Abstract
A devastating movement disorder affecting older population is Parkinson's disease (PD), characterized by the
abnormal deposition of alpha-Synuclein (a-Syn) in neuronal aggregates, and belongs to a group of neurological
disorders termed a-Synucleinopathies. Other PD hallmarks are loss of dopaminergic neurons from substantia
nigra and mitochondrial dysfunctions. A highly conserved and less understood family of human Tubulin
Polymerization Promoting Proteins (TPPP) shows aggregation promoting properties with a-Syn and forms
pathological protein deposits in a-Synucleinopathies. However, the in vivo role of mammalian TPPP is still largely
uncharacterized, thus creating a clear knowledge gap in our understanding of how TPPP aggregation and
neurotoxicity contributes to PD. Our recent studies were aimed at addressing this knowledge gap by developing
a new Drosophila model of PD utilizing a mutation in the only Drosophila homolog of human TPPP, named
Ringer, which recapitulates many of the salient features of human PD patients including progressive locomotor
disabilities, loss of dopaminergic neurons and mitochondrial dysfunctions. Our studies demonstrate that
TPPP/Ringer is a mitochondrial protein and loss of Ringer affects multiple components of the mitochondrial
machinery, such as the fission/fusion as indicated by elevated levels of Mfn1 and Opa1, decreased levels of
Drp1, and Complex I of mitochondrial respiratory chain. Furthermore, our findings showed that Ringer modulates
the level of aggregated human a-Syn in adult brain neurons. While these studies capitalized on the strengths of
Drosophila, the overall objective of this proposal is to extend findings from our Drosophila model to human PD
patient brain tissues and healthy controls and assess the extent to which the levels and/or localization of TPPP,
a-Syn and various mitochondrial proteins are altered with the goal of finding potential therapeutics that could
target the affected proteins/mechanisms. The central hypothesis of the proposed studies is that human PD
patient brains will display altered localization/levels of various mitochondrial proteins resembling Ringer mutants
and show similar co-aggregation of TPPP/a-Syn. We will use postmortem PD patient and healthy control
samples from similar brain regions of both genders to perform immunostaining/immunoblot analyses of TPPP
and a-Syn to examine any alteration in their localizations/levels. We will also determine the subcellular
localization/levels of various mitochondrial proteins that are part of the mitochondrial dynamics (fission/fusion
machinery, such as Drp1, Mfn1/2 and Opa1), electron transport chain such as Complex I proteins and members
of TOM complex in control and PD patient brains. Thus, launching from Drosophila, our studies will explore the
broader mechanistic underpinnings of TPPP/a-Syn mediated neurotoxicity as well as TPPP-associated
mitochondrial functions in neurodegeneration in PD and related disorders through comparative analyses. This
will provide further impetus for utilizing Drosophila as an in vivo discovery platform for genetic, proteomic and
pharmacological screens to uncover therapeutic targets to prevent or delay the progression of PD.