Redistribution of DAT/α-synuclein complexes visualized by “in situ” proximity ligation assay in transgenic mice modelling early Parkinson’s disease

Alpha-synuclein, the major component of Lewy bodies, is thought to play a central role in the onset of synaptic dysfunctions in Parkinson's disease (PD). In particular, α-synuclein may affect dopaminergic neuron function as it interacts with a key protein modulating dopamine (DA) content at the synapse: the DA transporter (DAT). Indeed, recent evidence from our "in vitro" studies showed that α-synuclein aggregation decreases the expression and membrane trafficking of the DAT as the DAT is retained into α-synuclein-immunopositive inclusions. This notwithstanding, "in vivo" studies on PD animal models investigating whether DAT distribution is altered by the pathological overexpression and aggregation of α-synuclein are missing. By using the proximity ligation assay, a technique which allows the "in situ" visualization of protein-protein interactions, we studied the occurrence of alterations in the distribution of DAT/α-synuclein complexes in the SYN120 transgenic mouse model, showing insoluble α-synuclein aggregates into dopaminergic neurons of the nigrostriatal system, reduced striatal DA levels and an altered distribution of synaptic proteins in the striatum. We found that DAT/α-synuclein complexes were markedly redistributed in the striatum and substantia nigra of SYN120 mice. These alterations were accompanied by a significant increase of DAT striatal levels in transgenic animals when compared to wild type littermates. Our data indicate that, in the early pathogenesis of PD, α-synuclein acts as a fine modulator of the dopaminergic synapse by regulating the subcellular distribution of key proteins such as the DAT

A Geant4 simulation code for simulating optical photons in SPECT scintillation detectors

Geant4 is an object oriented toolkit created for the simulation of High-Energy Physics detectors. Geant4 allows an accurate modeling of radiation sources and detector devices, with easy configuration and friendly interface and at the same time with great accuracy in the simulation of physical processes. While most Monte Carlo codes do not allow the simulation of the transport and boundary characteristics for optical photons transport generated by scintillating crystal, Geant4 allows the simulation of the optical photons. In this paper we present an application of the Geant4 program for simulating optical photons in SPECT cameras. We aim to study the light transport within scintillators, photomultiplier tubes and coupling devices. To this end, we simulated a detector based on a scintillator, coupled to a photomultiplier tube through a glass window. We compared simulated results with experimental data and theoretical models, in order to verify the good matching with our simulations. We simulated a pencil beam of 140 keV photons impinging the crystal at different locations. For each condition, we calculated the value of the Pulse Height Centroid and the spread of the charge distribution, as read out by the anode array of the photomultiplier. Finally, the spatial and the energy resolutions of the camera have been estimated by simulated data. In all cases, we found that simulations agree very well with experimental data