Domenicale, ChiaraMercatelli, DanielaAlbanese, FedericaNovello, SalvatoreVincenzi, FabrizioVarani, KatiaMorari, Michele2022-05-092022-05-092022-05-092022-04-0110.3390/biomedicines10040881https://infoscience.epfl.ch/handle/20.500.14299/187677WOS:000785310900001The G2019S mutation in leucine rich-repeat kinase 2 (LRRK2) is a major cause of familial Parkinson's disease. We previously reported that G2019S knock-in mice manifest dopamine transporter dysfunction and phosphoSerine129 alpha-synuclein (pSer129 alpha-syn) immunoreactivity elevation at 12 months of age, which might represent pathological events leading to neuronal degeneration. Here, the time-dependence of these changes was monitored in the striatum of 6, 9, 12, 18 and 23-month-old G2019S KI mice and wild-type controls using DA uptake assay, Western analysis and immunohistochemistry. Western analysis showed elevation of membrane dopamine transporter (DAT) levels at 9 and 12 months of age, along with a reduction of vesicular monoamine transporter 2 (VMAT2) levels at 12 months. DAT uptake was abnormally elevated from 9 to up to 18 months. DAT and VMAT2 level changes were specific to the G2019S mutation since they were not observed in LRRK2 kinase-dead or knock-out mice. Nonetheless, dysfunctional DAT uptake was not normalized by acute pharmacological inhibition of LRRK2 kinase activity with MLi-2. Immunoblot analysis showed elevation of pSer129 alpha-syn levels in the striatum of 12-month-old G2019S KI mice, which, however, was not confirmed by immunohistochemical analysis. Instead, total alpha-syn immunoreactivity was found elevated in the striatum of 23-month-old LRRK2 knock-out mice. These data indicate mild changes in DA transporters and alpha-syn metabolism in the striatum of 12-month-old G2019S KI mice whose pathological relevance remains to be established.Biochemistry & Molecular BiologyMedicine, Research & ExperimentalPharmacology & PharmacyResearch & Experimental Medicinedatg2019s knock-in[h-3]-da uptaked1994s kinase-deadlrrk2mli-2pser129 alpha-synucleinparkinson's diseasevmat2kinase 2 lrrk2parkinsons-diseasecytosolic dopamineoxidative stressmutant lrrk2expressionmutationneurodegenerationneuronsimpairmenttext::journal::journal article::research article