Low dose DMSO treatment induces oligomerization and accelerates aggregation of α-synuclein

Dimethyl sulfoxide (DMSO) is a highly utilized small molecule that serves many purposes in scientific research. DMSO offers unique polar, aprotic and amphiphilic features, which makes it an ideal solvent for a wide variety of both polar and nonpolar molecules. Furthermore, DMSO is often used as a cryoprotectant in cell-based research. However, recent reports suggest that DMSO, even at low concentration, might interfere with important cellular processes, and cause macromolecular changes to proteins where a shift from alpha -helical to beta -sheet structure can be observed. To investigate how DMSO might influence current research, we assessed biochemical and cellular impacts of DMSO treatment on the structure of the aggregation-prone protein alpha -synuclein, which plays a central role in the etiology of Parkinson's disease, and other brain-related disorders, collectively termed the synucleinopathies. Here, we found that addition of DMSO increased the particle-size of alpha -synuclein, and accelerated the formation of seeding-potent fibrils in a dose-dependent manner. These fibrils made in the presence of DMSO were indistinguishable from fibrils made in pure PBS, when assessed by proteolytic digestion, cytotoxic profile and their ability to seed cellular aggregation of alpha -synuclein. Moreover, as evident through binding to the MJFR-14-6-4-2 antibody, which preferentially recognizes aggregated forms of alpha -synuclein, and a bimolecular fluorescence complementation assay, cells exposed to DMSO experienced increased aggregation of alpha -synuclein. However, no observable alpha -synuclein abnormalities nor differences in neuronal survival were detected after oral DMSO-treatment in either C57BL/6- or alpha -synuclein transgenic F28 mice. In summary, we demonstrate that low concentrations of DMSO makes alpha -synuclein susceptible to undergo aggregation both in vitro and in cells. This may affect experimental outcomes when studying alpha -synuclein in the presence of DMSO, and should call for careful consideration when such experiments are planned.