Understanding of brain signalling pathways furthered with fluorescence imaging

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Scientists at Carnegie Mellon University are using confocal microscopy to study the mechanisms by which membrane trafficking controls and co-ordinates the complex signalling pathways in the brain.

Despite the fact that almost all diseases can be traced to a defect in how cells respond to signals, little is known about how signalling pathways are maintained in normal cells and the changes that occur in abnormal conditions.

Using the Andor Revolution XD confocal microscope for live cell studies, Professor Manojkumar Puthenveedu of Carnegie Mellon University has led an international team of researchers to elucidate how signalling receptors are recycled to the cell membrane. The discovery of the mechanism by which signalling receptors travel back to the surface of the cell after activation and internalisation opens up a new class of therapeutic targets.

The team used live cell confocal fluorescence microscopy to label and image receptors for adrenaline and noradrenaline and those pivotal to the regulation of heart and lung function, mood, cognition and memory, digestion, and the inflammatory response.

The researchers discovered unique domains on the endosome – they termed them Actin-Stabilised Sequence-dependent Recycling Tubule (ASSERT) domains – which trap and slow down the release of receptors. These domains could potentially be targeted by pharmaceutical agents to control diseases resulting from abnormal cell signalling.

'Confocal fluorescence microscopy has been the driver for an explosion in the study of dynamic processes in living cells, but not all systems are created equal. We have created an instrument that optimises sensitivity, acquisition speed and specimen viability,' said Mark Browne, director of systems at Andor. 'The high optical efficiency of the Revolution XD spinning disk system allows the use of reduced laser power and, therefore, lowers the risk of phototoxicity to the specimen during long term experiments while Andor's IQ software ensures precise synchronisation of illumination, detection and scanning for unparalleled image quality and acquisition rates.'

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