Award details

Four dimensional imaging in living cells

Reference	REI20568
Principal Investigator / Supervisor	Dr Clive Price
Co-Investigators / Co-Supervisors	Professor Alistair Hetherington, Dr Martin Mcainsh, Dr Paul McKean, Dr Alan Shirras
Institution	Lancaster University
Department	Biological Sciences
Funding type	Research
Value (£)	109,367
Status	Completed
Type	Research Grant
Start date	20/10/2003
End date	19/10/2004
Duration	12 months

Abstract

The equipment will support a broad base of research that includes plant cell signalling (Hetherington, McAinsh and Roberts laboratories), yeast cell biology (Price), Drosophila neurobiology (Shirras) and parasitology (McKean). The department possesses extensive imaging facilities however these are lacking in one important aspect which is the capability to carry out low light fluorescent imaging in real time. The proposed plant cell signalling research involves the analysis of transient phenomena required for the intracellular signalling of external environmental and developmental cues. Analysis of the specificity of what is a complex array of MAPK signallig pathways in plant cells requires the study of protein/protein interactions between individual components of these pathways (Roberts). This can only be achieved by the use of FRET technology in living cells. It is further proposed to analyse signal induced calcium fluxes in stomatal guard cells (Hetherington and McAinsh). Guard cell calcium signalling regulates the opening and closing of the stomata which govern gas exchange and between the plant and the environment, which has important consequences for plant growth, development and yield in crop species. To fully understand these processes in plant cells it is necessary to exploit the rich genetic resources and technology available in Arabidopsis. However the more traditional techniques of microinjection of calcium sensitive fluorescent dyes are not applicable to the analysis of calcium transients in single Arabidopsis guard cells. Again FRET based technology is the only currently available approach to the study of calcium flux in stomatal guard cells. Further work centres on the study of the cytoskeleton in yeast and Trypanosomes (Price and McKean). The cytoskeleton is composed of dynamic intracellular structures the study of which demands real time fluorescent imaging. In both of these experimetnal systems problems arise from their inherent small cell size. This places a premium on high sensitivity of detection and fluorescent light under low intensity excitation, thus avoiding the problems of photodamage to the cell and photobleaching of the fluorophore. Finally it is proposed to study the intracellular behaviour of neuroactive peptides and peptide-processing enzymes in Drosophila (Shirras). Thie project demands the detection of low intensity fluorescent light emission emanating from small, intracellular vesicles. The requested equipment satisfies the technical demands of all of the work proposed above. All of which commands substantive current BBSRC funding and which will be further underpinned by the equipment requested.

Summary

unavailable

Committee	Closed Committee - Biochemistry & Cell Biology (BCB)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	Research Equipment Initiative 2003 (RE2) [2003]
Funding Scheme	X – not Funded via a specific Funding Scheme

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