegeneration


Cholesterol and Down syndromeCholesterol and Down syndrome

CCholesterol esterification abnormalityholesterol esterification abnormality in DS in DS patientspatients

SSpecific pathway of the liver sterol regulatory pecific pathway of the liver sterol regulatory element binding protein (SREBP) activation with element binding protein (SREBP) activation with

sterol-independent maturation of SREBP-1sterol-independent maturation of SREBP-1

HHigh circulating and tissue cholesterol levels in the igh circulating and tissue cholesterol levels in the fetusesfetuses

Lacko AG et al.. Clin Chim Acta. 132, 133 (1983).Diomede L, et al. BBRC. 260, 503 (1999).



NMJ and ANMJ and AAPP metabolismAPP metabolism

Askanas V, et al. Neurosci Lett 1992 143: 96-100.

1992 - the first demonstration of the amyloid precursor 1992 - the first demonstration of the amyloid precursor protein (APP) and Aprotein (APP) and A sequences concentrated sequences concentrated

postsynaptically at human neuro-muscular junctions postsynaptically at human neuro-muscular junctions (NMJs). (NMJs).

Conclusion: APP may have a role in normal junction biology Conclusion: APP may have a role in normal junction biology and possibly in some diseases affecting NMJs.and possibly in some diseases affecting NMJs.

Torroja L, et al. J Neurosci 1999 19: 7793-803.

1999 - The Drosophila APP homolog promotes 1999 - The Drosophila APP homolog promotes synapse differentiation at the neuromuscular synapse differentiation at the neuromuscular

junction.junction.



A restores hippocampal

LTP,

cholesterol synthesis inhibition

absorbs the restorative

action of the peptide



Cholesterol and Inclusion-body Cholesterol and Inclusion-body myositis (IBM)myositis (IBM)

IBM vacuolated muscle fibers (VMF) IBM vacuolated muscle fibers (VMF) had strong LDLR- and VLDLR-immunoreactive had strong LDLR- and VLDLR-immunoreactive inclusions, which colocalized with Abeta, APOE, inclusions, which colocalized with Abeta, APOE,

p-tau, and free cholesterol. p-tau, and free cholesterol.

CONCLUSIONS:CONCLUSIONS:

Abnormal accumulation of LDLR, VLDLR, LRP, and Abnormal accumulation of LDLR, VLDLR, LRP, and cholesterol within IBM vacuolated muscle fibers cholesterol within IBM vacuolated muscle fibers suggests novel roles for them in the pathogenesis suggests novel roles for them in the pathogenesis

of the inclusion-body myositis.of the inclusion-body myositis.

Jaworska-Wilczynska et al. Neurology 2002; 58: 438.



Apo E and neuromuscular diseaseApo E and neuromuscular disease

ApoE e4 association with an increased risk for ApoE e4 association with an increased risk for developing and faster progression of certain developing and faster progression of certain

neuromuscular diseases, while neuromuscular diseases, while

ApoE e2 may have the protective effect and is ApoE e2 may have the protective effect and is associated with better prognosis for neuromuscular associated with better prognosis for neuromuscular

diseases such as motor neuron disease. diseases such as motor neuron disease.

The effect of various APOE alleles on neuromuscular The effect of various APOE alleles on neuromuscular diseases therefore parallels their influence on diseases therefore parallels their influence on

central nervous system diseases.central nervous system diseases.

Bedlack, Strittmatter, Morgenlander. Arch Neurol 2000 57: 1561-5.



Cholesterol and Receptor functionCholesterol and Receptor functionnAChR nAChR

Reviewed by Naguib et al, Anesthesiology 2002Reviewed by Naguib et al, Anesthesiology 2002

GABAGABAAASooksawate and Simmond, Neuropharmacology, 2001, Br J Pharm 2001Sooksawate and Simmond, Neuropharmacology, 2001, Br J Pharm 2001

5HT1A5HT1AScanlon, Williams, Schloss. Biochemistry 2001Scanlon, Williams, Schloss. Biochemistry 2001

Scalzitti et al. Society for Neurosci Abstract 2003, Scalzitti et al. Society for Neurosci Abstract 2003, available at Neurobiology of Lipids, Vol. 2, 3available at Neurobiology of Lipids, Vol. 2, 3

GlyRGlyRLaube. Society for Neurosci Abstract 2003, Laube. Society for Neurosci Abstract 2003, available at Neurobiology of Lipids, Vol. 2, 3available at Neurobiology of Lipids, Vol. 2, 3

Excitatory Amino Acid TransportersExcitatory Amino Acid TransportersButchbach, Guo, Lin. Society for Neurosci Abstract 2003, Butchbach, Guo, Lin. Society for Neurosci Abstract 2003,

available at Neurobiology of Lipids, Vol. 2, 3available at Neurobiology of Lipids, Vol. 2, 3http://neurobiologyoflipids.org/content/2/3http://neurobiologyoflipids.org/content/2/3



TAKE HOME MESSAGETAKE HOME MESSAGE

Cholesterol itself plays essential role in the mechanisms of synaptic function, plasticity and

neurodegeneration

Central neurodegeneration features in a

number of neurodegenerative diseases

may represent functional consequences

of abnormal neural cholesterol

misregulation

KOUDINOVSKoudinova et al: Cholesterol failure is a unifying cause of neurodegeneration


Documents

Neuroscience 2004 Teaching in Neuroscience session # 30.7 CHOLESTEROL HOMEOSTASIS FAILURE: A UNIFYING CAUSE OF SYNAPTIC DEGENERATION ? Natalia V. Koudinova,