Journal Article

Upregulation of Ca 2+ -binding proteins contributes to VTA dopamine neuron survival in the early phases of Alzheimer's disease in Tg2576 mice

Giacomet, Cecilia, Melone, Marcello, Krashia, Paraskevi, Keller, Flavio, Cauzzi, Emma, Federici, Mauro, D'Amelio, Marcello, La Barbera, Livia, Conti, Fiorenzo, Viscomi, Maria Teresa, Paoletti, Ilaria, Marino, Ramona, Nobili, Annalisa, Saba, Luana, Mercuri, Nicola Biagio

2022

Overview

Recent clinical and experimental studies have highlighted the involvement of Ventral Tegmental Area (VTA) dopamine (DA) neurons for the early pathogenesis of Alzheimer's Disease (AD). We have previously described a progressive and selective degeneration of these neurons in the Tg2576 mouse model of AD, long before amyloid-beta plaque formation. The degenerative process in DA neurons is associated with an autophagy flux impairment, whose rescue can prevent neuronal loss. Impairments in autophagy can be the basis for accumulation of damaged mitochondria, leading to disturbance in calcium (Ca ) homeostasis, and to functional and structural deterioration of DA neurons. In Tg2576 mice, we performed amperometric recordings of DA levels and analysis of dopaminergic fibers in the Nucleus Accumbens - a major component of the ventral striatum precociously affected in AD patients - together with retrograde tracing, to identify the most vulnerable DA neuron subpopulations in the VTA. Then, we focused on these neurons to analyze mitochondrial integrity and Apoptosis-inducing factor (AIF) localization by electron and confocal microscopy, respectively. Stereological cell count was also used to evaluate degeneration of DA neuron subpopulations containing the Ca -binding proteins Calbindin-D28K and Calretinin. The expression levels for these proteins were analyzed by western blot and confocal microscopy. Lastly, using electrophysiology and microfluorometry we analyzed VTA DA neuron intrinsic properties and cytosolic free Ca levels. We found a progressive degeneration of mesolimbic DA neurons projecting to the ventral striatum, located in the paranigral nucleus and parabrachial pigmented subnucleus of the VTA. At the onset of degeneration (3 months of age), the vulnerable DA neurons in the Tg2576 accumulate damaged mitochondria, while AIF translocates from the mitochondria to the nucleus. Although we describe an age-dependent loss of the DA neurons expressing Calbindin-D28K or Calretinin, we observed that the remaining cells upregulate the levels of Ca -binding proteins, and the free cytosolic levels of Ca in these neurons are significantly decreased. Coherently, TUNEL-stained Tg2576 DA neurons express lower levels of Calbindin-D28K when compared with non-apoptotic cells. Overall, our results suggest that the overexpression of Ca -binding proteins in VTA DA neurons might be an attempt of cells to survive by increasing their ability to buffer free Ca . Exploring strategies to overexpress Ca -binding proteins could be fundamental to reduce neuronal suffering and improve cognitive and non-cognitive functions in AD.

• Subject: Alzheimer Disease - metabolism / Animals / Calbindin 1 - metabolism / Calbindin 2 - metabolism / Carrier Proteins - metabolism / Dopamine - metabolism / Dopaminergic Neurons - metabolism / Mice / Up-Regulation / Ventral Tegmental Area - metabolism / Ventral Tegmental Area - pathology