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Proceedings Paper

Monitoring calcium concentration in dendritic spines of cultured hippocampal neurons with cameleons
Author(s): Xiuli Liu; Wei Zhou; Xiangning Li; Hui Gong; Qingming Luo
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Paper Abstract

Transient and substantial elevation of postsynaptic calcium was important for hippocampal long-term potentiation (LTP), so detection of calcium changes in spine was necessary to understand the mechanisms underlying synaptic plasticity. Unfortunately most recent calcium fluorescence indicators severely perturbed calcium transients, and traditional cameleons’ poor dynamic ranges prevented detection of changes of calcium. We presented a new method to monitor quantificationally free calcium concentration in dendritic spines with a new yellow cameleon (YC3.60) basing on culture of hippocampal neurons and calcium phosphate transfection technique and confocal microscopy with 458nm laser. In transiently transfected hippocampal neurons, the ratio of YFP to CFP was detected as FRET level. In our study, we got the parameters of YC3.60 excited with 458nm laser. Under control conditions, FRET levels in different dendritic spines of cultured hippocampal neurons were diverse but showed robust increases upon treatment with potassium chloride. FRET levels in different parts of hippocampal neurons were also different, the calcium concentration decreased with the distance from soma. These results suggested that the FRET methodology with YC3.60 could monitor calcium concentration in spines and it might be useful in analyzing mechanisms underlying synaptic plasticity.

Paper Details

Date Published: 31 March 2005
PDF: 9 pages
Proc. SPIE 5703, Plasmonics in Biology and Medicine II, (31 March 2005); doi: 10.1117/12.592075
Show Author Affiliations
Xiuli Liu, Huazhong Univ. of Science and Technology (China)
Wei Zhou, Huazhong Univ. of Science and Technology (China)
Xiangning Li, Huazhong Univ. of Science and Technology (China)
Hui Gong, Huazhong Univ. of Science and Technology (China)
Qingming Luo, Huazhong Univ. of Science and Technology (China)


Published in SPIE Proceedings Vol. 5703:
Plasmonics in Biology and Medicine II
Tuan Vo-Dinh; Joseph R. Lakowicz; Zygmunt K. Gryczynski, Editor(s)

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