Implementation of improved postoperative care decreases the mortality rate of operated mice after an abundant 6-hydroxydopamine lesion of nigrostriatal dopaminergic neurons

A mouse model of Parkinson’s disease with an abundant lesion of nigrostriatal dopaminergic neurons can be achieved by stereotactic injection of 6-hydroxydopamine into the medial forebrain bundle. However, postoperative mortality can be excessively high without intensive postoperative care. Here, we show that improvements in stereotactic operations and postoperative care result in significant benefits for both animal well-being and research efficiency. Adopting a wide combination of mostly previously described improvements resulted in a decrease of postoperative mortality from 71% to 14% and an increase in successful abundant dopaminergic lesions from 46% to 81%. The techniques adopted are described in detail. In addition, we describe a simple protocol for gradual preoperative handling which can be utilized to decrease animal stress, aggressive and aversive behaviors, and to facilitate postoperative care and other subsequent handling. We propose that the implementation of these improvements greatly decreases the risk of animal suffering and that the improvements are worth adopting in any research group utilizing abundant 6-hydroxydopamine-induced dopaminergic lesions in mice. Suggestions for further improvement are also presented

Genetic lack of histamine upregulates dopamine neurotransmission and alters rotational behavior but not levodopa-induced dyskinesia in a mouse model of Parkinson’s disease

The brain histaminergic and dopaminergic systems closely interact, and some evidence also suggests significant involvement of histamine in Parkinson’s disease (PD), where dopaminergic neurons degenerate. To further investigate histamine-dopamine interactions, particularly in the context of PD, a genetic lack of histamine and a mouse model of PD and levodopa-induced dyskinesia were here combined. Dopaminergic lesions were induced in histidine decarboxylase knockout and wildtype mice by 6-hydroxydopamine injections into the medial forebrain bundle. Post-lesion motor dysfunction was studied by measuring drug-induced rotational behavior and dyskinesia. Striatal tissue from both lesioned and naïve animals was used to investigate dopaminergic, serotonergic and histaminergic biomarkers. Histamine deficiency increased amphetamine-induced rotation but did not affect levodopa-induced dyskinesia. qPCR measurements revealed increased striatal expression of D1 and D2 receptor, DARPP-32, and H3 receptor mRNA, and synaptosomal release experiments in naïve mice indicated increased dopamine release. A lack of histamine thus causes pre- and postsynaptic upregulation of striatal dopaminergic neurotransmission which may be reflected in post-lesion motor behavior. Disturbances or manipulations of the histaminergic system may thus have significant consequences for dopaminergic neurotransmission and motor behavior in both healthy and disease conditions. The findings also represent new evidence for the complex interplay between dopamine and histamine within the nigrostriatal pathway.Peer reviewe

Effects of antidyskinetic nicotine treatment on dopamine release in dorsal and ventral striatum

The treatment of Parkinson's disease is often complicated by levodopa-induced dyskinesia (LID), and antidyskinetic treatment options are currently sparse. Nicotinic acetylcholine receptors have been suggested as potential targets for treatment of LID, as nicotinic agonists have been reported to alleviate LID in animal models. We aimed at the first independent replication of an antidyskinetic effect by nicotine using a mouse model of LID, and at investigation of its mechanisms by studying the release of [H-3]dopamine from synaptosomes prepared from the dorsal and ventral striatum. Chronic nicotine treatment in drinking water inhibited the development of LID in mice lesioned unilaterally with 6-hydroxydopamine and treated chronically with levodopa and benserazide. The antidyskinetic nicotine treatment had no effect on [H-3]dopamine release mediated by alpha 4 beta 2* nicotinic receptors, but decreased alpha 6 beta 2*-mediated [H-3]dopamine release in the lesioned dorsal striatum and the ventral striatum. In addition, nicotine treatment restored [H-3]dopamine release in the lesioned ventral striatum to intact levels. The results support a role for nicotinic receptors as drug targets for treatment of LID, and suggest that striatal presynaptic alpha 6 beta 2* receptors are important mediators of nicotine's antidyskinetic effect.Peer reviewe

Dyskinesia and brain-derived neurotrophic factor levels after long-term levodopa and nicotinic receptor agonist treatments in female mice with near-total unilateral dopaminergic denervation

BackgroundThe treatment of Parkinson's disease is often complicated by levodopa-induced dyskinesia (LID). Nicotinic acetylcholine receptor agonists can alleviate LID in animal models but may be less effective in conditions of severe dopaminergic denervation. While the mechanisms of LID remain incompletely understood, elevated corticostriatal levels of the brain-derived neurotrophic factor (BDNF) have been suggested to play a role. Here, female mice with near-total unilateral 6-hydroxydopamine-induced nigrostriatal lesions were chronically treated with levodopa, and the effects of the 7 nicotinic receptor partial agonist AZD0328 and nicotine on LID were assessed. At the end of the experiment, BDNF protein levels in the prefrontal cortex and striatum were measured.ResultsFive-day treatments with three escalating doses of AZD0328 and a 10-week treatment with nicotine failed to alleviate LID. BDNF levels in the lesioned striatum correlated positively with LID severity, but no evidence was found for a levodopa-induced elevation of corticostriatal BDNF in the lesioned hemisphere. The nicotine treatment decreased BDNF levels in the prefrontal cortex but had no effect on striatal BDNF.ConclusionsThe findings suggest that treatment of LID with nicotinic agonists may lose its effectiveness as the disease progresses, represent further evidence for a role for BDNF in LID, and expand previous knowledge on the effects of long-term nicotine treatment on BDNF.Peer reviewe

Attenuated dopaminergic neurodegeneration and motor dysfunction in hemiparkinsonian mice lacking the alpha 5 nicotinic acetylcholine receptor subunit

Preclinical studies suggest the involvement of various subtypes of nicotinic acetylcholine receptors in the pathophysiology of Parkinson's disease, a neurodegenerative disorder characterized by the death of dopaminergic neurons in the substantia nigra pars compacta (SNC). We studied for the first time the effects of alpha 5 nicotinic receptor subunit gene deletion on motor behavior and neurodegeneration in mouse models of Parkinson's disease and levodopa-induced dyskinesia. Unilateral dopaminergic lesions were induced in wild-type and alpha 5-KO mice by 6-hydroxydopamine injections into the striatum or the medial forebrain bundle. Subsequently, rotational behavior induced by dopaminergic drugs was measured. A subset of animals received chronic treatments with levodopa and nicotine to assess levodopa-induced dyskinesia and antidyskinetic effects by nicotine. SNC lesion extent was assessed with tyrosine hydroxylase immunohistochemistry and stereological cell counting. Effects of alpha 5 gene deletion on the dopaminergic system were investigated by measuring ex vivo striatal dopamine transporter function and protein expression, dopamine and metabolite tissue concentrations and dopamine receptor mRNA expression. Hemiparkinsonian alpha 5-KO mice exhibited attenuated rotational behavior after amphetamine injection and attenuated levodopa-induced dyskinesia. In the intrastriatal lesion model, dopaminergic cell loss in the medial cluster of the SNC was less severe in alpha 5-KO mice. Decreased striatal dopamine uptake in alpha 5-KO animals suggested reduced dopamine transporter function as a mechanism of attenuated neurotoxicity. Nicotine reduced dyskinesia severity in wild-type but not alpha 5-KO mice. The attenuated dopaminergic neurodegeneration and motor dysfunction observed in hemiparkinsonian alpha 5KO mice suggests potential for alpha 5 subunit-containing nicotinic receptors as a novel target in the treatment of Parkinson's disease. (C) 2018 The Authors. Published by Elsevier Ltd.Peer reviewe