Objective/Rationale:
Mutations in Leucine Rich Repeat Kinase 2 (LRRK2) are implicated in Parkinson’s disease in humans. At present we understand neither the normal nor the pathogenic role of LRRK2. We need better understanding so we can design drug therapies. Antibodies are research tools that help us to map the location of proteins and discover their interactions with other proteins. MJFF has developed new antibodies for LRRK2 and this proposal will determine which experimental conditions work best to maximize these “tools” to their full scientific potential.
Project Description:
Antibodies will be tested on thin brain tissue sections mounted onto microscope slides. A color signal is generated via an immunological detection system. Brain tissue will include (a) normal pre-clinical models (b) pre-clinical models that have their LRRK2 gene taken away genetically (“knockout”) and (c) pre-clinical models that have been genetically altered to express the human LRRK2 protein, as well as their own LRRK2. The knockout is an important negative control because the antibodies should not produce any signal in this tissue. The pre-clinical models with the human LRRK2 protein serve as an additional control because we already know they express higher levels of LRRK2, thus we expect a much stronger signal in these animals. By comparing the three groups with different experimental conditions (tissue processing, buffers, detection kits) we hope to maximize the “signal to noise” ratio and obtain a clear idea of which brain cells LRRK2 is present in.
Relevance to Diagnosis/Treatment of Parkinson’s Disease:
Understanding which cells express LRRK2 gives us clues to which neurotransmitter systems/brain circuitries rely on normal LRRK2 function. Focusing more intensive research on these areas in LRRK2 disease models (e.g. transgenic pre-clinical models) may unveil specific changes that happen in disease. These changes may serve as “biomarkers” which can aid diagnosis and also may allow researchers to narrow down particular pathways that ultimately can be targeted by drugs.
Anticipated Outcome:
This work will provide optimal /standardized protocols that can be applied by labs developing pre-clinical models of LRRK2 Parkinson’s disease to map protein expression, to perform co-localization studies with other proteins of interest and to detect LRRK2 expression level or localization changes in other Parkinson’s (e.g. genetic and toxin) models.
This study is a part of MJFF’s LRRK2 Antibody Comparison Effort 2011.