Parkinson’s Paradox: Alpha-synuclein’s Selective Strike on SNc Dopamine Neurons over VTA

In synucleinopathies, including Parkinson’s disease (PD), dopamine neurons in the substantia nigra pars compacta (SNc) exhibit greater vulnerability to degeneration than those in the ventral tegmental area (VTA). While α-synuclein (αSyn) pathology is implicated in nigral dopamine neuron loss, the mechanisms by which αSyn affects neuronal activity and midbrain dopamine network connectivity prior to cell death remain unclear. This study tested the hypothesis that elevated αSyn expression induces pathophysiological changes in firing activity and disrupts network connectivity dynamics of dopamine neurons before neuronal loss. We employed two mouse models of synucleinopathy: preformed αSyn fibril (PFF) injection and AAV-mediated expression of human αSyn (hαSyn) under the control of the tyrosine hydroxylase (TH) promoter, both targeting the VTA and SNc. Four weeks post-injection, brain sections underwent histological, electrophysiological, and network analyses. Immunohistochemistry for TH, hαSyn, and phospho-Ser129 αSyn assessed αSyn expression and dopaminergic neuron alterations. Neuronal viability was evaluated using two complementary approaches: quantification of TH ⁺ or FOX3 ⁺ and TUNEL labeling. Importantly, these analyses revealed no significant changes in neuronal counts or TUNEL ⁺ cells at this time point, confirming that subsequent functional assessments captured pre-neurodegenerative, αSyn-induced alterations rather than late-stage neurodegeneration. Electrophysiological recordings revealed a differential effect of hαSyn expression. SNc dopamine neurons exhibited significantly increased baseline firing rates, whereas VTA dopamine neurons remained unchanged. These findings indicate a region-specific vulnerability to αSyn-induced hyperactivity of dopamine neurons. Further analysis revealed impaired homeostatic firing rate regulation in SNc, but not VTA, dopamine neurons, demonstrated by a reduced capacity to recover baseline firing following hyperpolarization. Collectively, our results demonstrate that, prior to neurodegeneration, elevated αSyn expression differentially disrupts both basal firing activity and network stability of SNc dopamine neurons, while sparing VTA dopamine neurons. By identifying neurophysiological changes preceding dopaminergic neuron loss, these findings provide critical insights into the pathophysiological mechanisms predisposing SNc neurons to degeneration in Parkinson’s disease.