Pirouetting pigs: A large non-primate animal model based on unilateral 6-hydroxydopamine lesioning of the nigrostriatal pathway

Introduction: The rotating 6-hydroxydopamine (6-OHDA) rat model has long been important when developing new treatment strategies for Parkinson's disease (PD). Similar non-human primate models have been developed for translational research purposes as large animal models are required by regulatory bodies as an intermediate "phase 0" trial step. However, experimental research in non-human primates encounters several economical and regulatory issues, which may be avoided by the alternative use of pigs as a large animal model for experimental brain research. Objective: The primary aim of this study was to examine if unilateral injections of 6-OHDA into the Gottingen minipig nigrostriatal pathway would lead to dopaminergic imbalance and rotational behavior similar to the 6-OHDA unilateral symptomatic model of PD created in other species. The secondary aim was to attempt to verify the rotational behavior as a parkinsonian symptom using subthalamic deep brain stimulation (STN-DBS) to minimize the elicited rotational pattern. Materials and methods: Using an MRI-based stereotactic procedure, ten female Gottingen minipigs were injected unilaterally with 6-OHDA in the nigrostriatal pathway. Postoperatively, an MRI was performed, and the animals were injected with amphetamine and apomorphine and observed for rotational behavior. After a survival period of three months the brains were removed and immunohistochemically stained for tyrosine hydroxylase (TH). One week before sacrifice two animals had DBS electrodes unilaterally implanted in the subthalamic nucleus and various stimulation protocols were conducted during amphetamine challenge. Results: As expected most animals rotated towards the side of the lesion when given amphetamine (3.5-4.0 mg/kg), whereas the predicted opposite response to apomorphine were much harder to reproduce. T1- and T2 weighted postoperative MRI could demonstrate the size and the location of the 6-OHDA injection. Postmortem TH-staining of the final two animals receiving a medial and a lateral injection of 25 mu L of 6-OHDA (8 mu g/mu L, injection rate 5 mu L/min) into the diencephalic nigrostriatal pathway showed a prominent unilateral decrease in TH-staining of the substantia nigra pars compacta, the ventral tegmental area and the nigrostriatal pathway on the lesioned side. These two animals displayed spontaneous rotational behavior toward the lesioned side for the first 2-3 days postoperatively, and this behavior could later on be reelicited by amphetamine and attenuated by ipsilateral STN-DBS. Conclusion: Female Gottingen minipigs are susceptible to unilateral dopaminergic degeneration when properly injected unilaterally with sufficient amounts of 6-OHDA in the nigrostriatal pathway. The location of the 6OHDA injections and thus the accuracy of the employed stereotaxy can be verified in vivo using MRI postoperatively. The injected minipigs display unilateral parkinsonism with a well-defined rotational response to amphetamine that may be ameliated by STN-DBS performed on the lesioned side. The response to apomorphine was, however, not consistent, illustrating that further work on this promising non-primate large animal model is needed, before it is fully similar to the established 6-OHDA models in other species.