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Molecular Neurodegeneration
Published in: Molecular Neurodegeneration 1/2015

Open Access 01-12-2015 | Research article

Pharmacogenetic stimulation of cholinergic pedunculopontine neurons reverses motor deficits in a rat model of Parkinson’s disease

Authors: Ilse S. Pienaar, Sarah E. Gartside, Puneet Sharma, Vincenzo De Paola, Sabine Gretenkord, Dominic Withers, Joanna L. Elson, David T. Dexter

Published in: Molecular Neurodegeneration | Issue 1/2015

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Abstract

Background

Patients with advanced Parkinson's disease (PD) often present with axial symptoms, including postural- and gait difficulties that respond poorly to dopaminergic agents. Although deep brain stimulation (DBS) of a highly heterogeneous brain structure, the pedunculopontine nucleus (PPN), improves such symptoms, the underlying neuronal substrate responsible for the clinical benefits remains largely unknown, thus hampering optimization of DBS interventions. Choline acetyltransferase (ChAT)::Cre+ transgenic rats were sham-lesioned or rendered parkinsonian through intranigral, unihemispheric stereotaxic administration of the ubiquitin-proteasomal system inhibitor, lactacystin, combined with designer receptors exclusively activated by designer drugs (DREADD), to activate the cholinergic neurons of the nucleus tegmenti pedunculopontine (PPTg), the rat equivalent of the human PPN. We have previously shown that the lactacystin rat model accurately reflects aspects of PD, including a partial loss of PPTg cholinergic neurons, similar to what is seen in the post-mortem brains of advanced PD patients.

Results

In this manuscript, we show that transient activation of the remaining PPTg cholinergic neurons in the lactacystin rat model of PD, via peripheral administration of the cognate DREADD ligand, clozapine-N-oxide (CNO), dramatically improved motor symptoms, as was assessed by behavioral tests that measured postural instability, gait, sensorimotor integration, forelimb akinesia and general motor activity. In vivo electrophysiological recordings revealed increased spiking activity of PPTg putative cholinergic neurons during CNO-induced activation. c-Fos expression in DREADD overexpressed ChAT-immunopositive (ChAT+) neurons of the PPTg was also increased by CNO administration, consistent with upregulated neuronal activation in this defined neuronal population.

Conclusions

Overall, these findings provide evidence that functional modulation of PPN cholinergic neurons alleviates parkinsonian motor symptoms.
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Literature
1.
Winn P, Brown VJ, Inglis WL. On the relationship between the striatum and the pedunculopontine tegmental nucleus. Crit Rev Neurobiol. 1997;11:241–61.CrossRefPubMed
2.
Alessandro S, Ceravolo R, Brusa L, Pierantozzi M, Costa A, Galati S, et al. Non-motor functions in parkinsonian patients implanted in the pedunculopontine nucleus: focus on sleep and cognitive do mains. J Neurol Sci. 2010;289:44–8.CrossRefPubMed
3.
Martinez-Gonzalez C, Bolam JP, Mena-Segovia J. Topographical organization of the pedunculopontine nucleus. Front Neuroanat. 2011;5:22.PubMedCentralPubMed
4.
Hirsch EC, Graybiel AM, Duyckaerts C, Javoy-Agid F. Neuronal loss in the pedunculopontine tegmental nucleus in Parkinson disease and in progressive supranucleur palsy. Proc Natl Acad Sci U S A. 1987;84:5976–80.PubMedCentralCrossRefPubMed
5.
Pienaar IS, Elson JL, Racca C, Nelson G, Turnbull DM, Morris CM. Mitochondrial abnormality associates with type-specific neuronal loss and cell morphology changes in the pedunculopontine nucleus in Parkinson disease. Am J Pathol. 2013;183:1826–40.PubMedCentralCrossRefPubMed
6.
Zweig RM, Whitehouse PJ, Casanova MF, Walker LC, Jankel WR, Price DL. Loss of pedunculopontine neurons in progressive supranuclear palsy. Ann Neurol. 1987;22:18e25.CrossRef
7.
MacLaren DA, Santini JA, Russell AL, Markovic T, Clark SD. Deficits in motor performance after pedunculopontine lesions in rats – impairment depends on demands of task. Eur J Neurosci. 2014;40:3224–36.CrossRefPubMed
8.
Karachi C, Grabli D, Bernard FA, Tande D, Wattiez N, Belaid H, et al. Cholinergic mesencephalic neurons are involved in gait and postural disorders in Parkinson disease. J Clin Invest. 2010;120:2745–54.PubMedCentralCrossRefPubMed
9.
Clark SD, Alderson HL, Winn P, Latimer MP, Nothacker HP, Civelli O. Fusion of diphtheria toxin and urotensin II produces a neurotoxin selective for cholinergic neurons in the rat mesopontine tegmentum. J Neurochem. 2007;102:112–20.CrossRefPubMed
10.
Pienaar IS, Lu B, Schallert T. Closing the gap between clinic and cage: sensori-motor and cognitive behavioural testing regimens in neurotoxin-induced animal models of Parkinson’s disease. Neurosci Biobehav Rev. 2012;36:2305–24.CrossRefPubMed
11.
Pienaar IS, Harrison IF, Elson JL, Bury A, Woll P, Simon AK, et al. An animal model mimicking pedunculopontine nucleus cholinergic degeneration in Parkinson's disease. Brain Struct Funct. 2015;220:479–500.CrossRefPubMed
12.
Paxinos G, Watson C. The rat brain in stereotaxic coordinates. 6th ed. Sydney: Academic; 2009.
13.
Elson JL, Yates A, Pienaar IS. Pedunculopontine cell loss and protein aggregation direct microglia activation in parkinsonian rats. Brain Struct Funct. 2015; doi:10.​1007/​s00429-015-1045-4.
14.
Pienaar IS, Van de Berg W. A non-cholinergic neuronal loss in the pedunculopontine nucleus of toxin-evoked parkinsonian rats. Exp Neurol. 2013;248:213–23.CrossRefPubMed
15.
Plaha P, Gill SS. Bilateral deep brain stimulation of the pedunculopontine nucleus for Parkinson’s disease. Neuroreport. 2005;16:1883–7.CrossRefPubMed
16.
Stefani A, Lozano AM, Peppe A, Stanzione P, Galati S, Tropepi D, et al. Bilateral deep brain stimulation of the pedunculopontine and subthalamic nuclei in severe Parkinson's disease. Brain. 2007;130:1596–607.CrossRefPubMed
17.
Wilcox RA, Cole MH, Wong D, Coyne T, Silburn P, Kerr G. Pedunculopontine nucleus deep brain stimulation produces sustained improvement in primary progressive freezing of gait. J Neurol Neurosurg Psychiatry. 2011;82:1256–9.CrossRefPubMed
18.
Moro E, Hamani C, Poon YY, Al-Khairallah T, Dostrovsky JO, Hutchison WD, et al. Unilateral pedunculopontine stimulation improves falls in Parkinson’s disease. Brain. 2010;133:215–24.CrossRefPubMed
19.
Welter ML, Demain A, Ewenczyk C, Czernecki V, Lau B, El Helou A, et al. PPNa-DBS for gait and balance disorders in Parkinson’s disease: a double-blind, randomised study. J Neurol. 2015; doi:10.​1007/​s00415-015-7744-1.
20.
Jenkinson N, Nandi D, Miall RC, Stein JF, Aziz TZ. Pedunculopontine nucleus stimulation improves akinesia in a parkinsonian monkey. Neuroreport. 2004;15:2621–4.CrossRefPubMed
21.
Jenkinson N, Nandi D, Oram R, Stein JF, Aziz TZ. Pedunculopontine nucleus electrical stimulation alleviates akinesia independently of dopaminergic mechanisms. Neuroreport. 2006;17:639–41.CrossRefPubMed
22.
Mesulam MM, Mufson EJ, Wainer BH, Levey AI. Central cholinergic pathways in the rat: an overview based on alternative nomenclature (Hh1-Ch6). Neuroscience. 1983;10:1185–201.CrossRefPubMed
23.
Mena-Segovia J, Micklem BR, Nair-Roberts RG, Ungless MA, Bolam JP. GABAergic neuron distribution in the pedunculopontine nucleus defines functional subterritories. J Comp Neurol. 2009;515:397–408.CrossRefPubMed
24.
Wang HL, Morales M. Pedunculopontine and laterodorsal tegmental nuclei contain distinct populations of cholinergic, glutamatergic and GABAergic neurons in the rat. Eur J Neurosci. 2009;29:340–58.CrossRefPubMed
25.
Sharma P, Pienaar IS. Pharmacogenetic and optical dissection for mechanistic understanding of Parkinson’s disease: potential utilities revealed through behavioural assessment. Neurosci Biobehav Rev. 2014;47C:87–100.CrossRef
26.
Armbruster BN, Li X, Pausch MH, Herlitze S, Roth BL. Evolving the lock to fit the key to create a family of G protein-coupled receptors potently activated by an inert ligand. Proc Natl Acad Sci U S A. 2007;104:5163–8.PubMedCentralCrossRefPubMed
27.
Alexander GM, Rogan SC, Abbas AI, Armbruster BN, Pei Y, Allen JA, et al. Remote control of neuronal activity in transgenic mice expressing evolved G protein–coupled receptors. Neuron. 2009;63:27–39.PubMedCentralCrossRefPubMed
28.
Witten IB, Steinberg EE, Lee SY, Davidson TJ, Zalocusky KA, Brodsky M, et al. Recombinase-driver rat lines: tools, techniques, and optogenetic application to dopamine-mediated reinforcement. Neuron. 2011;72:721–33.PubMedCentralCrossRefPubMed
29.
Rye DB, Saper CB, Lee HJ, Wainer BH. Pedunculopontine tegmental nucleus of the rat: cytoarchitecture, cytochemistry, and some extrapyramidal connections of the mesopontine tegmentum. J Comp Neurol. 1987;259:483–528.CrossRefPubMed
30.
Ichinohe N, Teng B, Kitai ST. Morphological study of the tegmental pedunculopontine nucleus, substantia nigra and subthalamic nucleus, and their interconnections in rat organotypic culture. Anat Embryol (Berl). 2000;201:435–53.CrossRef
31.
Michaelides M, Anderson SA, Ananth M, Smirnov D, Thanos PK, Neumaier JF, et al. Whole-brain circuit dissection in free-moving animals reveals cell-specific mesocorticolimbic networks. J Clin Invest. 2013;123:5342–50.PubMedCentralCrossRefPubMed
32.
Morgan JI, Curran T. Stimulus-transcription coupling in the nervous system: involvement of the inducible porto-oncogenes fos and jun. Annu Rev Neurosci. 1991;14:421–51.CrossRefPubMed
33.
Ferguson SM, Phillips PE, Roth BL, Wess J, Neumaier JF. Direct-pathway striatal neurons regulate the retention of decision-making strategies. J Neurosci. 2013;33:11668–76.PubMedCentralCrossRefPubMed
34.
Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher DS, et al. Rapid, reversible activation of AgRP neurons drives feeding behavior in mice. J Clin Invest. 2011;121:1424–8.PubMedCentralCrossRefPubMed
35.
Sasaki K, Suzuki M, Mieda M, Tsujino N, Roth B, Sakurai T. Pharmacogenetic modulation of orexin neurons alters sleep/wakefulness states in mice. PLoS One. 2011;6, e20360.PubMedCentralCrossRefPubMed
36.
Mena-Segovia J, Sims HM, Magill PJ, Bolam JP. Cholinergic brainstem neurons modulate cortical gamma activity during slow oscillations. J Physiol. 2008;586:2947–60.PubMedCentralCrossRefPubMed
37.
Kang Y, Kitai ST. Electrophysiological properties of pedunculopontine neurons and their postsynaptic responses following stimulation of substantia nigra reticulata. Brain Res. 1990;535:79–95.CrossRefPubMed
38.
Leonard CS, Llinás RR. Electrophysiology of mammalian pedunculopontine and laterodorsal tegmental neurons in vitro: implications for the control of REM sleep. In: Steriade M, Biesold D, editors. Brain cholinergic systems. New York: Oxford University Press; 1990. p. 205–23.
39.
Takakusaki K, Kitai ST. Ionic mechanisms involved in the spontaneous firing of the tegmental pedunculopontine nucleus (PPN) neurons of the rat. Neuroscience. 1997;78:771–94.CrossRefPubMed
40.
Kim J, Nakajima K, Oomura Y, Wayner MJ, Sasaki K. Electrophysiological effects of orexins/hypocretins on pedunculopntine tegmental neurons in rats: an in vitro study. Peptides. 2009;30:191–209.CrossRefPubMed
41.
Vincent SR, Satoh K, Armstrong DM, Fibiger HC. NADPH diaphorase: a selective histochemical marker for the cholinergic neurons of the pontine reticular formation. Neurosci Lett. 1983;43:31–6.CrossRefPubMed
42.
Takakusaki K, Shiroyama T, Yamamoto T, Kitai ST. Cholinergic and noncholinergic tegmental pedunculopontine projection neurons in rats revealed by intracellular labelling. J Comp Neurol. 1996;371:345–61.CrossRefPubMed
43.
Zhang QJ, Liu J, Wang Y, Wang S, Wu ZH, Yan W, et al. The firing activity of presumed cholinergic and non-cholinergic neurons of the pedunculopontine nucleus in 6-hydroxydopamine-lesioned rats: an in vivo electrophysiological study. Brain Res. 2008;1243:152–60.CrossRefPubMed
44.
Breit S, Bouali-Benazzouz R, Benabid AL, Benazzouz A. Unilateral lesion of the nigrostriatal pathway induces an increase of neuronal activity of the pedunculopontine nucleus, which is reversed by the lesion of the subthalamic nucleus in the rat. Eur J Neurosci. 2001;14:1833–42.CrossRefPubMed
45.
Jeon MF, Ha Y, Cho YH, Lee BH, Park YG, Chang JW. Effect of ipsilateral subthalamic nucleus lesioning in a rat parkinsonian model: study of behavior correlated with neuronal activity in the pedunculopontine nucleus. J Neurosurg. 2003;99:762–7.CrossRefPubMed
46.
Lewis GN, Byblow WD. Altered sensorimotor integration in Parkinson’s disease. Brain. 2002;125:2089–99.CrossRefPubMed
47.
Timofeeva E, Dufresne C, Sik A, Zhang ZW, Deschênes M. Cholinergic modulation of vibrissal receptive fields in trigeminal nuclei. J Neurosci. 2005;25:9135–43.CrossRefPubMed
48.
Steidl S, Wang H, Wise RA. Lesions of cholinergic pedunculopontine tegmental nucleus neurons fail to affect cocaine or heroin self-administration or conditioned place preference in rats. PLoS One. 2014;9, e84412.PubMedCentralCrossRefPubMed
49.
Yarnall A, Rochester L, Burn DJ. The interplay of cholinergic function, attention, and falls in Parkinson’s disease. Mov Disord. 2011;26:2496–503.
50.
Bohnen NI, Muller ML, Koeppe RA, Studenski SA, Kilbourn MA, Frey KA, et al. History of falls in Parkinson disease is associated with reduced cholinergic activity. Neurology. 2009;73:1670–6.PubMedCentralCrossRefPubMed
51.
Rochester L, Yarnall AJ, Baker MR, David RV, Lord S, Galna B, et al. Cholinergic dysfunction contributes to gait disturbances in early Parkinson’s disease. Brain. 2012;135:2779–88.PubMedCentralCrossRefPubMed
52.
Di Lazzaro V, Oliviero A, Tonali PA, Marra C, Daniele A, Profice P, et al. Non-invasive in vivo assessment of cholinergic cortical circuits in AD using transcranial magnetic stimulation. Neurology. 2002;13:392–7.CrossRef
53.
Hammond C, Rouzaire-Dubois B, Féger J, Jackson A, Crossman AR. Anatomical and electrophysiological studies on the reciprocal projections between the subthalamic nucleus and nucleus tegmenti pedunculopontinus in the rat. Neuroscience. 1983;9:41–52.CrossRefPubMed
54.
Clarke PB, Hommer DW, Pert A, Skirboll LR. Innervation of substantia nigra neurons by cholinergic afferents from pedunculopontine nucleus in the rat: neuroanatomical and electrophysiological evidence. Neuroscience. 1987;23:1011–19.CrossRefPubMed
55.
Bevan MD, Bolam JP. Cholinergic, GABAergic, and glutamate-enriched inputs from the mesopontine tegmentum to the subthalamic nucleus in the rat. J Neurosci. 1995;15:7105–20.PubMed
56.
Dexter DT, Jenner P. Parkinson disease: from pathology to molecular disease mechanisms. Free Radic Biol Med. 2013;62:132–44.CrossRefPubMed
57.
Bergman H, Wichmann T, Karmon B, DeLong MR. The primate subthalamic nucleus. II. Neuronal activity in the MPTP model of parkinsonism. J Neurophysiol. 1994;72:507–20.PubMed
58.
Magill PJ, Bolam JP, Bevan MD. Dopamine regulates the impact of the cerebral cortex on the subthalamic nucleus–globus pallidus network. Neuroscience. 2001;106:313–30.CrossRefPubMed
59.
Woodlee MT, Asseo-Garcia AM, Zhao X, Liu SJ, Jones TA, Schallert T. Testing forelimb placing “across the midline” reveals distinct, lesion-dependent patterns of recovery in rats. Exp Neurol. 2005;191:310–17.CrossRefPubMed
60.
Kirik D, Rosenblad C, Björklund A. Characterization of behavioural and neurodegenerative changes following partial lesions of the nigrostriatal dopamine system induced by intrastriatal 6-hydroxydopamine in the rat. Exp Neurol. 1998;152:259–77.CrossRefPubMed
61.
Lundblad M, Andersson M, Winkler C, Kirik D, Wierup N, Cenci MA. Pharmacological validation of behavioural measures of akinesia and dyskinesia in a rat model of Parkinson’s disease. Eur J Neurosci. 2002;15:120–32.CrossRefPubMed
62.
Grabli D, Karachi C, Folgoas E, Monfort M, Tande D, Clark S, et al. Gait disorders in parkinsonian monkeys with pedunculopontine nucleus lesions: a tale of two systems. J Neurosci. 2013;33:11986–93.CrossRefPubMed
63.
Nakagawa M, Ohgod M, Nishizawa Y, Ogura H. Dopaminergic agonists and muscarinic antagonists improve lateralization in hemiparkinsonian rats in a novel exploratory Y-maze. J Pharmacol Exp Ther. 2004;309:737–44.CrossRefPubMed
64.
Pienaar IS, Lee CH, Elson JL, McGuinness L, Gentleman SM, Kalaria RN, et al. Deep-brain stimulation associates with improved microvascular integrity in the subthalamic nucleus in Parkinson’s disease. Neurobiol Dis. 2015;74C:392–405.CrossRef
65.
Root DH, Mejias-Aponte CA, Qi J, Morales M. Role of glutamatergic projections from ventral tegmental area to lateral habenula in aversive conditioning. J Neurosci. 2014;34:13906–10.PubMedCentralCrossRefPubMed
66.
67.
Oorschot DE. Total number of neurons in the neostriatal, pallidal, subthalamic, and substantia nigral nuclei of the rat basal ganglia: a stereological study using the cavalieri and optical dissector methods. J Compar Neurol. 1996;366:580–99.CrossRef
68.
Maskos U. The cholinergic mesopontine tegmentum is a relatively neglected nicotinic master modulator of the dopaminergic system: relevance to drugs of abuse and pathology. Br J Pharmacol. 2008;153:S438–45.PubMedCentralCrossRefPubMed
69.
70.
West MJ, Gundersen HJ. Unbiased stereological estimation of the number of neurons in the human hippocampus. J Comp Neurol. 1990;296:1–22.CrossRefPubMed
Metadata
Title
Pharmacogenetic stimulation of cholinergic pedunculopontine neurons reverses motor deficits in a rat model of Parkinson’s disease
Authors
Ilse S. Pienaar
Sarah E. Gartside
Puneet Sharma
Vincenzo De Paola
Sabine Gretenkord
Dominic Withers
Joanna L. Elson
David T. Dexter
Publication date
01-12-2015
Publisher
BioMed Central
Published in
Molecular Neurodegeneration / Issue 1/2015
Electronic ISSN: 1750-1326
DOI
https://doi.org/10.1186/s13024-015-0044-5

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