Peroxisome Proliferator-activated Receptor (PPAR)-γ Modifies Aβ Neurotoxin-induced Electrophysiological Alterations in Rat Primary Cultured Hippocampal Neurons

Document Type : Research article

Authors

1 Neuroscience Research Center, Department of Physiology, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran, Iran.

2 Neuroscience Research Center, Department of Physiology, School of Medical, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Abstract

Alzheimer’s disease (AD) is undoubtedly one of the serious and growing public health challenges in the world today. There is an unmet need for new and effective preventative and therapeutic treatment approaches for AD, particularly at early stages of the disease. However, the underlying mechanism against Aβ-induced electrophysiological alteration in cultured hippocampal pyramidal neurons  is still not fully understood. This study investigated  the impacts of activation and inhibition of PPAR-γ/δ on the Aβ-induced functional toxicity, which occured before cell death, using patch clamp technique. Findings demonstrated that Aβ treatment alone altered the normal electrophysiological properties and reduced the Ca2+ channel currents in primary cultured hippocampal pyramidal neurons without any major changes either in cell structure, as evidenced by electron microscope examination, or cell viability. Rosiglitazone (30 μM), a potent PPAR-γ activator, when co-treated with Aβ (100 nM) prevented almost completely the induction of function toxicity of Aβ, as evidentiated by restored normal appearing electrophysiological properties. Inhibition of PPAR- γ/δ by FH535 (15 μM), an inhibitor of both Wnt/beta-catenin signaling and PPAR- γ and δ activity, when applied in combination of Aβ not only worsen the toxic electrophysiological effects of Aβ on firing frequency, membrane resistance and cell viability, but also even preserved the suppressive effect of Aβ on Ca2+ channel current when compared to control condition. Overall, these findings suggest that PPAR-γ activation could be a potential candidate to prevent the functional changes induced by low concentration of Aβ which may possibly occur in neurons during early stages of AD.

Keywords

Main Subjects


triepens N, Scheef L, Wind A, Popp J, Spottke A,
Cooper-Mahkorn D, Suliman H, Wagner M, Schild

HH and Jessen F. Volume loss of the medial temporal

lobe structures in subjective memory impairment.

Dement. Geriatr. Cogn. Disord
. (2010) 29: 75-81.
Hardy J and Selkoe DJ. The amyloid hypothesis of

alzheimer’s disease: progress and problems on the

road to therapeutics.
Science (2002) 297: 353-6.
Magdesian MH, Carvalho MM, Mendes FA, Saraiva

LM, Juliano MA, Juliano L, Garcia-Abreu J and

Ferreira ST. Amyloid-beta binds to the extracellular

cysteine-rich domain of Frizzled and inhibits Wnt/

beta-catenin signaling.
J. Biol. Chem. (2008) 283:
9359-68.

Dunn L, Allen GF, Mamais A, Ling H, Li A, Duberley

KE, Hargreaves IP, Pope S, Holton JL, Lees A, Heales

SJ
and Bandopadhyay R. Dysregulation of glucose
metabolism is an early event in sporadic parkinson’s

disease.
Neurobiol. Aging (2014) 35: 1111-5.
Ontiveros-Torres MÁ, Labra-Barrios ML, Díaz-

Cintra S, Vázquez-Aguilar A, Moreno-Campuzano

S, Flores-Rodríguez P, Luna-Herrera C, Mena R,

Perry G, Florán-Garduño B, Luna-Muñoz J and

Luna-Arias JP. Fibrillar amyloid-β accumulation

triggers an inflammatory mechanism leading to

hyperphosphorylation of the carboxyl-terminal end of

tau polypeptide in the hippocampal formation of the

3×Tg-AD transgenic mouse.
J. Alzheimers Dis. (2016)
52: 243-69.

Fernandez-Perez EJ, Peters C and Aguayo LG.

Membrane damage induced by amyloid beta and a

potential link with neuroinflammation.
Curr. Pharm.
Des.
(2016) 22: 1295-304.
Ferré P. The biology of peroxisome proliferator-

activated receptors: relationship with lipidmetabolism

and insulin sensitivity.
Diabetes (2004) 53 (Suppl 1):
S43-50.

Jiang Q, Heneka M and Landreth GE. The role of

peroxisome proliferator-activated receptor-gamma

(PPARgamma) in
alzheimer’s disease: therapeutic
implications.
CNS Drugs (2008) 22: 1-14.
Pedersen WA, McMillan PJ, Kulstad JJ, Leverenz JB,

Craft S and Haynatzki GR. Rosiglitazone attenuates

learning and memory deficits in Tg2576 alzheimer

mice.
Exp. Neurol. (2006) 199: 265–73.
O′Reilly JA and Lynch M. Rosiglitazone improves

spatial memory and decreases insoluble Aβ(1-42) in

APP/PS1 mice.
J. Neuroimmune Pharmacol. (2012)7: 140-4.
Xu S, Liu G, Bao X, Wu J, Li S, Zheng B, Anwyl

R and Wang Q. Rosiglitazone prevents amyloid-β

oligomer-induced impairment of synapse formation

and plasticity via increasing dendrite and spine

mitochondrial number.
J. Alzheimers Dis. (2014) 39:
239-51.

Miller BW, Willett KC and Desilets AR. Rosiglitazone

and pioglitazone for the treatment of alzheimer′s

disease.
Ann. Pharmacother. (2011) 45: 1416-24.
Heneka MT, Reyes-Irisarri E, Hüll M and Kummer

MP. Impact and therapeutic potential of PPARs in

alzheimer′s disease.
Curr. Neuropharmacol. (2011)
9: 643-50.

Inestrosa N, De Ferrari GV, Garrido JL, Alvarez A,

Olivares GH, Barría MI, Bronfman M and Chacón

MA. Wnt signaling involvement in beta-amyloid-

dependent neurodegeneration.
Neurochem. Int. (2002)
41: 341-4.

Inestrosa NC, Godoy JA, Quintanilla RA, Koenig

CS and Bronfman M. Peroxisome proliferator-

activated receptor gamma is expressed in hippocampal

neurons and its activation prevents beta-amyloid

neurodegeneration: role of Wnt signalling.
Exp. Cell
Res.
(2005) 304: 91-104.
Kaundal RK and Sharma SS. Peroxisome proliferator-

activated receptor gamma agonists as neuroprotective

agents.
Drug News Perspect. (2010) 23: 241-56.
Coombs GS, Covey TM and Virshup DM. Wnt

signaling in development disease and translational

medicine.
Curr. Drug Targets (2008) 9: 513–31.
Inestrosa NC and Varela-Nallar L. Wnt signaling in

thenervous system andin alzheimer’s disease.
J. Mol.
Cell Biol.
(2014) 6: 64-74.
Oliva CA, Vargas JY and Inestrosa NC. Wnts in

adult brain: from synaptic plasticity to cognitive

deficiencies.
Front. Cell Neurosci. (2013) 7: 224.
Cerpa W, Gambrill A, Inestrosa NC and Barria A.

Regulation of NMDA-receptor synaptic transmission

by Wnt signaling.
J. Neurosci. (2011) 31: 9466–71.
Zhang GL, Zhang J, Li SF, Lei L, Xie HY, Deng F,

Feng JC and Qi JS. Wnt-5a prevents Aβ-induced

deficits in long-term potentiation and spatial memory

in rats.
Physiol. Behav. (2015) 149: 95-100.
Caracci MO, Ávila ME and De Ferrari GV. Synaptic

Wnt/GSK3β signaling Hub in autism.
Neural Plast.
(2016) 2016: 10.

Inestrosa NC and Toledo EM. The role of Wnt signaling

in neuronal dysfunction in alzheimer’s disease.
Mol.
Neurodegener.
(2008) 3: 9.
Alvarez AR, Godoy JA, Mullendorff K, Olivares GH,

Bronfman M and Inestrosa NC. Wnt-3a overcomes

beta-amyloid toxicity in rat hippocampal neurons.

Exp. Cell Res.
(2004) 297: 186-96.
Hayashi Y, Hirotsu T, Iwata R, Kage-Nakadai

E and Kunitomo H. A trophic role for Wnt-Ror

kinase signaling during developmental pruning in

Caenorhabditis elegans.
Nat. Neurosci. (2009) 12:
981-7.

(11)

(12)

(13)

(14)

(15)

(16)

(17)

(18)

(19)

(20)

(21)

(22)

(23)

(24)

(25)

Acknowledgement

This research was supported by Neuroscience

Research Centers of Shahid Beheshti Uinversity

of Medical Sciences and Baqiyatallah Uinversity

of Medical Sciences.

References
 
Bahrami F et al. / IJPR (2019), 18 (3): 1403-14181416
Iida J, Dorchak J, Lehman JR, Clancy R and Luo

C. FH535 inhibited migration and growth of breast

cancer cells.
PLoS One (2012) 7: e44418.
Vilchez V, Turcios L, Marti F and Gedaly R. Targeting

Wnt/β-catenin pathway in hepatocellular carcinoma

treatment.
World J. Gastroenterol. (2016) 22: 823-32 .
Brewer GJ, Torricelli JR, Evege EK and Price PJ.

Optimized survival of hippocampal neurons in B27-

supplemented Neurobasal, a new serum-free medium

combination.
J. Neurosci Res. (1993) 35: 567-76.
Bahrami F and Janahmadi M. Antibiotic supplements

affect electrophysiological properties and excitability

of rat hippocampal pyramidal neurons in primary

culture.
Iran. Biomed. J. (2013) 17: 101-6.
Bahrami F, Yousefpour M, Mehrani H, Golmanesh L

and Sadraee SH. A Type of cell death and the role of

acetylcholinesterase activity in neurotoxicity induced

by paraoxon in cultured rat hippocampal neurons.
Acta
Biol. Hung
. (2009) 60: 1-13.
Handeli S and Simon JA. A small-molecule inhibitor

of Tcf/beta-catenin signaling down-regulates

PPARgamma and PPARdelta activities.
Mol. Cancer
Ther.
(2008) 7: 521-9.
Kanerva L and Verkkala E. Electron microscopy and

immunohistochemistry of toxic and allergic effects of

methylmethacrylate on the skin.
Arch. Toxicol. Suppl.
(1986) 9: 456-9.

Saggu SK, Chotaliya HP, Blumbergs PC and Casson

RJ. Wallerian-like axonal degeneration in the optic

nerve after excitotoxic retinal insult: an ultrastructural

study.
BMC Neurosci. (2010) 11: 97.
Caricasole A, Copani A, Caruso A, Caraci F, Iacovelli

L, Sortino MA, Terstappen GC and Nicoletti F. The

Wnt pathway, cell-cycle activation and beta-amyloid:

Novel therapeutic strategies in alzheimer’s disease?

Trends Pharmacol. Sci.
(2003) 24: 233-8.
De Ferrari GV, Chacón MA, Barría MI, Garrido

JL, Godoy JA, Olivares G, Reyes AE, Alvarez A,

Bronfman M and Inestrosa NC. Activation of Wnt

signaling rescues neurodegeneration and behavioral

impairments induced by beta-amyloid fibrils.
Mol.
Psychiatry
(2003) 8: 195-208.
Tu S, Okamoto S, Lipton SA and Xu H. Oligomeric

Aβ-induced synaptic dysfunction in alzheimer’s

disease.
Mol. Neurodegener. (2014) 14: 9-48.
Carrillo-Mora P, Luna R and Colín-Barenque L.

Amyloid beta: multiple mechanisms of toxicity and

only some protective effects?
Oxid. Med. Cell. Longev.
(2014) 2014: 15.

Lambert MP, Barlow AK, Chromy BA, Edwards C and

Freed R. Diffusible, nonfribrillar ligands derived from

1-42 are potent central nervous system neurotoxins
Proc. Natl. Acad. Sci. USA
(1988) 95: 6448-53.
Wang HW, Pasternak JF, Kuo H, Ristic H, Lambert

MP, Chromy B, Viola KL, Klein WL, Stine WB,

Krafft GA and Trommer BL. Soluble oligomers of beta

amyloid (1–42) inhibit long-term potentiation but not

long-term depression in rat dentate gyrus.
Brain Res.
(2002) 924: 133–40.

Wang Y, Zhang G, Zhou H, Barakat A and Querfurth

H. Opposite effects of low and high doses of Abeta42

on electrical network and neuronal excitability in the

rat prefrontal cortex.
PLoS One (2009) 4: e8366.
Varghese K, Molnar P, Das M, Bhargava N, Lambert

S, Kindy MS and Hickman JJ. A new target for

amyloid beta toxicity validated by standard and high-

throughput electrophysiology.
PLoS One (2010) 5:
e8643.

Hong-Qi Y, Zhi-Kun S and Sheng-Di C. Current

advances in the treatment of alzheimer’s disease:

focused on considerations targeting Aβ and tau.
Transl.
Neurodegener.
(2012) 1: 21.
Schenk D, Basi GS and Pangalos MN. Treatment

strategies targeting amyloid β-protein.
Cold Spring
Harb. Perspect. Med
. (2012) 2: a006387.
Karl T, Garner B and Cheng D. The therapeutic

potential of the phytocannabinoid cannabidiol for

alzheimer’s disease.
Behav. Pharmacol. (2017) 28 (2
and 3-Spec Issue): 142-60.

Randy LH and Guoying B. Agonism of peroxisome

proliferator receptor-gamma may have therapeutic

potential for neuro inflammation and parkinson’s

disease.
Curr. Neuropharmacol. (2007) 5: 35–46.
Schintu N, Frau L, Ibba M, Caboni P and Garau A.

PPAR-gamma-mediated neuroprotection in a chronic

mouse model of parkinson’s disease.
Eur. J. Neurosci.
(2009) 29: 954–63.

Johri A, Calingasan NY, Hennessey TM, Sharma A and

Yang L. Pharmacologic activation of mitochondrial

biogenesis exerts widespread beneficial effects in a

transgenic mouse model of huntington’s disease.
Hum.
Mol. Genet.
(2012) 21: 1124–37.
Watson GS, Cholerton BA, Reger MA, Baker LD

and Plymate SR. Preserved cognition in patients with

early alzheimer disease and amnestic mild cognitive

impairment during treatment with rosiglitazone: a

preliminary study.
Am. J. Geriatr. Psychiatry . (2005)
13: 950-8.

Jin YN, Hwang WY, Jo C and Johnson GV. Metabolic

state determines sensitivity to cellular stress in

Huntington disease: normalization by activation of

PPARgamma.
PLoS One (2012) 7: e30406.
Jin J, Albertz J, Guo Z, Peng Q and Rudow G.

Neuroprotective effects of PPAR-γ agonist rosiglitazone

in N171-82 Q mouse model of huntington’s disease.
J.
Neurochem.
(2013) 125: 410-9.
Toledo EM and Inestrosa NC. Activation of Wnt

signaling by lithium and rosiglitazone reduced spatial

memory impairment and neurodegeneration in brains

of an APPswe/PSEN1DeltaE9 mouse model of

alzheimer’s disease.
Mol. Psychiatry. (2010) 15: 272-
85.

Liu L, Zhi Q, Shen M, Gong FR, Zhou BP Lian L,

Shen B, Chen K, Duan W, Wu MY, Tao M and Li

W. FH535, a β-catenin pathway inhibitor, represses

pancreatic cancer xenograft growth and angiogenesis.

Oncotarget
. (2016) 7: 47145-62.
Cheng A, Hou Y and Mattson MP. Mitochondria and

(26)

(27)

(28)

(29)

(30)

(31)

(32)

(33)

(34)

(35)

(36)

(37)

(38)

(39)

(40)

(41)

(42)

(43)

(44)

(45)

(46)

(47)

(48)

(49)

(50)

(51)

(52)

(53)
 
PPAR-γ involvement in Aβ neurotoxicity1417
neuroplasticity.
ASN Neuro. (2010) 2: e00045.
Silva-Alvarez C, Arrazola MS, Godoy JA, Ordenes D

and Inestrosa NC. Canonical Wnt signaling protects

hippocampal neurons from Abeta oligomers: role

of non-canonical Wnt-5a/Ca(2+) in mitochondrial

dynamics.
Front. Cell Neurosci. (2013) 7: 97.
Nimmrich V, Grimm C, Draguhn A, Barghorn S,

Lehmann
A, Schoemaker H, Hillen H, Gross G,
Ebert U and Bruehl C. Amyloid beta oligomers (A

beta(1-42) globulomer) suppress spontaneous synaptic

activity by inhibition of P/Q-type calcium currents.
J.
Neurosci.
(2008) 28: 788-97.
Yun SH, Gamkrelidze G, Stine WB, Sullivan PM

and Pasternak JF. Amyloid-beta1-42 reduces neuronal

excitability in mouse dentate gyrus.
Neurosci. Lett.
(2006) 403: 162-5.

Kaczorowski CC, Sametsky E, Shah S, Vassar R

and Disterhoft JF. Mechanisms underlying basal and

learning-related intrinsic excitability in a mouse model

of alzheimer’s disease.
Neurobiol. Aging (2011) 32:
1452-65.

Chen QS, Kagan BL, Hirakura Y and Xie CW.

Impairment of hippocampal long-term potentiation

by alzheimer amyloid beta-peptides.
J. Neurosci. Res.
(2000) 60: 65–72.

Puzzo D, Privitera L, Leznik E, Fa` M, Staniszewski

A, Palmeri A and Arancio O. Picomolar amyloid-beta

positively modulates synaptic plasticity and memory

in hippocampus.
J. Neurosci. (2008) 28: 14537–45.
Jin M, Shepardson N, Yang T, Chen G, Walsh D

and Selkoe DJ. Soluble amyloid beta-protein dimers

isolated from alzheimer cortex directly induce Tau

hyperphosphorylation and neuritic degeneration.
Proc.
Natl. Acad. Sci. USA
(2011) 108: 5819–24.
Arispe N, Rojas E and Pollard HB. Alzheimer disease

amyloid beta protein forms calcium channels in

bilayer membranes: blockade by tromethamine and

aluminium.
Proc. Natl. Acad. Sci. USA (1993) 90:
567-71.

Kawahara M and Kuroda Y. Molecular mechanism

of neurodegeneration induced by alzheimer’s beta-

amyloid protein: channel formation and disruption

of calcium homeostasis.
Brain Res. Bull. (2000) 53:
389-97.

Heneka MT, Sastre M, Dumitrescu-Ozimek L,

Hanke A and Dewachter I. Acute treatment with the

PPARgamma agonist pioglitazone and ibuprofen

reduces glial inflammation and Abeta1-42 levels in

APPV717I transgenic mice.
Brain (2005) 28 (Pt 6):
1442-53.

Feinstein DL, Galea E, Gavrilyuk V, Brosnan CF

and Whitacre CC. Peroxisome proliferator-activated

receptor-gamma agonists prevent experimental

autoimmune encephalomyelitis.
Ann. Neurol. (2001)
51: 694-702.

Patlak J. Molecular kinetics of voltage-dependent Na
+
channels. Physiol. Rev. (1991) 71: 1047–80.

Vervaeke K, Hu H, Graham LJ and Storm JF.

Contrasting effects of the persistent Na+ current on

(54)

(55)

(56)

(57)

(58)

(59)

(60)

(61)

(62)

(63)

(64)

(65)

(66)

neuronal excitability and spike timing.
Neuron (2006)
49: 257–70.

Brown JT, Chin J, Leiser SC, Pangalos MN and

Randall AD. Altered intrinsic neuronal excitability and

reduced Na+ currents in a mouse model of alzheimer’s

disease.
Neurobiol. Aging (2011) 32: 2109e1-14.
Prakash A and Kumar A. Role of nuclear receptor

on regulation of BDNF and neuroinflammation

in hippocampus of β-amyloid animal model of

alzheimer’s disease.
Neurotox. Res. (2014) 25: 335-47.
Chen TS, Lai MC, Hung TY, Lin KM, Huang CW and

Wu SN. Pioglitazone, a PPAR-γ Activator, Stimulates

BK
Ca but Suppresses IK M in Hippocampal Neurons.
Front. Pharmacol.
(2018) 9: 977.
Tubert C, Taravini IRE, Flores-Barrera E, Sánchez

GM, Prost MA, Avale ME, Tseng KY, Rela L and

Murer MG. Decrease of a current mediated by kv1.3

channels causes striatal cholinergic interneuron

hyperexcitability in experimental parkinsonism.
Cell
Rep.
(2016) 16: 2749-62.
Johnston J, Forsythe ID and Kopp-Scheinpflug C.

Going native: voltage-gated potassium channels

controlling neuronal excitability.
J. Physiol. (2010)
588 (Pt 17): 3187-200.

Golowasch J, Thomas G, Taylor AL, Patel A, Pineda

A, Khalil C and Nadim F. Membrane capacitance

measurements revisited: dependence of capacitance

value on measurement method in nonisopotential

neurons.
J. Neurophysiol. (2009) 102: 2161-75.
Gilman JP, Medalla M and Luebke JI. Area-specific

features of pyramidal neurons-a comparative study in

mouse and rhesus monkey.
Cereb. Cortex (2017) 27:
2078-94.

Matsumura R, Yamamoto H, Hayakawa T,

Katsurabayashi S, Niwano M and Hirano-Iwata A.

Dependence and homeostasis of membrane impedance

on cell morphology in cultured hippocampal neurons.

Sci. Rep.
(2018) 8: 9905.
Brager DH, Akhavan AR and Johnston D. Impaired

dendritic expression and plasticity of h-channels in the

fmr1(-/y) mouse model of fragile X syndrome.
Cell
Rep.
(2012) 1: 225-33.
Eslamizade MJ, Saffarzadeh F, Mousavi SM, Meftahi

GH, Hosseinmardi N, Mehdizadeh M and Janahmadi

M. Alterations in CA1 pyramidal neuronal intrinsic

excitability mediated by Ih channel currents in a rat

model of amyloid beta pathology.
Neurosci. (2015)
305: 279-92.

Lesage F. Pharmacology of neuronal background

potassium channels.
Neuropharmacol. (2003) 44: 1-7.
Cameron WE, Núñez-Abades PA and Kerman IA and

Hodgson TM. Role of potassium conductances in

determining input resistance of developing brain stem

motoneurons.
J. Neurophysiol. (2000) 84: 2330-9.
Pasantes-Morales H and Tuz K. Volume changes

in neurons: hyperexcitability and neuronal death.

Contrib. Nephrol.
(2006) 152: 221-40.
Yang JW, Ru J, Ma W, Gao Y, Liang Z, Liu J, Guo

JH and Li LY. BDNF promotes the growth of human

(67)

(68)

(69)

(70)

(71)

(72)

(73)

(74)

(75)

(76)

(77)

(78)

(79)

(80)
 
Bahrami F et al. / IJPR (2019), 18 (3): 1403-14181418
neurons through crosstalk with the Wnt/β-catenin

signaling pathway via GSK-3β.
Neuropeptides (2015)
54: 35-46.

Lange C, Mix E, Rateitschak K and Rolfs A. Wnt
(81)
signal pathways and neural stem cell differentiation.

Neurodegener. Dis.
(2006) 3: 76-86.