Shank3 deficiency severely dampens calcium buffer capacity in dopaminergic neurons from the ventral tegmental area
Abstract
The intracellular calcium buffer system plays a critical role in shaping the spatiotemporal profile of calcium signaling. In contrast to calcium influx and clearance from the cytosol, our understanding of the calcium buffer system and its role in neuropsychiatric disorders is limited. Shank3 deficiency is a key risk factor for autism and other psychiatric disorders, and many important downstream mechanisms, relating to signaling pathways, cellular physiology and morphology, and neuronal circuitry, have been uncovered. Here we report prominent alterations in the calcium signaling profile of Shank3-/- dopaminergic (DA) neurons in the ventral tegmental area (VTA). Calcium transients from Shank3-/- DA neurons exhibit enhanced amplitudes and faster decay rates. Subsequent proteomic analysis of VTA brain tissue revealed that parvalbumin (PV) expression was significantly downregulated, which was subsequently confirmed by immunostaining results. As PV is an important component of the endogenous calcium buffer system, we employed the added-buffer approach together with 2-photon and ratiometric calcium and demonstrated a reduction in the endogenous calcium buffer capacity (KE) in Shank3-/- neurons. Therefore, the downregulation of PV protein and the associated decrease in calcium buffer capacity are key pathogenic factors resulting from Shank3 deficiency.
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