Adenosine A2A receptor antagonists exert motor and neuroprotective effects by distinct cellular mechanisms.

OBJECTIVE: To investigate whether the motor and neuroprotective effects of adenosine A(2A) receptor (A(2A)R) antagonists are mediated by distinct cell types in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) model of Parkinson's disease. METHODS: We used the forebrain A(2A)R knock-out mice coupled with flow cytometric analyses and intracerebroventricular injection to determine the contribution of A(2A)Rs in forebrain neurons and glial cells to A(2A)R antagonist-mediated motor and neuroprotective effects. RESULTS: The selective deletion of A(2A)Rs in forebrain neurons abolished the motor stimulant effects of the A(2A)R antagonist KW-6002 but did not affect acute MPTP neurotoxicity. Intracerebroventricular administration of KW-6002 into forebrain A(2A)R knock-out mice reinstated protection against acute MPTP-induced dopaminergic neurotoxicity and attenuated MPTP-induced striatal microglial and astroglial activation. INTERPRETATION: A(2A)R activity in forebrain neurons is critical to the control of motor activity, whereas brain cells other than forebrain neurons (likely glial cells) are important components for protection against acute MPTP toxicity.

Uncovering a link between a plastid translocon component and rhomboid proteases using yeast mitochondria-based assays.

Rhomboid proteases are present in bacteria, insects, yeasts, parasites, mammals and plants. These proteases are part of the regulated intramembrane proteolysis mechanism for controlling processes such as development, stress response, lipid metabolism and mitochondrial membrane remodeling. Specific rhomboid protease substrates linked to these processes have been identified from insects to mammals, but not for plants. Identification of a link is a key step for elucidating the role of each rhomboid protease. Here, using a yeast mitochondria-based approach, we report evidence of a potential link between a plastid translocon component and organellar rhomboid proteases. This identification expands the types of processes involving regulated intramembrane proteolysis potentially to include at least one aspect of plastid protein transport.

Evidence that the plastid translocon Tic40 components possess modulating capabilities.

The transport of proteins into the plastid is a process that faces changing cellular needs such as the situation found in different plant organs or developing tissues. The plastid translocon must therefore be responsive to the changing cell environment to deliver efficiently different arrays of structurally diverse proteins. Although the Tic40-related envelope proteins appear to be translocon components designed to address the varying needs of protein translocation, details of their involvement remain elusive. This study was thus designed to combine plant-based experiments and yeast mitochondrion-based approaches for unveiling clues related to how the Tic40 components may behave during the protein translocation process. The main findings related to how Tic40 proteins may work are: 1) natural fluctuations are apparent in developing tissues, in different organs of the same plant, and in different species; 2) transgenic Arabidopsis seedlings can tolerate functionally a wide range of variations in Tic40 levels, from partial suppression to excessive production; 3) the Tic40 proteins themselves exhibit configurational changes in their association with yeast mitochondria in response to different carbon sources; 4) the presence of Tic40 proteins in yeast mitochondria influences regulatory aspects of the mitochondrial translocon; and 5) the Tic40 proteins associate with mitochondrial translocon components involved in regulatory-like events. The combined data provide evidence that Tic40 proteins possess modulating capabilities.

The Tic40 translocon components exhibit preferential interactions with different forms of the Oee1 plastid protein precursor.

The transport of proteins across the plastid envelope involves a host of proteinaceous components that attend to varying structural needs of the process. This study focuses on interactions between two select forms (designated Tic40 and Toc36) of the Tic40-related components and different structural versions of the Oee1 precursor to dissect the components' mode of operation. Interaction profiling revealed several features pertaining to how Tic40-related components might work during the transport process. The main operational features revealed are: (1) Tic40 interacts preferentially with Oee1 precursors containing only the plastid-targeting domain, (2) Toc36 interacts preferentially with Oee1 precursors containing both plastid- and thylakoid lumen-targeting domains, (3)carboxyl truncations to either the entire Oee1 precursor or Toc36 affect interactions negatively, and (4) the general reduction of Tic40-related protein levels in transgenic plants appears to exert a greater negative impact on endogenous Oee1 levels than the other proteins assessed, a trend that corroborates the findings of the protein interaction experiments.