Cardiac sympathetic denervation in 6-OHDA-treated nonhuman primates.

Valerie Joers,Heather Simmons,Corinne Jones,Jeanette Shultz,Shahrose Rahman,Marina E Emborg,Kristine Dilley,Malú G Tansey

doi:10.1371/journal.pone.0104850

Abstract

Cardiac sympathetic neurodegeneration and dysautonomia affect patients with sporadic and familial Parkinson's disease (PD) and are currently proposed as prodromal signs of PD. We have recently developed a nonhuman primate model of cardiac dysautonomia by iv 6-hydroxydopamine (6-OHDA). Our in vivo findings included decreased cardiac uptake of a sympathetic radioligand and circulating catecholamines; here we report the postmortem characterization of the model. Ten adult rhesus monkeys (5–17 yrs old) were used in this study. Five animals received 6-OHDA (50 mg/kg iv) and five were age-matched controls. Three months post-neurotoxin the animals were euthanized; hearts and adrenal glands were processed for immunohistochemistry. Quantification of immunoreactivity (ir) of stainings was performed by an investigator blind to the treatment group using NIH ImageJ software (for cardiac bundles and adrenals, area above threshold and optical density) and MBF StereoInvestigator (for cardiac fibers, area fraction fractionator probe). Sympathetic cardiac nerve bundle analysis and fiber area density showed a significant reduction in global cardiac tyrosine hydroxylase-ir (TH; catecholaminergic marker) in 6-OHDA animals compared to controls. Quantification of protein gene protein 9.5 (pan-neuronal marker) positive cardiac fibers showed a significant deficit in 6-OHDA monkeys compared to controls and correlated with TH-ir fiber area. Semi-quantitative evaluation of human leukocyte antigen-ir (inflammatory marker) and nitrotyrosine-ir (oxidative stress marker) did not show significant changes 3 months post-neurotoxin. Cardiac nerve bundle α-synuclein-ir (presynaptic protein) was reduced (trend) in 6-OHDA treated monkeys; insoluble proteinase-K resistant α-synuclein (typical of PD pathology) was not observed. In the adrenal medulla, 6-OHDA monkeys had significantly reduced TH-ir and aminoacid decarboxylase-ir. Our results confirm that systemic 6-OHDA dosing to nonhuman primates induces cardiac sympathetic neurodegeneration and loss of catecholaminergic enzymes in the adrenal medulla, and suggests that this model can be used as a platform to evaluate disease-modifying strategies aiming to induce peripheral neuroprotection.

Highlights

Cardiac sympathetic neurodegeneration and dysautonomia affect sporadic and familial cases of Parkinson’s disease (PD) [1,2,3,4], and are currently proposed to predate PD motor syndrome [5,6,7,8,9]
The present study demonstrates that systemic 6-OHDA dosing to rhesus monkeys induces cardiac sympathetic neurodegeneration and decreases catecholaminergic enzymes in the adrenal medulla
6-OHDA-treated monkeys, like many PD patients, had decreased sympathetic cardiac innervation, the extent of loss seems to be less than the human cases

Summary

Introduction

Cardiac sympathetic neurodegeneration and dysautonomia affect sporadic and familial cases of Parkinson’s disease (PD) [1,2,3,4], and are currently proposed to predate PD motor syndrome [5,6,7,8,9]. Orthostatic hypotension, fatigue, and abnormal control of electrical heart activity are typical manifestations of these pathologies. Reduced circulating catecholamines can be present and is more commonly found in PD patients exhibiting orthostatic hypotension [10,11,12]. Neuropathological studies have confirmed cardiac nerve loss showing reduced tyrosine hydroxylase (TH) and neurofilament immunostaining in epicardial nerve bundles [14,15,16]. Lewy bodies (intracytoplasmic inclusions composed mainly of aggregated a-synuclein), which are a classical pathological feature of PD, have been described in cardiac nerve structures including cardiac plexus [17], cardiac ganglia within the interatrial groove, epicardial bundles and nerves that innervate the myocardium, as well as in the adrenal medulla [18,19]

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